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,.;;_:;;;;;,~
United States
((l'i.j)) De~artment of
~ Agnculture
Forest Service
Rocky Mountain
Forest and Range
Experiment Station
Fort Collins,
Colorado 80526
General Technical
Report RM-224
Lichens as Bioindicators
of Air Quality
Stolte, Ken, and others.
1993. Lichens as bioindicators
of Agriculture,
Department
of air quality. Gen. Tech. Rep. RM-224. Fort Collins, CO: U.S.
Forest Service, Rocky Mountain Forest and Range Experiment
Station.
131 p.
Abstract
This report is
of
held in Denver,
Colorado on April 9-11, 1991. It summarizes the current
for using lichens to monitor air quality. Experts in lichenology and ecology contributed
on lichen floristics, characterization of monitoring sites, lichen species and communities, identifying
the result
a workshop
literature and techniques
information
lichen species sensitive
studies as examples
to pollutants,
of
active monitoring
lichen biomonitoring
with transplants, chemical
analysis
of
lichens,
and case
scenarios.
Keywords: lichens, air quality, biomonitoring, lichen floristics, lichen communities, gradient analysis, air
pollution, sensitive lichen species, elemental analysis, study designs, quality assurance/quality control.
Descriptions of front cover photos, clockwise starting in the upper left corner:
Fl.
Hypogymnia
enteromorpha,
Used in the Tongass
Biomonitoring Project,
concentrations
of
found on trees.
National Forest Air Quality
it accumulates
than other
for biomonitoring. Photo by
Sylvia Duran Sharnoff and Stephen Sharnoff.
lichens analyzed
Lecanora melanophthalma (Ram.) Ram., also
Rhizoplaca melanophthalma (Ram.) Leuk
and Poelt. This is the best pollution indicator of the
xerophytic lichens, referred to in Rope and Pearson
(1990). It is an umbilicate species usually found on
basalt. It is easily pried loose from the substrate; its
as
prominent
and distinctive
apothecia
make
It is very sensitive to
gaseous pollutants such as sulfur dioxide and also
accumulates heavy metals and other elements. This
identification
easy and certain.
photo shows the habitat
number
muralis,
A
L. melanophthalma.
other species are also pictured, including
christoi, calcarea, Lecidea tessilata,
Candelariella rosulans,
Acarospora americana,
Lorenz Pearson.
F3.
of
of
Caloplaca saxicola,
and others. Photo by
Usnea hirta (L.) Weber ex Wigg. is a
fruticose lichen in western conifer forests,
growing
mostly on partly sunny branches
F4.
and trunks
After five years of operation of a coal-fired
power plant at Colstrip, Montana, U. hirta thalli
within 15 km of the plant had significantly higher
sulfur content than thalli at distances of more than 40
km, though no differences in physiology or
appearance were noted. The yellowish foliose
species is Parmeliopsis ambiqua, not known to be
of
trees.
particularly
sensitive to pollutants
mostly at the bases
of
because it lives
trees. Photo by
Sharon Eversman.
Xanthoria polycarpa (Ehrh.) Olive. on
This species is tolerant to relatively
However,
heavy levels of atmospheric pollution.
even though it survives in city environments and
F5.
sagebrush bark.
A lector ia sarmentosa
lichen on the Tongass
Coast Indians
common
F2.
known
goats and was used
for bandages and baby
diapers. Alectoria sarmentosa was collected for
tissue analysis by Tongass Air Quality Biomonitoring
Project. Photo by Sylvia Duran Sharnoff and
Stephen Sharnoff.
by Northwest
higher
most ambient pollutants
deer and mountain
black-tailed
is the most common
National
Forest and the only
Alectoria in SE Alaska that drapes from trees.
provides important winter survival forage for
It
other polluted
pollution
areas, it is less abundant where
is heavy.
Photo by Lorenz Pearson.
Back-cover photos described on the inside back cover.
USDA Forest Service
General Technical
Report RM-224
Lichens as Bioindicators
of Air Quality
Technical
Coordinators:
Ken Stolte, USDA-Forest Service, Southeast Forest Experiment Station
Deborah Mangis, Environmental Protection Agency, AREAL
Robert Doty, USDA-Forest Service, Pacific Southwest Region
Kathy Tonnessen, Air Quality Division, National Park Service
Editor:
Laurie Stroh Huckaby
Rocky Mountain Forest and Range Experiment Station
Rocky Mountain Forest and Range
Experiment Station
Forest Service
U.S. Department of Agriculture
Fort Collins, Colorado
March 1993
Preface
We are pleased to offer this document
summarizing
conference
the
recommendations
participants
of
a group
the
of
on issues relating to air
pollution and lichens. These recommendations were
compiled at a workshop held in Denver, Colorado, on
April 9-11, 1991. The workshop was organized and
co-sponsored by the Washington Office, Watershed
and Air Staff of the U.S. Forest Service and the Air
Quality Division of the National Park Service. This
workshop brought together many of the experts in
John Christiano
Chief,
Air
National
Quality
field of lichenology
our understanding
of
lichens, lichen inventory
and the range
of
uses
appreciate the efforts
Park Service
air pollution on
methods,
of lichens as biomonitors.
of the organizers and
to this document
contributors
of
and monitoring
We
and wish to especially
thank the U.S. Forest Service,
Rocky Mountain
Forest and Range Experiment
Station, whose staff
graciously
agreed to edit and publish
Donna Lamb
Division
to discuss ways to improve
the effects
Air
Program Manager
USDA-Forest Service
this document.
Contents
1.
Introduction
1
Ken Stolte, Deborah Mangis, Robert Doty
2.
Floristics
6
Thomas Nash ID, Clifford Wetmore, William Anderson,
Charis Bratt, William Denison, Sharon Eversman, Barbara Murray,
Larry
St.
Clair
3. Site Characterization
16
Ken Stolte, Sherry Pittam, Roger Rosentreter
4.
Species and Communities
Cliff Smith, Linda Geiser,
41
Larry Gough, Brace McCune,
Brace Ryan, Ray Showman
5. Identification
of Sensitive Species
67
Jayne Belnap, Lorene Sigal, William Moir, Sharon Eversman
6. Active
Monitoring
Lorentz
7.
89
Pearson
Collection and Chemical Analysis of Lichens for Biomonitoring
96
Larry Jackson, Jesse Ford, David Schwartzman
8.
Lessons from Seven Case Studies
116
Ken Stolte, Robert Doty, Kathy Tonnessen, Rich Fisher
Appendix
I
Appendix
II
-
List of Participants
- List
of Reviewers
128
130
Introduction
Ken Stolte, Deborah Mangis, Robert Doty
THE WORKSHOP
due to their ability
to concentrate
elements that may
in low concentrations in the ambient air
(Tyler 1989; Nieboer et al. 1972).
be present
Park Service, Air Quality Division
The National
(NPS-AQD)
USDA
and the
convened a three-day
Forest Service (USFS)
in April 1991,
workshop
Lichens
are
relatively
easy to collect,
analyze for pollutant-related
preserve, and
studies. Because
can convert atmospheric
experts, and researchers who specialize
other terrestrial and aquatic plants, lichens
of lichens (see Appendix
in the study
nitrogen to
a
attended by federal land managers, regulatory
they
form usable by
are
cycling in some types of ecosystems (McCune 1992).
number of federal land
For these reasons,
(2) to review existing methods and
recommend standard research and monitoring
National
procedures to study lichen populations;
and the Bureau of Land Management,
contributors
to nutrient
arena.
Park Service, the Fish and Wildlife
had an ongoing
interest
studying
the
Service,
along with
regulatory agencies and the regulated industries
in
and (3) to
of data on lichen responses to
air pollution within the regulatory
to be important
management agencies such as the Forest Service,
quality;
assess the usefulness
considered
a
is
I).
The objectives of the
known about
workshop were: (1) to review what
the suitability of lichens as bioindicators
of air
lichens
have
to
of air pollutants
(Ryan and Rhoades 1991;
Rhoades 1988; Wetmore 1983; Gough and Erdman
determine the presence and effects
on natural ecosystems
RATIONALE
1971;
a
are an
stomata
lack the control
over gas exchange as vascular plants do (Larcher
1975).
Lichens were first recognized
sensitive to high concentrations
as organisms
of gaseous pollutants
such as sulfur dioxide (Skye 1968; Rose and
Hawksworth 1981). For this reason they have been
used as indicators of urban pollution and point-source
emissions
from uncontrolled
combustion
sources.
They were also found to act as accumulators
of
elements, such as trace metals and sulfur (Ferrey et
al. 1973; Nieboer and Richardson
1981; Puckett
1985;
Tyler 1989).
The pollutant
Since there are no standards for conducting lichen
floristic studies or for calculating the completeness of
flora,
difficult to determine
the absence of
due to anthropogenic influences or to the
species
lack of
comprehensive baseline survey. We should
also consider the possibility that changes in lichen
communities are
function of changes in other
environmental
variables
that most directly
affects the health
is
(see Chapter 3).
the disappearance
Rydzak (1969) argued that
lichens from polluted
of lichen communities
sulfur dioxide (McCune
1988; Nimis et al. 1990). Lichens are often studied
a
waxy cutin, and consequently
of
cites at the turn of the century
was due to lowered relative humidity,
increased
temperatures, and more frequent wet-dry
the altered urban environments.
cycles in
This argument
is
a
and
if
such as soil, rocks, and the branches and boles of
vascular plants. Lichens lack an epidermis,
number of factors complicate the interpretation
of field and laboratory studies of lichen response to
air pollution. Recent studies of lichens in NPS units
have raised questions about the completeness of
species lists and the ability of field technicians to
perform follow-up surveys in subsequent years to
determine the existence of trends in species diversity.
a
ecological
and biological substrates
is it
is
extremely diverse floral group, occupying
niches on varied physical
and
a
Lichens
1992; Showman
COMPLICATING FACTORS
and the algae supply
through photosynthesis.
and Rosentreter
1989; Nash 1989; McCune 1988).
A
nutrition
a
support to the organism,
Thomas
Hendricks
a
Lichens are
group of non-vascular plants
composed of fungal and algal species growing in
symbiotic relationship. The fungi supply structural
countered by evidence
of
the return
of
within
species to cities as air quality has
pollutant-sensitive
(Coppins 1973; Rose and Hawksworth,
improved
chambers, and identification
the fumigation
of
and quantification
lichen responses to air pollution
exposures.
Some concerns in identifying pollutant-sensitive
1981).
species in field gradient studies include
of sites along
DISCUSSION TOPICS
a
homogeneity
pollution gradient, determination
of
pollutant exposures along the gradient, and assessing
lichen responses to these air pollution exposures.
During the workshop we discussed the following
topics, developed in more detail in Chapters 2-7 of
Lichen Responses to Air Pollutants
this volume:
1.
2.
3.
Lichen floristics (development of lichen
species lists and estimates of their
completeness).
Identification of lichen species that are
sensitive to air pollutants and the
pollutant concentrations that might affect
these sensitive species.
Methods used to characterize the status of
lichen communities and the ability of
lichens to accumulate air pollutants.
Evaluation of the overall usefulness of
lichens as biological monitors of air
quality.
Listed below are the more general topics discussed
4.
This discussion
of
determination
covered subjects relating to the
of
responses
lichens to air pollution,
definition of unique symptoms, the variability of
symptoms among individuals,
can be realistically
and what symptoms
measured under field conditions.
For example, depression of photosynthesis may be
initial sign of air pollutant stress identified in
the
exposure studies, but it may be impractical
to
measure this parameter in the field in remote areas.
responses to air pollution
chlorophyll degradation, changes in
and respiration, alterations in nitrogen
We discussed possible
stress including
photosynthesis
fixation,
of
membranae leakage, accumulation
elements, and possible
reflectance,
toxic
changes in spectral
lichen cover, morphology,
community
structure, and reproduction.
during the workshop.
Lichen Biomonitoring Plots
Lichen Floras
We discussed the usefulness
Topics included consideration of the protocols for
conducting lichen floristic studies and methods of
quantifying the completeness of any floristic study
considering
the range
of
surveyed, the percentage
of
in community
composition,
(diversity
of
and elemental content. Objectives
(NPS unit,
National Forest) covered, and the amount of survey
time spent in each habitat. Other topics included
methods to determine changes in lichen floras by
comparing present lichen species with past species
lists reconstructed from herbarium specimens.
and abundance),
design must be well thought-out
this type of monitoring
and experimental
before setting up
plot.
Elemental
During this session we considered the
development of protocols for controlled
of
Analysis
natural conditions
and field studies conducted along
Important
identifying pollutant-sensitive
fumigations
pollutant
include
interactions,
considerations
for
species in chamber
exposure regimes, mixed
environmental
of
that might be
lichen tissue.
interest are defined,
it is
of
might be expected due to the accumulation
lichens under
(fumigations)
gradients.
of
content
important to understand the natural variability that
pollutant-exposure
pollutant
used to analyze the chemical
Once the elements
condition,
morphological
In this group we discussed protocols
Identification of Pollutant-Sensitive Species
and
structure
community
habitats and substrates
the land area
establishing
evaluating long-term lichen monitoring plots to
differentiate natural and air-pollution caused changes
conditions
elements from natural sources (e.g. sulfate from dust
and sea spray). Important
elemental analysis
identification
of
of
considerations
elements to be evaluated,
of analysis,
and control (QA/QC) of analyses,
herbarium samples, separation of
preparation,
for
lichen samples include
methods
quality
sample
assurance
comparison
anthropogenic
with
elements from natural-source
stable isotopes),
elements (e.g. use of
with samples of host
and comparison
substrate (bark, soil, rock).
Transplants and Biomonitoring Gardens
of protocols for
or transplantation of lichen
species. In these kinds of experiments it is important
We discussed
the development
of
elemental content, and evaluation
plots over time.
QA/QC information is essential to providing credible
data for regulatory proceedings.
Geographic Information Systems (GIS)
GIS has the potential to be used in the
of
of
active manipulation
development
to consider
air pollution, land-use, and storm
patterns. All of these abiotic variables can affect the
distribution of lichen species. GIS can be used to
performing
establishing
lichens
reciprocal
species transplants,
near pollutant
monitors for
studies, physiological
exposure-response
analyses to
evaluate effects of transplant shock, and comparing
macro- and microhabitat
conditions
map plot-level
species composition
Considerable
be done before
or toxic
development
we can use spectral
to measure lichen responses or to provide
QA/QC on visual estimates of lichen cover and
condition.
Sampling Design and Statistical Analysis
Topics covered during
statistical
of
sampling
analysis
the discussion
These collections
elemental concentrations
collected
with herbarium
in pre-pollution eras.
If
samples
such comparisons
are to be made, it is important
to obtain adequate
Controlled
should be performed
to determine the
degree to which preserved specimens
can be used to
evaluate changes in pollution deposition over time
and to identify signature elements of anthropogenic
deposition.
structure, elemental
research and monitoring.
Meteorology and Climate
We discussed ways that researchers can evaluate
trends in meteorology
Quality Assurance and Quality Control of
Data Collection
the need to develop QA/QC
to be used during the collection and
structure and vigor,
particularly
of
in
patterns, which may
affect lichen species and community
vigor. The usefulness
structure and
these databases depends on
their accessibility at local and regional scales. When
designing lichen fumigation studies, climate data are
ambient environmental
conditions.
Air Pollution Monitoring
We discussed
of data on community
and climate,
temperature and precipitation
useful to approximate
protocols
tools for comparing
structure, and
amounts of tissue for analysis.
Some important things to consider
in experimental design and analysis are stratification
of habitats, random versus systematic sampling,
frequency of re-evaluation, sample sizes, types of
data collected, and methods of analysis (e.g.
parametric, non-parametric, regression). This activity
is an important component of all aspects of lichen
analysis
are valuable
current lichen floras, community
to address spatial and temporal
variability in lichen community
content, and vigor.
included
designs and methods of
Collections
Herbarium
experiments
development
GPS are becoming
accessible to field researchers.
and cover,
spectral signatures with physiology
morphological
systems
more accurate and cheaper and are now reasonably
structure changes over time, and
element uptake in lichens.
work should
We discussed the use of land navigation
during subsequent observations.
We need to develop a field method to spectrally
analyze lichen species and communities that will be
analysis
Land Navigation Systems
systems (GPS) to locate
sample points and to allow for accurate relocation
Spectral Analyses
correlate
structure, vigor,
data on community
and global positioning
species.
compare community
maps
at host and
Monitoring air pollution exposures
at transplant sites with instruments should be done if
air pollution is to be linked to effects on the
able to quantify
designs, regional
and elemental content.
transplant sites.
transplanted
sampling
precipitation,
Air
pollutants
monitoring
of interest in lichen research and
include sulfur oxides,
nitrogen
oxides,
REFERENCES
and toxic metals. To estimate the dose or
fluorides,
to air pollutants,
exposure of lichen communities
the
following methods can be employed: instrumental
monitoring, passive monitors, or extrapolation of data
from distant sites using techniques
such as kriging.
Coppins, B.J. 1973. In Ferry B.W., M.S. Baddeley,
and D.L. Hawksworth
Lichens.
The Athlone
Great Britain.
Identification of Research Needs
We discussed
emerging
areas
of
and D.L. Hawksworth
(eds). 1973. Air Pollution and Lichens. The
Athlone Press, University of London, Great Britain.
environmental
concentrations,
389 pp.
Ferry, B.W., M.S. Baddeley,
research, including the effects of global warming,
elevated carbon dioxide
(eds). Air Pollution and
Press, University of London,
389 pp.
increased
UV-b, and nitrogen fertilization. Researchers need to
which of these stresses might have an effect
Gough, L.P. and J.A. Erdman.
of
Influence
1971.
consider
coal-fired
powerplant on the element content of
on lichen species and communities.
Parmelia
chlorochroa.
Larcher, W.
Cooperation with Other Research/Monitoring
Programs
Lichen monitoring
component
of
monitoring
programs
should be investigated
planned or ongoing
environmental
such as the USFS/EPA
II,
EPA
and the
Clean
Air
the
Status and Trends
Coordination of lichen monitoring
Network.
among agencies
will
McCune,
14.
Forest
USFS Global
Change Program, the NPS Global Change Program,
the National Acid Precipitation Assessment Program
Health Monitoring Program,
efforts
of
result in the comparability
Physiological
1975.
Springer- Verlag, New York.
B. 1992.
Health Monitoring,
as a
databases.
Bryologist
USDA
80(3):492-501.
plant ecology.
252
pp.
Lichen Communities.
Forest
Field Methods Guide. Section
Forest Service, Southeast Forest
Park, NC
Station, Research Triangle
Experiment
a
27709. pp. 1-6.
McCune,
B. 1988. Lichen communities
SO2 gradients in Indianapolis.
along O3 and
91:223-228.
Bryol.
Nash, T.H. 1989. Metal tolerance in lichens.
119-131
Pages
in A.J. Shaw (ed.), Heavy Metal Tolerance
in Plants: Evolutionary
Aspects. CRC Press, Boca
Raton, Florida.
Nieboer, E., H.M. Ahmed,
THE PROCEEDINGS VOLUME
The workshop participants
in-depth discussion
Richardson.
and development
into chapters to
in the proceedings of the workshop.
six working groups developed a detailed
be included
of
the
outline
of
in relation
chose six areas for
the chapter, identified
Each
Drafts
of
the
chapters were sent out for peer review and the
reviewer
comments
were incorporated
This document summarizes
of
much of the literature
of air
pollutants, and describes possible responses of
lichens to air pollutants, especially sulfur dioxide.
The final chapter of this volume attempts to pull all
lichens
this information
as bioaccumulators
together by describing
seven
air pollution scenarios, and presenting
studies that can help guide a researcher or land
manager in the design of a lichen monitoring
different
program.
to distance from a nickel
Sudbury, Ontario.
Lichenologist
Nieboer, E. and D.H.S. Richardson.
in S.J. Eisenreich,
ed.,
smelter in
5:292-304.
Lichens
1981.
Atmospheric
as
Pages 339-388
Pollutants
in
Ann Arbor Science Publ., Ann
Natural Waters.
Arbor, Michigan.
into the final
document.
on the use
Puckett, and D.H.S.
monitors of atmospheric deposition.
a lead author, and
assigned writing tasks and milestones.
K.J.
Heavy metal content of lichens
1972.
Nimis,
PL., M.
Castello, and M. Perotti.
Lichens as biomonitors
1990.
sulphur dioxide pollution
in La Spezia (northern Italy).
Lichenologist
22:333-344.
Puckett, K.J.
Temporal
1985.
element levels.
pp.
Lichen Physiology
case
of
variation
in lichen
211-225 in D.H. Brown, ed.,
and Cell Biology.
Plenum Press,
New York.
Rhoades, F.M.
1988.
Re-examination
of
baseline
plots to determine effects of air quality on lichens
and bryophytes in Olympic National
Report to National
Division,
Park Service,
Denver, CO.
Air
Park.
Quality
Final
Rose, C.I. and D.L. Hawksworth.
recolonization
1981. Lichen
in London's cleaner air. Nature
289:289-292.
Lichens,
1991.
bryophytes, and air quality in Pacific
wilderness areas.
Unpublished
Rydzak, J. 1969. Lichens
conditions
as indicators
of
Mariae Curie-Sklodowska,
the habitat.
due to industrial
of Flavoparmelia
emissions. J. Air
Assoc. 39:317-320.
of
the
Annals Univ.
C, 23, 131-64.
Showman, R.E. and J.C. Hendricks.
element content
Northwest
report, 10 pp.
1989.
Trace
caperata (L.)Hale
Poll. Control
and
Air
Pollution.
A
in
Almqvist and Wiksells
epiphytes and environment
cryptogamic
Stockholm
Ryan, B.D. and F.M. Rhoades.
ecological
Skye, E. 1968. Lichens
region.
AB, Uppsala, Sweden.
Boktryckeri
Thomas, A. and R. Rosentreter.
1992.
study of
the
123 pp.
Antelope
of lichens in Birch Creek Valley of
Idaho. Proceedings of the 15th Biennial Pronghorn
Antelope Workshop, Rock Springs, Wyoming, June
utilization
8-11, 1992.
Tyler, G. 1989.
Uptake, retention, and toxicity
heavy metals in lichens.
Pollution
Air
of
and Soil
47:321-333.
Wetmore, C.M. 1983.
1 National
Water,
Parks.
Service Contract
Lichens of the air quality Class
Final Report, National Park
CX
0001-2-0034,
158 pp.
Floristics
Thomas
H. Nash III,
Clifford M. Wetmore,
William Anderson, Charis Bratt, William C. Denison,
Sharon Eversman, Barbara Murray, Larry St. Clair
Floristic studies provide essential baselines for judging future changes.
This chapter documents the essential resources, collection protocol,
voucher
specimen
requirements,
identification
and database compilations
requirements,
procedures,
report
needed for thorough floristic
studies.
RESOURCES NEEDED
INTRODUCTION
Floristics may be defined
of
as a compilation
Literature
species present in an area and the distribution
A
for those species.
information
For aid in planning field work, literature should
on any previous lichen studies as well as
complete and
accurate species list is an ideal basis for
understanding
of
the flora
the area being studied.
gives ecological information
necessary for determining
information
concerning
the
resources
of
appropriate
information
The value of
the unit.
floristic studies in lichen biomonitoring projects
recently been reviewed
by Wetmore
Lichen floristic studies require
specially
It
about the unit, provides
species for biomonitoring, and provides
trained lichenologists
has
of
because lichens
Collection and
identification of lichens involve recognizing habitats,
substrates, and specimens in the field, and using
appropriate
collection,
curation,
and identification
Quality assurance (QA) and quality
(QC) standards must be met, as legal
Time constraints
challenges to a study are possible.
procedures.
climate,
and air
literature on the vegetation,
geology,
quality studies in the area.
These may be published
papers or reports on file in the study area
headquarters.
area
All
papers reporting
lichens from the
including monographs
must be consulted,
and
revisions.
Access to all
(1988).
the expertise
possess many unique characteristics.
be
available
revisions,
of
the pertinent
identification
of
may be available
monographs,
floras is essential for
and regional
In some areas, keys
for some of the lichens. If regional
the collections.
floras are not available,
it may be necessary to obtain
papers from other areas to aid identification.
well-equipped
lichen library
Only
a
is likely to have
adequate reference material for many study areas.
control
are frequently
part
of
report; for example,
must be completed
point source.
Herbaria
a study and its subsequent
a study and its reviewed
before construction
of
report
a polluting
For these reasons, beginners and
amateurs in lichenology
are not satisfactory
personnel
A
good lichen herbarium
identification
identified
herbarium
identification.
adequate peer review
authorities.
a
preferably
of
proposals
by other lichenologists.
and reports,
lichens.
is necessary for the
After
a lichen
been keyed out, it should be compared
floristic study of a management unit unless they
working directly in association with experienced
An additional QA/QC requirement is
lichenologists.
for
are
of
species has
with reliably
material for confirmation of the
The herbarium
should contain specimens
the group and/or duplicates
reference collections
identified
of
by authorities
specimens
seen
in
by the
Personnel
The researcher should be familiar with the other
who know the lichens of the
lichenologists
management unit or the taxonomic
there.
These lichenologists
help with problem
of
groups found
may be called upon to
and review of some
identifications
the identifications.
able to provide
Some ecologists
may also be
ideas on collection design and
helpful
soil types. Knowledge of site
For example, areas that
restricted to specific
field work.
history is also important.
have been burned in the past few decades may have
some lichens species that occur specifically on
Similarly, insect outbreaks will
charred substrates.
open up canopies,
change the local microclimate,
by other lichen communities.
of
initial reconnaissance
The managers of the study area (forest, park or
land management
unit) should be able to provide
maps, air photos, climate data and other information
needed for field work and report writing.
The potential
for
aquatic lichens should also be considered.
Sites of about one hectare in size in major habitat
types should be replicated
Site-Specific Information
and
provide extensive wood substrates that are occupied
if
possible.
the study area
Ideally, an
would
of potential sites in each major
procedure could then
be used to select the replicate sites within each
identify
a number
A
habitat type.
randomization
habitat type.
Tools
COLLECTION PROTOCOL
the
It is essential that well-trained professionals make
field collections.
Because of their relatively
small size, many lichens
may not be noticed by
with initial
in lichenology will not be sensitive to the
range of microhabitats in which lichens occur.
Poorly trained individuals are not sensitive to the
amateur naturalists,
and even individuals
training
subtle morphological
different
differences
species or populations
important to be aware
well
of
as the more obvious
that distinguish
in decline.
relatively
It is
obscure species as
ones, since some of the
obscure ones may be more sensitive to air pollution
(Wirth 1985).
A geological hammer and coal chisel are essential
for collecting rock lichens that are normally collected
with a sliver of the substrate attached. A hunting
knife and/or wood chisel are essential for collecting
from trees.
Lichens growing on loose soils require
prompt impregnation of the soil with a dilute
(i.e., water soluble) glue after removal to
insure that the specimen does not disintegrate during
mucilage
All
transport.
should be stored in paper
will quickly mold in plastic bags or
specimens
bags as lichens
other closed containers.
If
it is not practical
moisten umbilicate
examine reproductive
Initially, sampling design strategies should be
discussed with the appropriate federal land manager
and resource people.
representative
Sampling
should include areas
in site selection.
to consider
USGS topographic
Consequently, it is
the effects of topography.
maps are helpful.
One should
sample on north- and south-facing slopes as well as
in riparian areas. Cliff faces, both in exposed and
shaded localities,
are frequently
rich in lichens.
Geologic maps should be consulted
may be specific
limestone,
from
where different
their removal
hand lens is essential to
structures (i.e., isidia,
with crustose lichens
species may appear superficially
similar.
of all major vegetation types (e.g.,
etc.). Microclimate variation is important in
determining lichen distributions, and should be
important
A
etc.), particularly
perithecia,
alpine, spruce-fir forests, ponderosa pine forests,
considered
lichens to facilitate
from the substrate.
SAMPLING DESIGN
to determine elevations
USGS maps, then altimeters should be carried.
Precise locations may be determined with a portable
GPS instrument. A bottle of water is helpful to
as rock lichens
to substrate (e.g., serpentine,
basalt, etc.). Similarly, soil lichens may be
SAMPLING PROCEDURES
At each site, ecological
and structure
of
notes on the composition
the vascular
plant vegetation
should be made. The full range of
community
substrates within the sampling
investigated
site should be
to insure as complete
possible. For example, different
on different
tree,
tree
species, at different
under shaded overhangs
locations,
a species
lichens
list as
may be found
heights on the
versus exposed
on mosses versus mineral
soils, on
rock types (size as well as geological
and on wood or bark in different
composition),
stages
of decomposition.
Unless a species is extremely rare, sufficient
fill
to
specimens.
Furthermore,
a 4" x 5" card (a
size to fit a typical lichen storage packet). Replicate
samples are recommended
herbarium.
Although some species
correctly
specimens with "fruiting" bodies
use. That
public
specimen
for
must be entered
accessible herbarium to insure that
publicly
available
future. Contracts
consequently,
have agreed to receive and maintain the voucher
(apothecia,
specimens.
variation
morphological
changes often occur prior to the actual
disappearance
of
voucher specimens
if
deposited in the herbarium
site; frequently,
U.S. National
Specimens
they should be air
should be packed carefully
the study area, as well as in the
Herbarium
other duplicates
Institution).
(Smithsonian
for
responsible
the study usually
to other herbaria.
When many small subareas are sampled in the
to prevent molding.
insure that the more fragile
distributes
sets
for transport
to
course of
fioristic study of
voucher specimen
specimens are not
large unit, only one
required for each species
recorded in the large unit. However,
crushed.
of
specimen
the
investigator
agency may wish to retain
or the contracting
a
collection,
within
of
one day's drive
that duplicate
a
of
be
voucher
The lichenologist
specimens
will
of
is
at the time
dried as soon as possible
If
the collector.
specimens be deposited in another herbarium
strongly recommended
a
are moist
several hours may be required to
sample the species present.
of
is
It
is
prepared, then that set
Label collecting bags clearly with, at minimum,
substrate type, date, and location.
Collection
numbers should be assigned, if practical, in the field.
In general, at least an hour should be spent at each
adequately
a
the
It
species.
minimum,
specimens which document
is
the
air pollutant-induced
At
a
morphological
collection of voucher
fioristic study must
consist of one voucher specimen for each species
recorded.
only one set of
expected that
etc.) should be sought. Particularly within
the more common species, pay attention to potential
perithecia,
as
into
for lichen fioristic
studies should stipulate which herbarium or herbaria
the indefinite
are typically
characters are important to
many lichens;
identify
use, a voucher
into and maintained by an established,
in more than one
may be deposited
asexual, reproductive
responsible
so that representative
specimens
of
to be
must be both protected and made available
it
is
material should be collected
for fioristic studies must make adequate budgetary
allowance for handling and preparation of voucher
is,
it
different
each species from each subarea.
VOUCHER SPECIMENS
of
species taken from a study site at a particular
time
in an established publicly accessible
herbarium. A voucher specimen of a lichen
and deposited
documents
the presence
site at the time
specimens,
of
study.
appropriately
an essential part
of
of that species at the study
A collection of voucher
prepared and identified,
the quality
assurance
of
is
any
fioristic study.
Note that a voucher
actual lichen;
Photographs
must consist of an
specimen
it may not be a photograph.
add to the value
of
substitutes for voucher specimens.
Verification of a lichen's identity requires
microscopic and chemical information not obtainable
voucher
specimen may be of
immense value. For example,
voucher
specimen
provides
during litigation,
evidence
of
a
a critical
species at a given place and time. But a good
voucher
specimen
lichen in
is expensive
to prepare. Contracts
specimen
on the species, the volume
will
contain substrate
- bark,
of
vary from one to ten cubic
centimeters. In many instances, the sample
will
rock, moss, soil, etc.
- in
addition to the lichen.
If
from a photograph.
well-prepared
Selection of appropriate samples begins in the
field and continues in the laboratory. Selection
involves choices based on the quantity of lichen
needed, its quality, and its rarity. Collections from
more than one site or time must not be pooled for
voucher specimens. Ideally,
voucher specimen
fioristic study consists of enough
documenting
material to include several to many lobes or branches
and contains representative sexual or asexual fruiting
structures. Depending
a report but they are
not acceptable
A
Selection
one
a
is an actual sample
the lichen
is
specimen
a
voucher
a
A
rare, collecting even for voucher
specimens should be severely curtailed.
collections
scientific
should be limited
purposes.
In all cases,
to those required for
Processing
Some selection,
sorting, and processing
made in the field, but specimens
the laboratory
processing
opened even when its back is attached to a sheet.
may be
arrive in
normally
in paper bags and in need
of
Many herbaria, including
depends upon the growth form of the
lichen and upon the amount and kind
of
substrate
included with the lichen.
Often a sample
will
include
Smithsonian,
routinely
further
Identification
before they can be placed in paper
packets for long term storage. The nature of the
processing
the
repacket specimens to their own standards.
more than one species
of lichen. Where it is possible to do so, the species
of choice should be freed of contaminants (e.g. ,
separated from mosses, most detritus, other lichens,
retention of part of the substrate is
In some instances, especially with
crustose lichens, it will not be possible to eliminate
etc., although
Some lichens,
in
be identified
of
require the use
of
some
mainly
the larger ones, can
However, many lichens
the field.
a microscope
and chemical
analyses to determine the species. The problem
identification is compounded by the lack of
comprehensive
Consequently,
a
manuals, regional
an investigator
of
or national.
proposing
to undertake
floristic study should be required to establish:
preferable).
all associated species.
In general, it is undesirable
to rewet lichens in the
However, bulky fruticose or foliose
species require moistening with water followed by
pressing in a plant press to permit their enclosure in
a packet and to prevent subsequent fragmentation of
laboratory.
the specimens.
Take care that moist lichens are
pressed and redried promptly
to avoid spoilage. To
avoid alteration of the chemistry of the lichen, use
distilled water rather than tap water, which can alter
in the use of appropriate
microscopic and chemical techniques;
1 ) expertise
2) access to equipment
techniques;
for performing these
3) familiarity with the world literature on
lichen taxonomy;
4) access to a major library containing the
relevant literature; and
5) access to a major lichen herbarium.
reactions.
Specimens
growing attached to soil require
treatment to consolidate
TECHNIQUES AND EQUIPMENT
the soil. Often this process is
begun in the field. The soil is infiltrated
with an
which dries and binds it together.
Without this treatment, soil and lichen disintegrate
over time. Any process that adds extraneous solutions
to the sample, such as moistening or soil
identifications
consolidation,
published
adhesive solution
realization
should be constrained
that subsequent study
may require microchemical
by the content
of
of
by the
the specimen
tests that may be affected
the adhesive solution.
Lichens growing attached to fragments of rock,
bark, or soil require special attention to prevent them
from abrading each other in the packet. In some
herbaria, the individual fragments are glued to a card,
which is enclosed in the packet. In other herbaria, a
card is covered on one side with a layer of surgical
cotton. Fragmented samples will adhere to the cotton
but can be removed for study.
Herbarium packets are folded from sheets of 100%
rag papers in a variety of ways. A standard size is a
4" x 6" (10 x 15 cm) packet. A label is attached to
the packet (see Hale 1979). Some herbaria glue the
individual packets to standard herbarium sheets. In
this case the packet must be folded
so that it can be
Reference Materials
Reference
materials essential to lichen
are of several types.
One category
literature, including books, monographs,
and journal articles.
Another is
the
is
collection of
specimens in a major lichen herbarium.
A
third is
world-wide network of lichen systematists.
A substantial collection of literature relating to
lichen identification is essential to lichen
identification given the absence of comprehensive
In rare instances, an
regional or local manuals.
the
investigator's personal library may be adequate, but
in most instances, lichen identification requires a
major research library.
a large technical
library
It is by no means certain that
will hold those specialized,
and often foreign, books and journals needed for
lichen identification.
Among the journals, the last 25
years
of
The Bryologist and Lichenologist are
Additional important journals include
Bibliotheca Lichenologica, Bulletin of the British
Museum, Cryptogamic Botany, Hertzogia, Journal of
Nordic
the Hattori Botanical Garden, Mycotaxon,
essential.
Journal of Botany, Nova Hedwigia,
Lichen taxonomy depends heavily on determination
Opera Botanica,
and Smithsonian
of
the
abundance
Contributions to Botany. Some of
important floristic treatments are listed in the
of
appendix
Published descriptions,
with or without
illustrations, are often inadequate for reliable
Under those circumstances, the
identifications.
must have recourse to comparison
investigator
of
metabolites.
this chapter.
his
Some
(TLC).
The methods
Culberson
serious taxonomic
collections
of
study
collaborate
of
globally
letter or phone call to a colleague
efficient way to resolve
of
substantial body
not-yet-published
results.
An investigator
with
"gray" literature
of
send voucher
original
Authority
specimens to an expert other than the
One instance, mentioned above,
investigator.
is when there is a colleague
determinations
in
a
specializing
difficult group.
in species
The second
instance is when there is an actual or anticipated
and it may be
in the genus.
legal challenge to the identity
of
a species.
Such an
instance is likely to arise in cases where the species
in question has been recognized as rare and/or
An added drawback
external authority
and access to appropriate
There are two instances in which it is advisable to
this resource.
specializing
techniques
these
Use of an External
or verification, it is desirable in
many instances to send specimens of species in a
difficult genus to a colleague who is monographing
the genus. Unfortunately,
many of the most difficult
genera have not been monographed
TLC
floristic
An
For determination
that no one is presently
a lichen
proposing
equipment.
study should be prepared to demonstrate that he or
she can make use
(HPLC)
study should be prepared to demonstrate familiarity
A
to undertake a lichen floristic
proposing
liquid chromatography
may be used to support or better define the
is the most
difficult identification.
and Johnson (1982), and
the principal investigator.
High-performance
within this informal network.
is exchanged
investigator
a
specific
Standard techniques described by
(1972), Culberson
of
discretion
few lichenologists, and they
well as nationally. Often a
as
to identify
(lichen chemistry) should
White and James (1975) are recommended. Other
published methods of TLC may be substituted at the
have daily access to a lichen herbarium.
There are relatively
lichen
chromatography
necessary to obtain a species
determination.
However,
a large number
of TLC
lichen metabolite products
requires that the investigator
normally
may be
but increasingly,
techniques,
is determined by thin-layer
specimen from a remote herbarium just as one may
loan.
secondary
these metabolites
microcrustal
be considered
a book on interlibrary
of
simple spot tests or
or her collections with herbarium material determined
and often annotated by experts. In a limited number
of cases, an investigator may borrow a critical
borrow
unusual and distinctive
determined by relatively
chemistry
of
because lichens produce an
lichen chemistry
to sending specimens to an
is that it might be months and
sometimes years before one receives a final
endangered, but may also occur when a species has
Providing a modicum of funding to
specialists for specimen verification would alleviate
been established as having a specific
determination.
known pollutant.
suggests the use
this problem.
sensitivity
to a
In any such case, discretion
of
an external authority
to
substantiate the identification given by the original
investigator.
Equipment
Expertise
in both microscopy
product determinations
and lichen secondary
New Species
is necessary to adequately
During lichen floristic surveys of previously
identify lichens. Much of the structure used in
This is especially
identifying lichens is microscopic.
true
of
crustose lichens,
unexplored
will
but reference to microscopic
species.
tissue types are required for many foliose
and
a
proposing
"taxonomic
lichens
as well.
An
investigator
may eventually
species"
of
as new
can be assigned to
a numbered
that genus, e.g., Lecanora
Ultimately, these preliminary
in one of two ways.
Either the species is finally recognized as belonging
to some described but obscure species, or it is
described and published as a new species. In some
lichen floristic study must demonstrate the routine
availability of a research quality light microscope
with oil immersion capability.
Access to a scanning
taxonomic
a
electron microscope
that some lichens
that are initially perceived
Usually such collections
genus and treated in reports as
features, such as algal cells, asci, ascospores, and
fruticose
areas, it is inevitable
be found
species 1.
identifications
(SEM) is highly desirable and
be essential.
10
are resolved
QUALITY ASSURANCE/QUALITY
CONTROL (QA/QC)
instances, publication of the new species may be
undertaken by the original investigator,
will
instances it
of
group
by a specialist in that
In sending material to an expert to
be described
lichens.
as new, it should
be described
type specimens
returned to
join
but in many
be understood
that
The following are
specimens collected
the
and/or an adequate paratype will be
the other voucher
The
specimens.
management agency and the herbarium
specimens
housing the
2.
Ensure quality storage for specimens
field to ultimate herbarium. Storage
should be dry, cool, pest-free.
with reprints of the original
3.
Labels
labels are made from
4.
archival-quality
labels, it is important
5.
to find papers, inks, and
Investigators
and herbaria will differ to some
labels and in the arrangement,
for a commonly
used format.
on
to be printed
The following elements should be
considered
in the design of labels:
State, county, governmental unit.
2.
Name of lichen - the scientific (Latin)
name of the genus and species followed
by the appropriate authorities according
to international convention.
3.
Locality:
a. Site specific name, e.g., Smith Canyon
4.
Substratum
This section contains
of
written reports. Subsequent publication in peer
reviewed scientific journals is strongly encouraged.
6.
format to
Using the following
outline will insure that the information essential for
establishing a lichen biomonitoring baseline is always
review and analysis.
included:
1.
2.
was
Name of collector(s).
a. Date of collection
b.
programs.
on format and preparation
should follow a consistent
Reports
facilitate
growing.
5.
suggestions
Format
plant vegetation
on which the specimen
a
to an established
lichen biomonitoring
Ecological site characterization:
Physical features: slope, aspect, elevation,
rock or soil type, etc.
c.
for
complete set of vouchers for all
institutional
Where possible, back up this
herbarium.
set with a duplicate set in a regional
Deposit
specific
Latitude and longitude/ or township,
range and section/ or UTM reference/
or GPS notation.
Biotic features: vascular
appropriate
REPORTS
a.
b.
of all
and references.
Preparation and submission of effective written
reports is central to the establishment of meaningful
1.
b.
from
herbarium.
but see Hale (1979)
Measurements should
be metric.
a
care in species
by use
When appropriate, submit samples
verification to external experts.
species
printers that satisfy these requirements.
degree both on the information
adequate
techniques
Voucher specimens must be accompanied by
Ideally,
Ensure
determination
paper and printed with ink that does
not fade or chip off. If computers are used to print
with respect to
concerns
floristic study:
Maintain adequate control associating
specimen with its field data.
description.
adequate labels.
a
1.
author of the new species should provide the
voucher
QA/QC
in
3.
Abstract
a
brief overview of the project.
Introduction - a summary of current
lichen air quality biomonitoring literature
with any specific information about
previous lichen surveys in the study area.
Methods references
Collector's number.
-
a
description with adequate
and laboratory methods
of field
used.
Name of determiner.
4.
11
Site description - a brief description of the
major habitat types within the study area
together with detailed information about
A
each specific collection locality.
Results - the species list will comprise the
The
major portion of this section.
following information should be included
for each species:
A. Genus, species, and authorities for each
on "missing" species
E.
Comments
F.
Comments
on the occurrence
abundance
of pollution
on significant substrate
H. Comments
considerations
Comparison of historical information or
data from comparable sites with current
I.
C. Growth form.
data
D. Substrate information.
J. Evaluation of ecological
E. Relative abundance information (i.e.,
rare, common, abundant, etc.).
K. Assessment
herbaria
specimens
concentrations
This list is neither comprehensive nor
documentation.
of
of pollutant
data
in lichens
Pollution sensitivity status with
G. Deposition
species
and distribution patterns
taxon.
appropriate
and relative
sensitive
on growth form frequency
G. Comments
B. Collection site(s).
F.
endangered
species
should also be included.
5.
on rare and/or
D. Comments
general map of the study area, showing
the location of each collection site,
(i.e., name(s)
other appropriate
absolute;
of
included depending
and collection number(s).
of
characteristics
H. Notation of species found in earlier
items may be
the specific
upon
site and/or
a study
lichen flora.
studies.
I.
General
comments
concerning
disjunct
Review of Reports
distribution patterns, new distributional
records, unusual substrates, modified
growth forms, unusual reproductive
patterns, incidence of thallus bleaching
or necrosis, or unusual nomenclatural
All
of
Expanded reports may include elemental
analyses and community data, such as
transect surveys. (See Chapter 7 for
details on elemental analyses and Chapter
4 for community analyses.)
Taxonomic
of sensitive
keys and color photographs
indicator species may also be included.
sexual
These individuals could be chosen
board proposed
should be given copies
determine whether appropriate
of
below.
the project
techniques
procedures were followed throughout
and
the project.
They should also comment on the general
presentation and organization
of
the report as well as
offer any new or unique observations.
DATABASES
At
the simplest
level, a floristic database conveys
More complex floristic
all data about the plants
that grow in specific sites. When a written flora, or
manual, is produced, other types of floristic data,
what plants grow where.
databases may encompass
morphological
B. Comments on the general health of the
lichen community
of
technical
contract and any approved modifications.
The
primary responsibility of the reviewers is to
Conclusions and recommendations - This
section of the report is diagnostic in
nature and should be based on a critical
interpretation of the data contained in the
results section.
This section essentially
defines the baseline condition. An
appropriate combination of the following
may be included:
A. Comments on lichen species diversity
abundance
the study area.
from the national advisory
Reviewers
C. Observations on the occurrence
reports should be evaluated by two
expertise and familiarity with the habitats and flora
considerations.
6.
technical
peer reviewers who have appropriate
identification
and chemical,
possible
etc., that make
are also important.
The term database refers not only to the above
and
types
and asexual
of
data but also to the
gathered and stored.
reproductive structures
form in which they are
floristic database
The simplest
could be a field notebook containing
12
plant names and
collection information that is associated with each
name. With the advent of readily available
database software,
computers and sophisticated
However,
limitless.
and numerous,
standardization
limitless
of
data.
Taxonomists
Authority
State
and
others interested in floristic and associated data are
County
such standards, including minimum
developing
Land management
for data collected for floristic,
and collections management purposes.
requirements,
monographic,
Standardization
makes it possible
Easily mappable
hardware and
Minimum standards ensure adequate data
and flexibility in database design permits
special-purpose data collection while retaining the
minimum standards agreed upon for all floristic work.
Minimum standards for floristic data should
collection,
of
plants and the authors
of
names, as well as information on the localities
which
the plants
are found.
Latitude
Longitude
UTM grid or GPS notation
those
in
taxon-related
General habitat
and
Specific habitat
Taxon-related data that must be
specimen-related.
included in floras are scientific names at the rank of
genus and below as well as the authors
names.
Required
information
specimen-related
that is traditionally
of
Substratum
these
Elevation (meters)
data include the
included
of accuracy of mapping points
Degree
These data can be
placed in two general categories:
locality
Specific locality
software.
include the names
unit
Physiographic unit
to transfer data
between databases made on different
appropriate)
Infraspecific name
possibilities
different computer systems require the
of data collection in order to facilitate
transfer and exchange
(if
Infraspecific rank
the
for storing and presenting floristic data
possibilities
are almost
Authority
on specimen
Collector(s)
labels (scientific name and authority, locality of
collection, collector, collection number, collection
Collection number
date). With these data, it is possible to produce the
simplest kind of flora — a list of what plants grow
Date
where.
Determiner (if not collector)
Possible
standards for data collection for lichen
of collection
Herbarium where deposited
floristic studies may include:
Recently,
Names:
manipulate,
have been used to store,
seen from the list of possible
Genus
names and for specimens,
both sets
Species
of
data:
As can be
data standards for
one item is common
names.
scientific
to
This common
element relates the two data sets, since the two data
Authority(ies)
Infraspecific rank
computers
and report pertinent data.
With the advent of
files share common fields.
(if
appropriate)
sophisticated
relational
relate numerous fields,
Infraspecific name
databases, it is possible
but also enables complex
Authority(ies)
When different
same organism
analyses and reports.
names have been used for the
in the area covered by the flora,
necessary to keep track
Specimens:
of
An accepted name, usually taken
of North
Lichens (Egan 1987), is used in floras,
from an accepted list such as the Checklist
American
Species
13
it is
this information by listing
the synonyms.
Genus
to
which simplifies data entry
with synonyms used in the flora area listed also.
Other information that pertains to genera or species
are often also available
It was suggested that federal agencies arrange
agreements with and retain experts who may be
called upon for completing
(family, growth form, etc.).
and other lichen-related
and needed
assignments
verifications,
tasks such as identifications,
surveys,
work.
NATIONAL ADVISORY BOARD
LITERATURE CITED
The Floristics Group at the 1991 workshop
recommends
established
that a National Advisory Board be
Culberson,
to advise federal and other agencies on
including floristic
Advisory Board should
studies.
The functions
of
the
Journal
To provide technical
on biomonitoring
2)
Egan, R.S.
1987.
lichen-forming,
a
method.
113-125.
72:
Substitution
1982.
for diethyl ether in
chromatographic
the
method for
Journal of Chromatography
A fifth
checklist
238:
States
of
the
and allied fungi of
lichenicolous
continental United
and Canada.
Bryologist
90: 77-173.
Hale, M.E., Jr.
4) To provide liaison among lichenologists.
1979.
How
to
Know
the Lichens.
Dubuque.
5) To recommend research priorities. It is
suggested that the Advisory Board be
composed of five lichenologists
representing different regions of the
United States and various areas of
expertise, plus one representative from
each appropriate federal agency.
Suggested terms are three to five years,
with staggered appointments and no
consecutive terms.
Wetmore, C.M.
quality.
Lichen floristics and air
III, T.H. and V. Wirth
1988.
Pp. 55-65.
In: Nash
(eds). Lichens, Bryophytes and Air Quality.
Berlin and Stuttgart.
Bibliotheca Lichenologica
30.
White, F.J. and P.W. James.
microchemical
57:
Okologie
Advisory Board
in coordinating efforts
1985.
work among lichen
herbaria.
14
of
British Lichen Society Bulletin
Zur Ausbreitung,
Herkunft
und
anthropogen geforderter Rinden-und
Holzflechten.
of the country that will culminate
of a national lichen flora. This
new guide to
1-41.
Wirth, V.
the National
A
1985.
techniques for the identification
lichen substances.
parts
would include cooperative
tert. -butyl ether
lichen products.
(PSD).
in the production
and new
483-487.
3) To provide QA/QC control and help ensure
legally defensible work, especially with
regard to the Clean Air Act (CAA) and
Prevention of Significant Deterioration
from various
Chromatography
standardized thin-layer
qualified personnel for specific projects.
liaisons,
conditions
of lichen products by
C.F. and A. Johnson.
of methyl
To provide information concerning
In promoting
of
Culberson,
and protocol advice
projects.
could also be instrumental
Improved
1972.
standardized thin-layer chromatographic
be:
1)
C.F.
data for the identification
all lichen biomonitoring projects,
Tuxenia
5: 523-535.
Chapter
2
Appendix
Some important floristic treatments for North America
General
References:
Fink, B.
1935.
States.
For the Northeast:
Brodo, I.M.
The Lichen Flora of the United
Ann Aibor.
(Largely
York:
York
out-of-date for most
genera, but still the only source of information
for a
number of genera.)
Hale, M.E., Jr.
Dubuque.
1979.
(Limited
How to Know
for
Gowan,
American
The Macrolichens.
Arctic Lichens I.
California.
(Obtainable
The Bryologist.)
1-254.
(Limited to
Bryologist
as a separate reprint from
For the Southeast:
Lichens of
California Natural History Guides.
3:
The lichens of Fundy
Park, New Brunswick, Canada.
91: 255-325.
For the West Coast:
1988.
S.P. and I.M. Brodo.
National
New York.
Hale, M.E., Jr. and M. Cole.
of the Ottawa Region.
Club Special Publication
(Covers all species; obtainable from the
National Museum of Natural Sciences, Ottawa.)
For the American Arctic:
1984.
Lichens
1-115.
illustrations.)
J.W.
1988.
Ottawa Field-Naturalists
the
lower 48 states, but contains many useful
Thomson,
and Floristic Analysis.
(Contains keys to all groups.)
Brodo, I.M.
the Lichens.
The Lichens
Vegetational
State
1-330.
to selected macrolichens
of Long Island, New
New
Museum Science Service Bulletin 410:
1965.
A
Harris, R.C.
54:
York.
the macrolichens.)
1990.
(Published
Some Florida Lichens.
New
by the author, but available
through the New York Botanical
Garden.
It
contains valuable keys for all groups.)
For the North Central Region:
Moore, B.J.
C.M. 1968. The Lichens of the Black Hills
of South Dakota and Wyoming.
Publication
Museum.
Michigan State University Biology Series
1:369-452. (Contains keys to all groups.)
Wetmore,
Bryologist
15
1968.
71:
The macrolichen
161-266.
flora of Florida.
Site Characterization
Ken Stolte, Sherry Pittam, Roger Rosentreter
The growth,
and biological environments
primary
physical
chemistry,
include
influences
of lichens are influenced by the
in which they are found. The
and survival
reproduction,
physical
are insolation,
temperature, moisture,
soils, and wind. The biological factors that most affect lichens
the moisture
and chemical characteristics
of
host substrates and
biological modification of the physical environment. To differentiate the
effects of air pollutants from other physical and biological environmental
factors, it is necessary to quantify, through measurement or estimation, the
environs in which the lichen species are found. This chapter provides
detailed methods for describing the environments in which lichen studies
are conducted.
INTRODUCTION
of
The purpose
and document
is generally
(Bailey, 1991). The microenvironment
considered to be the environment immediately
this chapter is to identify,
chemical,
the physical,
adjacent to the earth's surface, i.e., the zone below
quantify,
and biological
the height
of
4.5 feet, the height
of
standard weather
factors at sites where lichen studies are conducted.
instruments. Microenvironments for lichens
These factors can be chronic,
highly
influencing
physiological processes, or acute, affecting immediate
They have a substanal influence on the
survivability.
condition,
growth,
and survival
of
lichens.
trunk surfaces and crevices,
environments.
and chemical
some evidence
that lichens
factors most likely to
The macroenvironmental
affect lichens are:
to changes in their
exchange, they are responsive
and anthropogenic
materials.
Since
lichens lack stomata and have no control over gas
physical
There is also
respond to changes in
•
insolation
•
wind (speed, frequency)
•
temperature (maximum,
(intensity
and quality)
minimum,
their host substrates (Prussia and Killingbeck, 1991).
The goal of evaluating site conditions is to
•
moisture (rain, snow, fog, relative
factors so that changes in
•
atmospheric
quantify biotic and abiotic
lichen species and communities
can be reasonably
seasonality)
due to air pollution
lichen condition
and chemical variables
•
sites.
composition
of
factors controlling lichen growth and
lichen communities
include geology,
air, and water (Oosting, 1956; Ferry et al., 1973).
Biological factors include host species, canopy cover,
and herbivores
important
(Skye, 1968). These factors are
at both macroenvironmental
and
microenvironmental
refers to the climate
scales. The macroenvironment
and soil characteristics
region which influence
the
of
(nutrients,
humidity)
toxins,
acidity)
that should be measured or estimated at lichen study
The physical
chemistry
The microenvironmental
separated from other environmental
causes. We describe physical
are often
habitats such as rock crevices,
specialized
a
nature of the vegetation
16
factors that may affect
are:
host substrate
o
topography
o
chemistry
o
stability
o
moisture
o
type
- rock
-
tree bark
-
mosses
-
herbs
-
anthropogenic
materials
tree
soil
canopy opening
o
pedogenic
soil)
TEMPERATURE
o
edaphic (plant and soil interactions)
atmospheric (regional gases and gases
changes within a plant
whose concentration
canopy, e.g., carbon dioxide,
(percent)
of
(structure and chemistry
temperature ranges have a pronounced
Ambient
ozone)
changes with elevation
Of
the electromagnetic
photosynthesis,
to plants for
photosynthetically
of
insolation
latitude, elevation,
quality,
Lichen species distribution
(PAR). The
reaching the earth is affected by substances
the earth. Consequently,
of PAR
intensity
with
the algal component
varies with air
a good correlation
important
1968). There is usually
between habitat-type for lichen
(Levitt 1980a).
Site Measurement of Temperature
at higher
and northern latitudes.
easy to quantify the maximum and
temperatures at a site for any time interval,
It is relatively
minimum
and for some lichen transplant studies quantifying
Site Measurement of Insolation
Precise quantification
multiple
by
or the fungal component or both
species and their heat tolerance
effect on some lichen species,
and becomes increasingly
involves
controlled
1953, in Skye
equally (Lange
and can reduce the
is usually
by 85% or more. Ultraviolet light
may have a deleterious
elevations
ice
extremes may affect either
temperature. Temperature
seasons, and topography.
Tree canopies alter insolation
of
high temperatures (50o C) can lead to
of plant tissue and protein denaturation.
Extremely
desiccation
in the atmosphere and the angle of incidence
Temperatures
can result in the formation
crystals within cells and subsequent cell plasmolysis.
light are called
active radiation
communities.
vegetation
below freezing
between 460 and 640 nanometers.
These wavelengths
insolation
spectrum available
of
distribution
interest is the portion of
particular
by vegetation
slopes; decreases
on mountain
of approximately 1.5o C for every 1000 feet of
increase in elevation have a marked impact on the
Insolation refers to the radiant solar energy that
reaches earth.
plants.
effects are demonstrated
Temperature
INSOLATION
of all
on the distribution
influence
of
annual
PAR
at a site
of
transmitted
PAR
the results.
lichen transplants at new sites may be due to
in temperature ranges between sites,
the year (Isebrands et al.
not feasible, and in most cases
1992). This is usually
even a one-shot measurement
of
interpretation
samples with an integrated
radiometer throughout
in the
For example, death of
temperatures at the site may be important
differences
in air quality. Daily
rather than to differences
is
extremes
of
temperature and relative
difficult. The intensity and duration of the insolation
can be quantified into broad classes of exposure
obtained with relatively
based on factors that decrease maximum
thermometers.
hygrothermographs
insolation.
humidity
can be
low-maintenance
or maximum-minimum
For floral, elemental, or community
The primary physical and biological site factors
needed to measure or estimate insolation are:
studies involving surveys or plots, where many sites
are visited briefly, it is not possible to measure
Physical
temperature differences
temperature in any meaningful
way. Relative
among sites and microsites
•
latitude
can be estimated from the following site
•
aspect
characteristics:
•
slope
Physical
•
elevation
• micro relief
Biological
•
plant community
type
forest type (density of crowns)
o understory vascular and non-vascular
o
latitude and longitude
•
elevation
•
aspect
•
topographic
•
snow line (depth; duration
position
Biological
flora
•
•
•
canopy strata (layers and total height)
17
canopy cover (percent)
of
cover)
•
ATMOSPHERIC ELEMENTS
plant communities
o
forest type
o
understory
When the air, precipitation, or substrate is
with anthropogenic chemicals, lichens
will be contaminated somewhat proportionally with
the same chemicals, especially the foliose and
flora
vascular
contaminated
MOISTURE
fruticose forms. The degree
Moisture has
effect on the vigor of
a substantial
lichens and the diversity
of lichen
Lichens can photosynthesize
threshold,
(e.g., SO ), or more indirectly
(Farrar
the host substrate.
through accumulation
ecosystems as the lichens decompose. Toxic elements
known or suspected to negatively affect lichens
include sulfur dioxide, hydrogen fluoride, nitrogen
oxides, metals (e.g. Cu, Ni, As, Cd, Pb), ozone, and
radionuclides. (See Chapters 5 and 7 for details.)
1973,
in Ferry et al. 1973). Moisture assimilated by lichens
comes from the atmosphere in the form of rain,
relative humidity,
depends
(e.g., toxic metals). Metals accumulate in lichens and
then become a source of additional metal input to
and they
regime of their
and substrate environments
atmospheric
of contamination
of the lichens.
state
Toxic elements can affect lichens by direct toxicity
communities.
and respire only when
hydrated to a species-specific
tend to reflect the moisture
largely on the hydration
fog, dew, and snow melt, and from
Lichens can persist for long
periods when dried to moisture levels as low as only
1 - 15%
of
their dry weight.
Site Measurement
Lichens are adapted to different moisture zones
within habitats, and their physiological rates reflect
their moisture zones. Most lichens that have a high
tolerance for heat also have a high tolerance to
drought. They are usually, but not always,
characteristic of hot, arid regions (Levitt 1980b).
Some shade-tolerant lichens have similar adaptations
to high heat and drought. Lichens appear to be more
tolerant of dry air pollutants when they are not
Site measurement
of Atmospheric Elements
of
most atmospheric
elements is
difficult and costly, and in most cases is limited
specific
toxic elements
of
concern at intensive
sites. Toxic elements may be derived from
anthropogenic
sources (e.g., power plants, metal
or from natural sources, particularly soil
particles. In some cases toxic elements, sulfur oxides,
refineries),
nitrogen oxides, fluorides,
measured directly
hydrated.
[e.g.
sulfation
and ozone can be
with passive pollutant monitors
and nitration
or passive ozone monitors
Site Measurement of Moisture
plates (Noel et al. 1989);
and Hisham
(Grosjean
of active or passive pollutant
of the following site
chemistry will give some
1992)]. In the absence
monitors, measurement
Moisture can be measured directly at lichen sites
where detailed quantification
is desired, as in
transplant and fumigation studies. Moisture
availability can be estimated from the following site
characteristics
information
and
on nutrient and toxic element deposition
at a site:
variables:
Physical
Physical
•
•
elevation
•
topographic
position
parent material chemistry
•
soil chemistry
•
type and nearness
(horizon gradients)
of pollution source(s)
•
snow depth and duration
•
air and precipitation
•
aspect
•
snow melt chemistry
•
slope
•
soil texture (clay vs sand)
wind (speed and relative humidity)
•
chemistry
Biological
•
symptoms
in vascular and non-vascular
plants
Biological
•
•
vascular
•
plants
o
flora
o
general foliar morphology
non-vascular
o
to
study
chemistry
plants
(leaf size)
plants
mosses
18
of
vascular
and non-vascular
SOILS
•
host exposure
o
Soils are important components of ecosystems and
have both direct and indirect
influences
on lichen
The physical and chemical
characteristics of soils are the results of both
communities.
consistent
relationships
types (wet, semi-arid),
o
distance to nearest bush
o
percent vegetation
cover
exist between
groups (tundra, desert, forest), major
major vegetation
climatic
in rows, dense stand,
ing, etc.
the
parent material and associated biological
inorganic
communities;
(isolated,
location
etc.)
o distance from host to nearest tree, build
DATA FIELDS AND DATA SHEETS
and major soil
groups.
grow on soils, absorbing
Some lichens
moisture and nutrients,
directly.
To aid in analysis of site characteristics, we
suggest the following data-fields, which produce two
soil
toxins, and other chemicals
Lichens growing on vascular plants may
also be affected by soil chemistry
as chemicals
forms: one to be addressed by the agency or group
the lichen
supporting
are
filled out by
1) and the
study (Part
second
lichen expert, the principal
absorbed by plants and leached out through bark or
to be
leaves. Even characteristics such as soil texture and
color may affect lichens through wind-blown particle
abrasion, water capacities, reflection of PAR, or
absorption of heat. Soil characteristics help quantify
(PI) conducting the study (Part 2). Part 1
information on the general
physical, chemical, and biological factors that should
be considered prior to conducting the field work. Part
of
the ecology
a site and reflect
biota as well as anthropogenic
investigator
details the background
past climate and
2 details the information
the
Site Measurement of Soils
of
modified
chemistry
•
general particle
•
color
•
available
•
bareness (percent rock and plant cover)
The information
size
constraints
1 is important
and potential
present condition
of the site and
for
associated work
The information
problems.
from Part 2 is useful for analysis
and describes
the
the
distribution
patterns of lichen species. It records current
conditions for reference in future studies, particularly
Biological
duff depth
in Part
study sites and determining
choosing
water capacity
ground cover
et. al. 1992). It has been
and expanded in this document for lichen
Region (Jensen
field studies.
(pH, toxins, nutrients)
•
•
at
the sites during
Northern
Physical
that should be collected
field work. The format for
field data collection form (Part 2) comes from
ECODATA procedures developed by the USFS,
each
pollution.
the
•
the
long-term
analysis.
monitoring
for community
or elemental
Part 2 can help differentiate
the effects
of
air pollution on lichens from the effects of natural or
anthropogenic
CASE STUDY EXAMPLE OF
MEASUREMENT OF SITE
CHARACTERISTICS
Numerous
site variables
Part
This information
were measured or
Investigator
estimated by Skye (1968) to evaluate the effects of
air pollutants from Stockholm, Sweden on lichen
species and communities.
considered
very important
included
1968).
The
of
•
age
condition of
condition of
•
including
host plant
the tree
crown
the bole
o
growth
o
length of trunk
o
bark appearance
may be provided
by the contracting
to the Principal
agency, and should be
in requests for bids. It is intended to give a
necessary resources needed to conduct
following site data were recorded:
•
1
general overview of the area where the lichen studies
are to be conducted and to aid the PI in calculating
Host substrate was
(Skye
influence.
habit
19
budgets, personnel,
logistics,
the
study,
and safety.
Part 2
that the Principal Investigator
We recommend
lichen studies are conducted.
of
determine which
It is left up to
PI
the
study.
USDA
Forest Service's
for each site where
provide the following information
most important or relevant for a particular
These data fields were modified from the
ECODATA
method (Jensen
et al,
U.S. Forest Service, Northern Region, 1992).
to
the following 106 data fields are
PART I
MANAGEMENT UNIT
3.
c.
Managing agency
Purpose of study
d.
2.
b.
Heading
1.
Unit name
a.
Geographic
location of study
b.
B.
1.
State
2.
County
Approximate boundary of unit by Lat./Long.
and description
appropriate).
etc.
c.
Types of known pollution and sources
Location of nearest mechanical air quality
monitoring station and type of monitor(s)
Locations of known pollution sources relative
to site
(if
3.
Length of growing season
Range of elevations
Mean annual T°, humidity, insolation,
Air chemistry
5.
A.
D.
environmental factors
material
Geology/parent
Soils (short description or soils map)
Physical
Biological factors
Biome type(s)
2.
Significant disturbance
a.
Natural
1.
C.
Anthropogenic
Unique plant/animal
3.
b.
Landform(s)/topography
Terrestrial
history
a.
species
b.
Aquatic
General climatic factors
a.
Annual precipitation
E.
LICHEN SITE-SPECIFIC
-
Key
ID -
Record Identifier
AG - Agency
(2
Field
1
Fields
1-8 -
FA
FR
FS
FW
characters)
Enter one of the following codes to describe the
agency conducting the study:
CODE
DESCRIPTION
AR
Bl
BL
CH
FG
Agricultural Research Station
Bureau of Indian Affairs
USDI, Bureau of Land Management
NC
NP
PV
SC
SL
UR
OT
USDI, Fish and Wildlife Service
The Nature Conservancy
USDI, National Park Service
Other private lands
USDA, Soil Conservation Service
Department of State Lands
University Research
Comments,
Other (explain
Form GF)
Enter the two-character Postal Service abbreviation
for the state
which the plot
located (e.g.,
Montana = M I)- Use
when the state
unknown. For countries other than the U.S., enter
Comments.
two-digit country code and explain code
characters)
NF - National Forest
develop their own coding conventions for other land
units, e.g., National Parks,
this subfield - explain
Comments.
numeric)
in
Enter the number corresponding to the U.S. Forest
Service's National Forest being sampled (See USDA
Forest Service for National Forest codes). Users may
20
in
Field
(2
in
a
is
N
U_
is
in
-
(2
2
- St - State
USDA, Forest Service (Air Resource Managei
USDA, Forest Service Research Station
USDA, Forest Service (ECODATA Plot)
For additional codes coordinate with the USDA,
Forest Service (Northern Region, Regional Office,
Range, Air, Watershed and Ecology Staff Unit).
Champion International Lands
State Fish and Game Department
3
Field
DATA
in
PART
Any other unique features
II
4.
Field
4 -
Yr - Year
(2
Enter the last two digits of the calendar
enter 2 Q)
numeric)
year (e.g.,
for 1990,
5 -
Field
Pit - Plot Number
(3
Enter sequential plot numbers for each examiner
through the calendar year, up to 999. If one examiner
samples more than 999 plots in one year, a new
examiner code should be assigned and the individual
numeric)
encouraged
Example - Fields 1-5: A USFS plot (AG=FS) in the
Northern Region of Montana (St=MT), on the Helena
NF (NF=12), during 1986, by Hike Lichens, which is
the 45th plot for I. Lichens in calendar year 1986,
would be entered as:
1 Ag
5 Pit
6 - Mo - Month (2 numeric)
Enter one of the following codes to describe the
calendar month of sampling:
Field
ES
MONTH
2 St
Q45
04
05
April
May
06
June
July
07
CODE
to take a vacation.
MI
01
January
10
02
February
March
11
August
September
October
November
12
December
08
09
03
Field
7 - Day (2
Field
8 - Name
Enter the calendar
numeric)
- Name of Examiner (20
characters)
3NF12
4Yr SS
day of sampling.
Enter the first initial followed by a period (.), then the
last name of the person doing the majority of the
species identification, data measurements, and
evaluations
(e.g., I. LICHENS).
SAMPLE SYSTEM DATA
Fields 9-13 - Sample Forms - Codes for Sample
Forms Collected at a Plot (Up to five 2 -character
fields)
Various types of sampling procedures may be used
to describe characteristics
of the site being sampled
(e.g., community grid, lichen line intercept cover).
Identify the different types of sampling methods used
at the site by entering up to 5 of the following codes
(Note: The coding conventions used below are
suggested for some of the more common lichen
studies; develop additional codes as needed and
CODE
DESCRIPTION
LF
LC
LE
LT
LG
LP
LB
LX
Lichen
Lichen
Lichen
Lichen
Lichen
Lichen
Lichen
Lichen
floristics
community
elements
transplant
gradient
plots
biomonitoring garden
fumigation exposures
Example: A macroplot on which lichen community
lichen chemistry analysis were done, would be
entered as:
explain in Comments):
Sample
Forms: 9
LC.
10
LE
11
and
12
13
The above indicates that Lichen Community
and samples of lichens for Lichen Elements
(chemistry) were conducted at the site.
21
analysis
Field 14 - Unit - Unit of Measurement
(1
character)
Note:
Lichen methods, whenever possible, will use
metric (M) units for all standard inventory and
monitoring projects.
Enter one of the following codes to describe if the
units of measurement entered are English (E) or
Metric (M). Either type of measurement may be used
at a plot; however, metric is preferred.
Field
15 - Permanent Plot ID (15 characters)
Enter the Key ID recorded in Fields 1-7 if this plot is
to be used as a permanent plot. If the plot sampled
is a repeat measurement
of a permanent plot, enter
the Key ID corresponding
was sampled.
Required for permanent
to the fiisl time the plot
Field 16 - PRI - Plot Remeasurement Interval (2
numeric)
For a permanent plot, enter the number of years
measured last year; Q 2 means it was measured 2
years ago; Q 9 means it was measured 9 years ago,
and : 1 means the remeasurement interval since the
last measurement is unknown.
since the last measurement. For example, Q Q
indicates that the plot is a permanent plot and was
not yet remeasured; Q 1 means the plot was
17 -
Required for permanent
Plot ID (15 characters)
enter the Key ID for a plot to be used
as a comparison with this plot (e.g., lichen transplant
Field
plots only.
plots only.
Comparison
Optional.
If appropriate,
study).
Field 18 - Potential Vegetation Form - Potential
Vegetation Formation (2 characters)
Enter one of the following codes to describe the
potential vegetation
formation
DESCRIPTION
AQ
Aquatic
Non-vegetated terrestrial
(sand dunes, scree, rock)
CF
CW
HW
of your plot.
CODE
NV
BF
BW
SW
SA
SU
HA
HU
ML
OT
Coniferous upland forest
Conifer-dominated
wetland
X
Broadleaf upland forest
Broadleaf-dominated
wetland
Shrub-dominated
alpine
Predom. herbaeous (graminoid,
wetland
Shrub-dominated alpine
Shrub-dominated
upland
Herbaceous dominated alpine
Herbaceous dominated upland
Moss/Lichen-dominated
Other Type-specify in comments
Unable to assess
EXISTING VEGETATION CLASSIFICATION
Fields
- 22 - Existing Vegetation Strata
LF - Dominant Live Life Form
character)
Field
(1
19
19 -
CODE
DESCRIPTION
A
Aquatic species dominate
Broadleaf trees dominate
Conifers dominate
Herbaceous species dominate
B
C
Enter one of the following codes to describe the
dominant (based on canopy volume) live life-form
present on the plot. (Note: Dominant life form = life
form with the greatest canopy volume; i.e., height x
H
K
M
cover.)
P
S
O
X
22
DATA
(graminoid/forb/fern mixture)
Krumholz
Moss or lichens dominate
Agricultural cropland
Shrubs dominate
Other (explain in Comments)
Unable to assess
forb,
fern)
If
the dominant live life form
broadleaf (B) trees, use:
For other life form types, enter
(i.e., not
you are unable to make this
applicable).
assessment, enter an
this field.
ft
Low (< 2.5 ft average height) dominated
Medium (2.5 to < 6.5 ft average height)
dominated
Tall (6.5+
dominated
ft)
the dominant dead life form
broadleaf (B) trees, use:
If
characters)
Low shrub (<2.5
average height)
dominated
Medium shrub (2.5 to
6.5
average
height)
Tall shrub (6.5+
dominated
1.0 in. DBH) dominated
Seedling
Sapling (1.0 - 4.9
DBH) dominated
Pole (5.0 - 8.9
DBH) dominated
Medium tree (9.0 - 20.9 in. DBH) dominated
Large tree (21.0 - 32.9 in. DBH) dominated
Very large tree (33.0+ in. DBH) dominated
in
MT
For other types or when no dead life form exists,
enter
(i.e., not applicable).
you are unable to
this field.
make this assessment, enter an
<
X
CC - Live Canopy Cover Class
character)
Enter one of the following codes to describe canopy
cover (CC) of the dominant live vegetation life form
CODE
-
DESCRIPTION
and Lower Layer
must each have at least
percent canopy cover
within
the database.
layer to distinguish them
no single species has at least
percent cover within
percent
layer (yet collectively the layer has
cover), enter
representative code to describe the
dominant life form within that layer (e.g.,
E_
layer has
to indicate "not present."
no vegetation, enter
I
23
P-
ft
it
in
a
B
E
a
a
SHRUB, QBASS.EQRBlxIfa
5
A
Note: Fields 23 through 28 are used to describe
characteristics of the upper layer (above 6.5 ft),
middle layer (2.5-6.5 ft.), and lower layer (below 2.5
minimum of
percent canopy
ft.) vegetation zones.
cover must be present within
layer to distinguish
the database. Dominant and codominant species
5
5
If
in
5
Fields 23 - 28 - Upper, Middle,
Vegetation Zones
(<
X H M L N
Field 20:
No cover
9% CC)
Low cover (10-39% CC)
Moderate cover (40-69% CC)
High cover (70-100% CC)
Unable to assess
a
identified
in
(1
Field 22
If
ft)
TS
ft
ft
MS
in
LS
SE
SA
PT
ft
DESCRIPTION
DESCRIPTION
LT
VL
shrubs (S), use:
CODE
CODE
in
If
is
is
the dominant dead life form
conifer (C) or
(
is
If
the
canopy cover of dead stems and branches.
trees, the size class
dominant dead life form
chosen should have 1% canopy cover of dead stems
and branches.
is
- DSC - Dead Life Form Size Class
Enter one of the following codes to describe the size
class of the dominant dead life form on the plot (i.e.,
trees or shrubs).
the dominant dead life form
shrubs, the size class chosen should have 1%
If
< 1.0 in. DBH) dominated
Seedling
Sapling (1.0 - 4.9 in. DBH) dominated
Pole tree (5.0 - 8.9 in. DBH) dominated
Medium tree (9.0 - 20.9 in. DBH) dominated
Large tree (21.0 - 32.9 in. DBH) dominated
Very large tree (33.0+ in. DBH) dominated
X
LS
MS
21
SE
SA
PT
MT
DESCRIPTION
Field
DESCRIPTION
LT
VL
CODE
TS
(2
live life form is shrubs (S) use:
CODE
If
If the dominant
the size
in Field
the
the size
in
to describe
class of the dominant live life form specified
19. (Note: Size class is selected to describe
dominant layer component of a life form; i.e.,
class with maximum canopy volume.)
conifer (C) or
(
characters)
Enter one of the following codes
is
Field 20 - LSC - Live Life Form Size Class
(2
Field 23 - UL Dom Spl - Dominant Species in the
Upper Layer (8 characters)
Enter an eight-character code to describe the
dominant species in the upper layer (above 6.5 ft.
tall), if present. If no upper layer exists, enter N E
you are unable to make this assessment, enter X*
Field 24 - UL Dom Sp2 - Codominant Species in
the Upper Layer (8 characters)
Enter an eight-character code to describe the
codominant species in the upper layer (above 6.5 ft.
tall), if present. If no upper layer exists or exists with
no codominant species, enter N R If you are unable
enter JL
to make this assessment,
Field 25 - ML Dom Spl - Dominant Species in the
Middle Layer (8 characters)
Enter an eight-character code to describe
dominant species in the middle layer (2.5
tall), if present. If no middle layer exists,
you are unable to make this assessment,
the
- 6.5 ft.
enter M R
enter )L
If
Enter an eight-character code to describe the
codominant species in the middle layer (2.5 - 6.5 ft in
height), if present. If no middle layer exists or exists
with no codominant species, enter M P-» If you are
unable to make this assessment, enter 2L
Field 27 - LL Dom Spl - Dominant Species in the
Lower Layer (8 character)
Enter an eight-character code to describe the
dominant species in the lower layer (below 2.5
R,
no lower layer exists, enter
present.
tall),
you are unable to make this assessment, enter )L
Field 28 - LL Dom Sp2 - Codominant Species in
the Lower Layer
characters)
Enter an eight-character code to
codominant species
the lower
no lower layer
present.
tall),
no codominant species, enter
enter
to make this assessment,
If
N-
ft.
describe the
layer (below 2.5
exists or exists with
you are unable
E*
If
N-
If
if
(8
in
If
if
ft.
Field 26 - ML Dom Sp2 - Codominant Species in
the Middle Layer (8 characters)
)L
SITE DATA
(2
Field 29
- SpFea
characters)
-
Special Feature Information
CR
TA
a
Enter one of the following special feature codes to
describe the dominant environment of the plot. These
codes are hierarchical
design; consequently, you
C-,
may choose from the more generic codes (e.g.,
NS, E)or from the more detailed codes (e.g.,
N,
S
A A
in
SC
SB
NS
SW
WM
E).
DESCRIPTION
Not Applicable
Avalanche chute
With non-scoured surface
(soil not being eroded)
With scoured surface
(soil eroded, rocks exposed)
AS
CV
CL
CD
CI
(e.g., sedges, grasses)
Peatland (e.g., willows and birches)
Floating or quaking bog or fen mat
Swale (concave or bench area with no
surface water, but moist surface soil
moist
produces vegetation reflecting
SE
BE
Cave
Cropland
Dryland
Irrigated crops
environment)
Seep and/or spring
Concave bench with surface water creating
moist or wet soils that support vegetation
moist site
reflecting
Side Slope seep (not on bench or concavity)
Lakeside communities
a
AC
AN
PE
FL
SA
a
CODE
N
Sffl,B
Scree (sheet of stones/rocky debris mantling
slope)
Talus (rock fragments derived from and
positioned at the base of
cliff)
Streamside community
Stream bar community
Nonstream riparian communities
Shrub Wetland (e.g., willows and birches)
Wet meadow dominated by graminoids
SS
LA
24
TE
SN
Terrace (in valley bottom)
Snow catchment area (retains snow cover
2-4 weeks longer than 90% of the surrounding
areas of same aspect due to deeper snow
accumulation on a different physiographic
location, includes nivational hollows)
Cold air drainage or frost pocket
Wind blasted environments where vegetation
is maintained in a deformed state
Upslope warm air flow; area that is in the
direct path of afternoon warm upslope flow
from an adjacent deep valley bottom or a
thermal belt above a cold air inversion area
CA
WB
UW
Fields 30-32 - Landform
-
Geomorphic Landform
LANDFORM
1
Mountains
CODE 1
GM
Rl
RR
RS
RG
O
X
Windswept upper slopes that do not
accumulate snow and are relatively
dry due to snow loss compared
to adjacent areas
Ribbon forest
Ridgetop ribbon forest
Snow-caused
ribbon forest
Geologically caused ribbon forest
Other (Explain in Comments)
Unable to assess
Coordinate with the USDA Forest Service
additional codes.
LANDFORM 2
LANDFORM
CODE 2
Mountains
UM
MS
Cirques
CI
Trough Walls
TW
Trough Bottoms
TB
Avalanche
AD
Debris
Mountain Slopes
MS
Nivation Hollows
NH
Structurally Controlled
Complexes
SC
Benches
SB
Landslide
Deposits
LB
Undissected
BR
Structural
Breaks
SB
ST
Stream Breaklands
Dl
Walls
WA
FL
Undissected
UD
Dissected
Dl
Undissected
UD
Dissected
Dl
Undissected
UD
Dissected
Drainage
Hills
HI
RU
Rolling Uplands
Dl
Heads
High Relief
Low Relief
Valleys
Other
VA
Moraines
Kames, Kettles
MO
KK
Stream Bottoms
SB
Alluvial Basins
AB
Fans, Toeslopes
FT
Terraces
TE
O
Not Applicable
N
Unable to Assess
X
UD
Dissected
DS
Dip Slopes
Breaklands
CODE 3
PL
Plateaus
Structural
3
RT
Ridgetops
Mountain Slopes
Floors
Unglaciated
for
general-to-specific landform settings following Holdorf
and Donahue (1990). The first two-character field (i.e,
Landform 1) is always recorded; descriptions of
Landforms 2 and 3 are optional, yet highly
recommended.
(three 2 -character fields)
Enter one set of the following codes to describe the
landform where the site/plot is located. The codes
presented are hierarchical in that they describe
Glaciated
WU
25
DH
HR
LR
Fields 33-35 - PMat - Surficial Geology (three
fields)
*These types indicate situations where more than one
type of parent material may be influencing plant
growth. If appropriate, further specify the types of
parent material present on the plot by listing
appropriate rock type codes in the Rock Type 2 and
3 fields. For example, a site with volcanic ash over
sandstone would be entered as: M X/A £/§ A.; a site
with mixed sedimentary rocks such as limestone and
dolomite would be entered as: M S-/L 1/Q Q^
2 -character
Enter up to three of the following codes to describe
the type of parent material present at the plot. The
first code is required, the second code is highly
recommended, and the third code is usually specific
to individual project needs.
If an undefined third
code is used, keep a listing of coding conventions
and describe such codes in Comments.
ROCK TYPE 1
SEDIMENTARY
METAMORPHIC
GNEOUS
ALLUVIUM
ROCK TYPE 2
CODE 1
SE
ME
IG
AL
MIXED ALLUVIUM*
MA
MIXED SEDIMENTARY*
MS
MIXED METAMORPHIC*
MN
MIXED IGNEOUS*
Ml
MIXED FROM MORE THAN TWO ROCK TYPES*
MX
GLACIAL TILL*
GT
ASH*
LOESS*
AS
LI
Dolomite
Sandstone
DO
Siltstone
Shale
SI
SH
Conglomerate
Other type
CO
OT
Argillite
Siltite
Quartz ite
Slate
AR
SAND*
SA
O
NOT APPLICABLE
N
UNABLE TO ASSESS
X
SL
Schist
Gneiss
sc
Other type
OT
Basalt
Andesite
Diorite to gabbro
Latite
Quartz monzonite
BA
AN
Trachyte & syenite
Rhyolite
Granite
Welded tuff (Tufa)
Scoria (porcellanite)
Other type
TS
GN
Gravelly alluvium
Sandy alluvium
Silty alluvium
(Explain
26
SA
SI
QU
LO
OTHER
CODE 2
Limestone
in Comments)
Dl
LA
QM
RH
GR
WT
SO
OT
GA
AA
SI
Fields 36-37 - Pos - Plot/Site Position
(two 2-character fields)
Enter up to two of the following codes to describe the
position of the plot or site. The codes presented are
hierarchical in that they allow general (i.e., Position 1)
to specific (i.e., Position 2) descriptions of plot/site
position.
Moderate
Bottom (<100
Valley
ft.
PLOT POSITION
Narrow Valley
1
wide)
Bottom (100-300 feet wide)
PLOT POSITION
CODE 1
NV
MV
CODE 2
2
SC
Stream channel
Stream bar
Levee (narrow flood plain)
Colluvial deposit (fan)
Terrace
SB
LE
CF
TE
SC
FP
Stream channel
Flood plain
Abandoned meander
AM
OX
Oxbow
Backwater slough
Terrace
Alluvial fan (toeslope)
Other type
Slopes
Benches
MS
BE
SH
Ridges
Rl
Lower slope (fan skirt)
Mid-slope
Upper-slope
Short slope, neither upper nor lower
LS
Breaks
N
O
Not Applicable
Unable to Assess
X
Other
BK
NW
wide)
Wl
wide)
wide)
NW
Wl
Narrow (<100
Wide (>100
ft
wide)
NW
wide)
Wl
wide)
FL
Level to rolling plains
Upland ridge
Upland swell
Upland knoll
Upland breaks
River breaks
Plateau, mesa tops
Toe slopes, alluvial/colluvial
(Explain
27
MA
US
SS
Narrow (<100
Narrow (<100
RU
MS
US
Wide (>100
in
Badland
Uplands
BS
TE
AF
OT
LS
Wde (>100
Rolling
OX
Lower slope (fan skirt)
Mid-slope
Upper-slope
ft
Shoulders
AM
ft
Mountain
AF
SC
FP
Stream channel
Flood plain
Abandoned meander
Oxbow
Backwater slough
Terrace
Alluvial fan (toeslope)
Other type
ft
Fan
WV
ft
Alluvial
Bottom (>300 feet wide)
ft
Wide Valley
BS
TE
AF
OT
Comments)
UR
US
UK
UB
RB
fans
PL
TS
Field 38 - VerPS - Vertical Plot Shape
Enter one of the following
slope shape perpendicular
at the plot or site:
CODE
(1 character)
S
codes to describe the
to the contour of the slope
R
D
P
U
0
N
X
Field 39 - HorPS - Horizontal Plot Shape
(1 character)
CODE
S
Enter one of the following codes to describe the
slope shape parallel to the contour of the slope at
the plot location:
R
D
P
O
N
X
Field 40 - Elevation - Plot Elevation
Field
41
-
Slope Percent
(3
Straight or even
Rounded or convex
Depression or concave
Patterned (microrelief of hummock and
swales within several feet)
Undulating pattern of one or more low
relief ridges or knolls and draws within
plot area
Other (Explain in Comments)
Not Applicable
Unable to assess
DESCRIPTION
Straight or even
Rounded or convex
Depression or concave
Patterned (microrelief
swales within several
Undulating pattern of
relief ridges or knolls
azimuth of the plot's
Enter the declination-corrected
slope aspect to the nearest degree. Enter flat areas
as Q Q Q
Accuracy Standards - + 5 Percent.
Enter the average percent slope of the terrain on
Enter flat areas as
which the sample plot is located.
numeric)
Accuracy
(2
and
plot area
Other (Explain in Comments)
Not applicable
Unable to assess
QQQ.
Field 43 - EroStat - Erosion Status
of hummock
feet)
one or more low
and draws within
Enter the elevation of the plot above Mean Sea Level
in meters or feet. Use a topographic map or altimeter
to estimate elevation.
Accuracy Standards - + 100 meters
(5 numeric)
- Aspect - Slope Aspect (3 numeric)
Field 42 - Slope
DESCRIPTION
CODE
characters)
ST
Enter the most appropriate code based on indicators
of soil cover, sheet erosion, rill erosion, bank stability,
gullying, etc. Instability includes situations where
human activity has increased the rates of
disturbances above that which would occur naturally.
X
UC
UD
28
Standards
- + 5 Percent.
DESCRIPTION
Soil surface stable and no evidence of
accelerated erosion.
Unable to assess because examiner cannot
determine stability or instability compared to
undisturbed conditions.
Soil surface is unstable because of compactic
(weight per unit volume natural).
Soil surface is unstable because of
displacement and/or churning of the soil.
Field 44 - EroType
-
Erosion Type
(2
GE
characters)
DE
WE
SC
SL
Enter one of the following codes to describe the
dominant erosion process present on the plot.
CODE
N
SE
RE
DESCRIPTION
TD
SD
Not Applicable
Sheet erosion
Rill erosion
SP
Fields 45-52 - GCov
2-number fields)
O
X
CLASS
CODE
Splash erosion/soil crust
Other (Explain in Comments)
Unable to assess
CLASS
RANGE OF CLASS
MIDPOINT
Bare Soil ( 1/16 in. diameter soil
(2
0 cover
> 0 - < 1% cover
03
1 - < 5% cover
10
5 - < 15% cover
20
15 - < 25% cover
25 - < 35% cover
35 - < 45% cover
45 - < 55% cover
55 - < 65% cover
65 - < 75% cover
75 - < 85% cover
85 - < 95% cover
95 - 100% cover
30
40
50
60
70
80
90
98
Accuracy
Standards
0
0.3%
3.0%
10.0%
20.0%
30.0%
40.0%
50.0%
60.0%
70.0%
80.0%
90.0%
97.5%
- + 1 Class
Note: Percentages for the eight ground cover
categories should total approximately 100 percent.
Since cover classes are entered, the sum of the
class codes may vary from 90 to 110 percent.
and 20 percent, e.g., 30 percent is a very high value
that is rarely encountered in the Northwest U.S.
FLC - Fuel Loading Class
01
00
taken up by the live basal or root crown portion of
vascular plants; this includes live trees, Lycopodium,
and Selaginella.
This is not foliar cover of plants;
typical values for basal plant cover range between 10
-
development
Use the following standard cover class codes to
quantify ground cover by the preceding categories:
particles).
Field 46 • Gr Gravel (1/16 - 3 in. diameter).
Field 47 - Ro • Rock ( 3 in. diameter).
Field 48 -• LD ■Litter, Duff and Ash: charred and
uncharred litter, duff and scat; litter includes freshly
fallen leaves, needles, twigs, fecal material, bark, and
fruits; duff is the fermentation and humus sections of
the organic layer.
Field 49 - Wo - Wood: charred and uncharred dead
wood that is greater than 1/4 in. diameter. This
includes standing dead trees.
Field 50 - ML - Moss/lichen/fungi/algae
Field 51 - BV - Basal Vegetation: the soil surface
Field 53
Slump
Terrace
Slide
Field 52 - Wa - Water: that portion of the plot's area
which is covered by standing water at the time of the
sampling.
Ground Cover (eight
Enter a cover class code from the list below for each
of the following ground cover categories to indicate
cover at the soil surface plane.
(Note: Foliar canopy cover above the soil surface
plane is not considered to be ground cover.)
Field 45 - BS
Gully erosion
Deposition
Wind erosion
Soil creep
numeric)
Fuel loading classes are from the fire behavior fuel
models of Anderson (1982) and Albini (1976)(both
cited in Jensen et. al. 1992). Enter the appropriate
code from the following list. Classes are general and
describe the fuels that will drive the fire behavior
model. If you are not familiar with this system be
sure to reference Anderson (1982) and obtain
assistance from a qualified fire behavior specialist. If
you do not have Anderson's (1982) publication or
have not been trained to identify fuel loading classes,
CODE
DESCRIPTION
00
Unable to assess
Fine, porous and continuous herbaceous
fuels of grasslands, savannas, grass-tundra
01
02
03
04
enter Q Q in this field.
05
29
and grass-shrub types.
Fine herbaceous fuels with some litter and
dead stemwood in habitat types with open
shrub and forest overstories.
Tall, thick graminoid-dominated
stands.
Forest or shrub stands with a continuous
overstory that contain much flammable
woody material.
Forest or shrub stands with light surface
fuels and slightly flammable shrub and woody
fuels.
06
07
08
Open forest with shrubs or shrubs that have
moderate amounts of flammable woody material.
Closed forest stands and understory shrub
layer with flammable materials in both layers.
Closed conifer stands with low flammability
09
12
Closed stands of pine with a thick litter layer.
Closed forest types with heavy fuel loading
of down woody material.
Light logging slash, varying in continuity.
Moderate, continuous logging slash.
and a compact
13
Heavy, continuous
10
11
litter layer.
Field 54 - FD - Average Fuel Depth
(3
value is averaged over the plot (e.g., envision a
sheet draped over the ground fuels and live and
dead shrub/herb layer on the plot, and determine the
average height of that sheet.) This calculation does
numeric)
Determine the average depth of all live and dead
ground fuels on the plot to the nearest 1/10-foot. Fuel
depth is the vertical distance from the bottom of the
litter layer to the highest fuel particle (below 6.5 ft.
height) that is capable of carrying a ground fire under
moderate fuel moisture and wind conditions. This
Field 55 - Down
Diameter
(3
logging slash.
not include the crowns of overstory trees or any
species greater than 6.5 ft. height.
Accuracy
Log Dia - Down Log Average
Standards
- + 0.5-Foot.
but lying at an angle with the ground of no greater
than 20 degrees. Count all such dead woody material
that occurs below a height of 6.5 feet. Entries are to
the nearest inch.
numeric)
Enter the average diameter of the down woody
material. Down woody material is identified as any
sound or rotten log which is distinguishable as a log,
including logs not in direct contact with the ground
Accuracy
Field 56 - Dom. Layer Ht. - Average Height of the
Dominant Layer (3 numeric)
Standards
- + 6 Inches.
Enter the average height to the nearest foot of the
dominant vegetation layer on the plot (i.e., the one
with the greatest canopy volume).
Accuracy
Standards
- + 5 Feet.
VEGETATION DATA
Live Trees
Field 57 - DBH - Average DBH of Live Trees
numeric)
Estimate (from several measurements and an ocular
evaluation) the average diameter at breast height of
the dominant live tree layer (i.e., the tree layer that
has the greatest canopy volume).
(3
Accuracy
Field 58 - Height - Average Height of the
Dominant Live Tree Layer (3 numeric)
Standards
- + 6 Inches.
Estimate the average height to the nearest foot of the
dominant live tree layer (i.e. with the greatest canopy
volume).
Accuracy
Dead Trees
- + 5 feet
From several measurements and an ocular
evaluation, estimate the average diameter at breast
height of the upper dead tree size class layer (i.e.,
the size class in the tallest layer that has 1% dead
Field 59 - DBH - Average DBH of Dead Tree
(3
Standards
numeric)
stem and foliage cover).
Accuracy
Field 60 - Height of the Dominant Dead Tree
Layer (3 numeric)
Standards
- ± 6 Inches.
Estimate the average height to the nearest 5 feet of
the dominant dead tree layer (i.e., tallest layer that
has 1% dead stem and foliage cover).
Accuracy
30
Standards
- + 5 feet
Fields 61-67 - Tree Cover (seven 2-number fields)
75 - < 85% cover
85 - < 95% cover
95 - 100% cover
80
90
Estimate the percent canopy cover for trees as a life
form and by size class. This estimate is the
horizontal percent cover of the vertical projection of
trees and does not account for overlap. Do not
98
The above cover class codes
the following variables:
include trees less than 6 inches tall in this estimate.
The following percent cover class codes are used to
record tree cover:
0 - < 1% cover
03
1 - < 5% cover
10
5 - < 15% cover
15 - < 25% cover
20
30
40
50
60
70
Fields 68-71
25 - < 35%
35 - < 45%
45 - < 55%
55 - < 65%
65 - < 75%
-
overlap)
CLASS
MIDPOINT
Field
0
cover
cover
cover
cover
cover
Field 64 - Pole - Pole size cover (5.0 - 8.9 in. DBH)
Field 65 - Med - Medium size cover (9.0 - 20.9 in.
0.3%
3.0%
10.0%
20.0%
30.0%
40.0%
50.0%
60.0%
70.0%
DBH)
Field 66 - Large
Field
67 -
-
DBH)
VLarge
Large size cover (21.0 - 32.9 in.
- Very
large size cover (33.0+ in.
DBH)
Accuracy
Shrub Cover (four 2-number fields)
Estimate the percent canopy cover for shrubs as a
life form and by size class. This estimate is the
horizontal percent cover of the vertical projection of
shrubs and does not account for overlap. Enter the
appropriate percent class using the standard cover
class codes presented in Fields 61-67.
Fields 72-75 - Herb Cover
(four 2-number fields)
- 4.9 in.
DBH)
Standards
Field
Field
Field
69 -
Field
71 - Tal
68 -
70 - Med - Medium
Accuracy
Class.
Tot - Total shrub cover
Low - Low size shrub cover
6.5
Herbaceous Cover
+ 1 Cover
tall)
- Tall size
Standards
(< 2.5
tall)
size shrub cover (2.5 to
ft.
01
62 - Seed - Seedling size cover
- < 0.1 in. DBH)
( > 6 in.
63 - Sap - Sapling size cover 90.1
- +
shrub cover
Cover
(>
0 cover
Field
1
00
Tot - Total tree cover (do not include
61 -
6.5 ft. tall)
Class.
Field 72 Gram - Graminoid cover
Field 73 Forb - Forb cover
Field 74 - Fern - Fern and fern allies cover (includes
Enter the percent canopy cover for each of the
following herbaceous components using the cover
class codes presented in Field 61-67. (These
estimates do not count species overlap within an
herbaceous component.)
spp. and Selaginella spp.
Lycopodium
at < 6.5 ft.)
Field 75 - Moss - Mosses, lichens, fungi and other
bryophytes
Accuracy
Standards
- +
1
RANGE OF CLASS
are used to categorize
Field
ft.
CLASS
CODE
80.0%
90.0%
97.5%
Cover
Class.
DESCRIPTION
No disturbance
31
stable condition)
(if
mechanical or animal disturbance, 5-20
Low
percent of ground cover removed exposing bare
fire, most fine fuels burned,
soil and pavement;
some charring of 3+ in. fuels, woody plants
scorched but not burned to the ground; uneven
patch burning of duff and litter)
if
Enter one of the following codes to describe the
intensity of disturbance caused by mechanical
equipment (M), fire (F), and animals (A) on the plot:
(ground cover
in
CODE
L
Fields 76-78 - Ground Cover Disturbance: M, F, A
- Ground Cover Disturbance: Mechanical, Fire,
Animal (three 1-character fields)
N
DISTURBANCE DATA
QESQ
M
mechanical or animal disturbance,
Moderate
20-40 percent of ground cover removed exposing
fire, nearly all fine
bare soil and pavement;
fuels consumed, some consumption of 3+ in.
fuels, some woody plants consumed; patch
mechanical or animal disturbance, 40-100
of ground cover removed exposing bare
fire, many areas of exposed
pavement;
soil, many woody plants consumed, most
in. fuels charred and many consumed; fairly even
distribution of duff and litter consumption)
High
percent
soil and
mineral
if
distribution
of duff and litter consumption)
5,
6
4,
3,
2,
-
DO
DS
codes,
in
in
use evidence
DE
Human
Mountain Goat
Mountain Lion
Moose
Pocket gopher
ML
decreasing
MO
PG
PM
PW
RA
RT
TS
UB
WO
Wolf
Cattle
X O
Other (Explain
Comments
Unable to assess
Coyote
Deer
Consult the U.S. Forest Service
DESCRIPTION
No evidence
found
Antelope
Black Bear
Bear (species not known)
Bighorn sheep
Beaver
Porcupine
Pileated woodpecker
Rabbit
Raptor
Tree Squirrel
Upland birds
in
NO
AN
BB
BE
BS
BV
CA
CO
sheep
Elk
Grizzly bear
Ground squirrel
Horse
HU
MG
order of evidence.
CODE
Dog
Domestic
EL
GB
GS
HO
Enter the following codes to describe evidence of
animal use on the plot,
decreasing order of use
and effect on vegetation and site characteristics.
Evidence may include tracks, browse, caches, beds,
wallows, scat, antler rubs, nests, dens, etc.
Record
up to six animal
;
Unable to assess.
2
1,
Fields 79-84 - Animal Evidence
Animal Use Evidence (six -character fields)
if
(if
(if
CODE
Data)
for additional
codes.
HOST CHARACTERISTICS
General Characteristics of Host
Field 86
is
Estimate the average condition of the crowns of the
host tree(s), using an estimate of the live crown ratio
(0-100%) and crown density (0-100%) of the tree
crowns,
10% classes (listed below). For example,
tree with
100% live crown ratio has
live crown
extending from the top of the tree to the ground.
tree with
100% crown density has
Similarly,
completely dense crown with no sunlight visible
a
even
dead tree with no foliage has some amount of
crown density, even though live crown ratio would be
zero.
tree with no sunlight visible through the
crown density of 100%.
crown has
a
a
Use the following percentage classes
crown ratio and crown density:
through the crown.
Field 85 - LCR - Live Crown Ratio
CLASS
CODE
Estimate the average live crown ratio of the host
trees where the lichens were collected.
The live
crown ratio
the percentage of the tree bole that
supports living branches.
RANGE OF CLASS
is
00
32
10
>
20
0%
- 10%
- 20%
0
a
a
A
in
a
a
is
Estimate the average crown density of the host trees
The crown density
where the lichens were collected.
estimated relative to the amount of sunlight that
blocked out by the entire tree crown including the
bole, branches, foliage, and reproductive structures,
as outlined by the branches on the tree. Therefore,
Field 85-86 - Condition of the Tree Crowns - (two
2-number fields)
a
Crown Density
- CD -
1
1
Fields 85-106:
Plant
to quantify live
CLASS
CODE
RANGE OF CLASS
CLASS
CODE
RANGE OF CLASS
30
21
- 30%
70
61
40
31 - 40%
80
71 - 80%
50
41
- 50%
90
81 - 90%
60
51 - 60%
99
91 - 100%
Accuracy
Field 87-89 - Condition of the Tree Boles
Field 87
Field
GH - Growth Habit - (two 2 -character
Growth habit refers to the general morphology of the
tree trunk (bole).
Two fields should be recorded
describing the average condition of the tree(s) where
the lichens are collected or identified - one field
describes the average branching of the stem below
breast height (1.47 meters); the second field
describes the average inclination of the tree(s).
SV
DT
Nl
SI
Ml
HI
Standards
88 -
- + 1
Class
BTL - Bare Trunk Length
Estimate the length of the tree trunk in feet/meters
that is bare (from the ground to the beginning of
living branches) and open to colonization of lichens.
fields)
CODE
- 70%
Accuracy
Standards
- + 1.5 feet/0.5 meters
Field 89 - BAL - Bole Azimuth of Lichens
Record the common azimuth of the lichens found on
Record up
the tree boles, in classes of 60 degrees.
to six classes for lichens that are colonizing the
entire bole, i.e., lichens are found on all sides of the
tree bole.
CLASS
Branching:
Simple, undivided trunk
Divided trunk (2 or more stems)
CODE
00
06
Inclination:
No incline, + vertical trunk
Slightly inclined trunk (11 - 30 degrees)
Moderately inclined trunk (31 - 60 degrees)
Highly inclined trunk (61 - 90 degrees)
12
18
24
30
36
Accuracy
Fields 90-93 - BHL - Bole Height of Lichens
CODE
NA
Record the height of bole that contains lichens (all
species = Field 90), whether the lichens were
collected or identified from this entire area or not.
This refers to the maximum height of bole
00
05
10
colonization on the tree, even if it is restricted to one
azimuth. Estimate the height from the ground to the
highest point in the tree where lichens can be
observed from the ground.
Record height for all
species in Field 90, or record height for up to 3
individual species in Fields 91-93 (record species ID
15
20
25
30
35
40
45
50
in Comments).
10
99
Accuracy
33
RANGE OF CLASS
No lichens on bole
> 0 - 60 degrees
61 - 120 degrees
- 180 degrees
- 240 degrees
240 - 300 degrees
301 - 360 degrees
121
181
Standards
- + 1
Class
RANGE OF CLASS
Not Applicable
No lichens on bole
0-0.5 m above ground
0.6-1.0 m above ground
1.1-1.5 m above ground
1.6-2.0 m above ground
2.1-2.5 m above ground
2.6-3.0 m above ground
3.1-3 5 m above ground
3.6-4.0 m above ground
4.1-4.5 m above ground
4.6-5.0 m above ground
5.0-10.0 m above ground
> 10.0 m above ground
Standards
- ± 1 Class
Field 94-97
Lichens
BCHL - Bole Collection Height of
RANGE OF CLASS
CODE
NA
Not Applicable
No lichens on bole
00
05
Record the height on the bole from the ground where
all the lichen species were collected or identified (all
species = Field 94), or the height range on the bole
where up to 3 specific lichen species were collected
(Fields 95-97). These may be the same values as in
Fields 90-93 if all the lichen species are restricted to
the lower bole where collection or species
identification usually occur, from ground level to about
2.5 meters above ground.
0-0.5 m above ground
0.6-1.0 m above ground
10
1.1-1.5 m above ground
1.6-2.0 m above ground
2.1-2.5 m above ground
> 2.5 m above ground
15
20
25
30
Accuracy
Standards
-
+ 1
Class
Field 98-103 - Appearance of Bark
species is involved in collection or evaluation of
lichen species, record the bark characteristics of the
Record the general characteristics of the tree bark of
the host species, with particular emphasis on bark
morphology and bark color. If more than 1 host
3 most common
species.
Field 98-100 - BM - Bark Morphology
CODE
DESCRIPTION
Record the general morphological characteristics of
the bark of the 3 most common host tree species.
NA
BP
SP
NP
TF
SF
Not Applicable
Broad, flat plates
Small, flat plates
Of primary importance
is the general nature of the
bark (e.g., broad flat plates, deep fissures, etc.), and
the relative flakiness of the bark (e.g., flaky,
persistent, etc.). Record up to 3 codes for each tree
species, e.g., broad flat plates (BP), shallow fissures
(SF), with flaky bark (FB).
No plates, bark essentially
smooth
FB
LB
PB
Tiny fissures in bark ( 1.0 cm)
Shallow fissures in bark (1.0 - 2.5 cm)
Moderate fissures in bark (2.6 - 4.0 cm)
Deep fissures in bark ( 4.0 cm)
Flaky bark; freely exfoliating when touched
Loose bark; pulled off with slight effort
Persistent bark; firmly attached
CODE
DESCRIPTION
Record
NA
3 most common
WH
BN
Not Applicable
White
Black
Brown
RD
GN
TN
Red
Green
Tan
TS
SB
Tree stands (natural)
Surrounded by buildings
MF
DF
Fields 101-103 - BC - Bark Color
the general color (hue) of the tree bark of the
host tree species.
Combine up to
two codes to suggest lighter shades of any one hue,
with the first code indicating the predominant shade
of the hue.
Fields 104-106:
BL
Exposure of Host Species
Field 104 - HTL - Host Tree Location
Record the general location of the host tree(s) where
the lichens are collected, e.g., isolated trees, in
rows/orchards, dense natural tree stands, surrounded
Add additional
by buildings, etc.
and explain in Comments.
CODE
DESCRIPTION
N
Not Applicable
Isolated trees
Rows of trees
IT
RO
Field 105 - DNLO
Object
codes if necessary
Distance to Nearest Large
Estimate the average distance to the nearest object
that is greater than 3 meters in height. This is most
relevant for hosts in transplant studies and lichen
plots, but should be estimated for studies involving
collection of lichens from multiple host trees.
Accuracy
34
Standards
-
+ 1 meter.
DNSO - Distance to Nearest Small
and lichen plots, but should be estimated for studies
involving collection of lichens from multiple host trees.
Estimate the average distance to the nearest object
that is less than 3 meters in height, such as bushes.
This is most relevant for hosts in transplant studies
DATA MANAGEMENT
AND ANALYSES
- + 1 meter.
Standards
Accuracy
Isebrands, J.G. and S.J. Steele. 1992.
active radiation (PAR) indicator.
1992 Activities Plan.
Photosynthetically
USDA
data sheet or
of quatifying the site variables should be
constructed with subsequent data analyses in mind.
the method
It may be desirable
to construct continuous
Review.
the lichen
percent canopy opening
species),
regressions
cover), principal component
The quantification
analyses.
analyses (e.g.,
Flechten in Beziehung
and lichen
analyses, and covariate
In
140. Jena.
of site factors will aid
Pollution.
in mapping lichen distribution and abundance using
or geographic information systems
Wiksells
mapping.
Levitt,
Skye, E. 1968. Lichens
study of cryptogamic
Boktryckeri
1980a.
temperature stress.
monitoring
global change.
Conservation,
networks for
Environmental
Noel,
II: Water, radiation, salt, and other
Academic
Press, San
607 pps.
D,
Performance
to
a
environment.
pps. 238-282.
Environment,
Atmospheric
23(3):603-609.
Ferry, B.W., M.S. Baddeley and D.L. Hawksworth
and Lichens.
Oosting, H.J. 1956. The Study of Plant Communities.
An introduction to plant ecology. Second Edition.
W.H. Freeman and Company, San Francisco, U.S.A.
The
of London, Great Britian.
Press, University
Academic
497 pps.
J.J. Hechler, and H. Roberge. 1989.
of sulfation and nitration plates used
monitor atmospheric pollutant deposition in
real
1973. Lichen physiology: Progress and
In Ferry, B.W., M.S. Baddeley and D.L.
Hawksworth (eds). 1973. Air Pollution and
Lichens. The Athlone Press, University of London,
(eds).
Athlone
U.S.A.
Second Edition.
Francisco, U.S.A.
pitfalls.
Air Pollution
Chilling, freezing, and high
Responses of Plants to Environmental
Volume
stresses.
Farrar, J.F.
1973.
1980b.
Stresses.
18(2): 173-175.
Great Britain.
J.
Levitt,
Design of ecological
Air
region. Almqvist and
AB, Uppsala, Sweden.
74.
Second Edition.
Press, San Francisco,
REFERENCES
1991.
and
epiphytes and
Responses of Plants to Environmental
Volume
Stresses.
R.G.
der
Flora
zu ihrer Verbreitung.
environment in the Stockholm
conventional
Bailey,
Range,
1992.
Lange, O.L. 1953. Hitze- und Trockenresistenz
and number of lichen
(e.g., elevation
In
Chapter 1.11, pps. 1-12.
Air, Watershed and Ecology (RAWE) Staff Group,
USDA Forest Service, Northern Region, Missoula,
MT. (ECODATA).
species.
This can be done by correlation
Section
Protection
Park, NC 27709.
Jensen, M., W. Hann, and R.E. Keane.
variables
to reflect site factors to relate them to the condition
of
Forest Service and Environmental
Agency, Research Triangle
possible,
II,
Whenever
I:
data recorder.
A
coded for entry first onto a hard-copy
into a personal
In Forest Health Monitoring,
mentioned above be
J.
We suggest the variables
p.
Field 106
Object
389 pps.
440 pps.
Prussia, C.M. and K.T. Killingbeck.
Hisham. 1992. A passive
sampler for atmospheric ozone. Journal of the Air
Grosjean, D. and M.W.M.
and Waste Management
Association,
Holdorf, H. and J. Donahue.
1990.
Concentration
foliose lichens: leachability,
42(2): 169-173.
#15, Montana
Experiment
Station, School of Forestry, University
seasonality, and the
influence of rock and tree bark substrates.
Miscellaneous
Publication
1991.
of ten elements in tow common
Lichenologist
Forest and Conservation
Skye, E. 1968. Lichens
of Montana, Missoula.
and
Air
Pollution.
cryptogamic epiphytes and environment
Stockholm
Boktryckeri
35
The
94(2):135-142.
A
study of
in the
Almqvist and Wiksells
AB, Uppsala, Sweden. 123 pps.
region.
Chapter
PART
1
-
3
APPENDIX
GENERAL DATA FORM
MANAGEMENT DATA FORM
Management Unit Data Sheet
A.
B.
Heading
1.
Unit name
2.
Managing
agency
Geographic location
1.
State
2.
County
3.
Land management
unit
Nearby
large city 1 (distance and direction)
Nearby
large city 2 (distance and direction)
Nearby topographic
features (Mt.
Ranges)
6.
USGS Quad name, 7.5' or
7.
Approximate boundary of unit by Latitude and Longitude, or Universal Transverse
Meridian (UTM), or Township range and section; and description
appropriate)
(if
15'
Physical environmental factors
1.
material (general)
Geology/parent
2.
C.
Soils (short description
or soils map)
36
Landform(s)
4.
5.
(general)
a.
Terrestrial
b.
Aquatic
General
cliamatic factors.
a.
Annual
b.
Length of growing season
c.
Range of elevations
d.
Other factors
Specify the following climatic factors.
precipitation (10-yr mean)
Air chemistry
a.
Types of known pollution sources within 100 km of study area.
Specify all known sources of air pollution and approximate distance to
study area.
Source 1:
Location 1:
Source 2:
Location 2:
Source 3:
Location 3:
Source 4:
Location 4:
Source 5:
Location 5:
Type and Location of nearest continuous
Type of Monitor 1:
Location of Monitor 1:
Type of Monitor 2:
Location of Monitor 2:
Type of Monitor 3:
Location of Monitor 3:
Type of Monitor 4:
Location of Monitor 4:
D.
Biological Factors
1.
Biome type(s)
37
or passive air quality monitoring
station.
2.
Unique
E.
Significant
disturbance
a.
Natural:
b.
Anthropogenic:
history
animals:
Any other Unique Features
38
PART 2 - SITE-SPECIFIC DATA FORM
PRINCIPAL INVESTIGATOR DATA FORM
Field
Key ID,
1 Ag
_ _
__
2 St
__
3 NF
__
Yr
4
1-8
5 Pit
___
_ _
6 Mo
7 Day
8 Name
Sample
Sample
Forms:
9
10
14 Unit
15 Permanent
PRI
17 Comparison
16
18 Potential
Vegetation
Vegetation
12
_
_
Plot ID
Plot ID
Form
Strata:
19
_
LF
Classification,
20 LSC
_
21
_-___
27 LL Dom Sp1
22 CC
DSC
_
SpFea
28 LL Dom Sp2
._...
/
38
41 Aspect
VerPS
+
BS
.
+
Gr
55 Down
33-35 PMat
/
_
39
HorPS
_
40 Elevation
+
Ro
+
+
LD
Log Dia
+
+
Wo
53
BV
ML
56 Dom. Layer
39
_
44 EroType
43 EroStat
42 Slope
45-52 GCov
Field 29-56
/
30-32 Landform
36-37 Pos
Sp2___
26 ML Dom
Site Data,
FD
Field 19-28
24 UL Dom Sp2
25 ML Dom Sp1
54
13
Layers:
23 UL Dom Sp1
29
Field 9-18
11
Existing Vegetation
Existing Vegetation
Data,
System
Ht.
Wa
FLC
Data,
Vegetation
Live Tree:
_
57 DBH
Tree Cover:
61
Tot
Dead Trees:
58 Ht.
_
Field 57-75
62 Seed
Herb Cover:
Cover
Animal
Evidence:
Disturbance:
_
77 F
76 M
79 (1)
_
71 Tal
75 Moss
Field 76-84
_
78 A
80 (2)
82(4)
83 (5)
__
67 VLarge
74 Fern
Disturbance Data,
Ground
64 Pole
70 Mid
73 Forb
72 Gram
60 Ht.
66 Large
69 Low
68 Tot
_
63 Sap
65 Med
Shrub Cover:
59 DBH
81 (3)
_ _
84 (6)
Microsite/Host Data, Fields 85-106
Crown/Bole
87 GH
90
85 LCR
Features:
/
BHL (all)
91
98 BM (1)
Bark Color:
101
Host Tree Location:
89 BAL
88 BTL
BHL (1)
94 BCHL (all)
BM:
86 CD
92
/
BHL (2)
95 BCHL (1)
/
/
BC (1)
104
/
/
93 BHL (3)
96 BCHL (2)
99 BM (2)
/
/
97 BCHL (3)
100
102 BC (2)
HTL
105
DNLO
Comments/Code Explanations
2,
3,
4,
5.
6.
40
BM (3)
103
106
1,
/
/
DNSO
BC (3)
/
_.
/_
Species and Communities
Cliff Smith, Linda Geiser, Larry Gough, Bruce McCune,
Bruce Ryan, and Ray Showman
This chapter recommends sampling techniques for air pollution studies
for lichen species and community monitoring. The
techniques evaluate the condition of and changes in lichen vegetation, use
lichens to estimate air pollution concentrations, and measure responses of
lichens to air pollutants in a manner that will meet quality assurance (QA)
that are appropriate
and quality control
(QC) requirements established by federal agencies.
Some options are provided, along with guidelines for making necessary
choices.
We discuss design principles
that
in conducting
must be considered
or monitoring program using lichens to evaluate air
pollution. Three sections follow on the recommended approaches: general
lichen survey, indicator species, and quantitative community surveys. The
general lichen survey is a relatively simple and inexpensive biomonitoring
technique but is best used around point sources. It is not useful for
monitoring small changes in pollution or where relocation of study sites is
difficult. The use of lichen indicator species is an effective, sensitive, and
any sampling
means to monitor lichen responses to air pollutants.
inexpensive
Unfortunately,
pollution responses of only
a few species are
Quantitative community surveys provide detailed
of many species. The techniques can be time-consuming,
We conclude with some recommendations
DESIGN PRINCIPLES
monitoring
and time constraints
excuses for not conducting
a well-designed
element balances, factors analysis,
ratios to identify
program. In most instances, projects carried out
atmospheric
without adequate attention to QA/QC are not worth
the time, money and effort spent. We recommend
Source-based
inaccuracies
three different
dispersion
approaches
that can be fine-tuned
to
evaluate air quality using lichens. If the requirements
of these approaches cannot be met, obtain
of
in the emission
inventories
models, whereas receptor-based
See
Chapter
Chemical
(see
models
Sensitive
Species,
and Chapter
Analysis, for details on monitoring
and
analysis techniques.
There are two opposite approaches to assessing
Several hypothetical
models are shown
along with possible explanations
emission
point source (Gough
and dispersion
predictions,
of an element in
while
41
a
and monitoring air pollutants: source-based models
and receptor-based models. Source-based models use
inventories
or
and
a
the authors of this
for consultation
sources.
models suffer from uncertainties
for
in figure
the concentration
receptor versus distance from
et al.
1986).
1
Appendix
assistance. Many
are available
and natural
suffer from the difficulties inherent in identifying
contributions from multiple sources at
receptor site.
I).
handbook
anthropogenic
elemental emission
a
professional
element
trends, and stable isotope
concentration-distance
research
factors,
models use enrichment
receptor-based
are often cited as
approaches
programs.
for reporting results.
chemical
Funding
All
require highly trained personnel.
7,
and generally
can be used for a single sample or for long-term
5,
evaluations
expensive,
the air
known.
adequately
Model I
INFLUENCE
ZONE
Model IV
BACKGROUND
i
A
O
0
DISTANCE
A
DISTANCE
II
J_
0
A
DISTANCE
0
A
DISTANCE
m
j_
O
A
DISTANCE
Figure 1.
- Hypothetical
receptor-based models of the concentration (C) of an element
versus distance (D) from a point source contamination.
In Model I, a point source emits an element M.
receptor sites are sampled at one instant in time
same elemental species (e.g. fine particulates)
All
and are assumed to be equally
In this model there is
of M
concentration
efficient
a negative
the same point.
with
In Model IV,
the
the background
Model II illustrates
of
a situation
the emitted
in which chemical
the reaction product.
situation
might be oxidation
III
variations
An example of this
of an emitted species
of M
on dispersion
if
varies greatly.
in the regions
of
influence
diurnal,
or
or on the receptors,
influence
source can be expected. Differences
around a point
in the regions of
might be obtained based on the selection of
soil versus vegetation
or sampling of different plant species.
the receptor used, such as
similar situation to
model I; however, two point sources, or pseudo-point
sources with different regions of influence, emit the
demonstrates
concentration
seasonal influences
during transport.
Model
negative correlation
and distance may be found
vary by the source, or there are climatic,
species occurs during
transport away from the source, and the receptor only
collects
be
Model V demonstrates the influence and
importance of the sampling time. If the emissions
is reached. This distance (O to A)
of influence for the emitting source.
defines a region
conversion
a less distinct
between concentration
versus the distance from the
source, up to the point at which the background
concentration
from
An example of this model would
a point source situated on the ocean coastline.
collectors.
correlation
in a receptor
sampling,
a
Hence, regions
of
influence
determined
technique should only be considered
42
by this
as the minimum
region of influence,
and caution
change laboratories
should be used in
results where competing
interpreting
emission
work,
may be involved.
Contamination
of
See
data.
Chapter
be measured.
Gradient
10
Principles
reviewed by anyone beginning
study. They are paraphrased
What is "Baseline"?
1.
differ.
which ideally exclude human
concentrations,
A
baseline
2.
represents the concentration
measured at some point in time, a "snap shot", and is
typically not true background. A background
represents an idealized
situation
difficult to measure than
A
true background
and analyzing
and is typically
more
a baseline.
are best defined
ice cores, or varves
as a range
transformation
the geometric
the geometric
to normalize
of values
4.
To test whether a condition has an effect,
collect samples both where the condition
is present and where the condition is
absent but all else is the same.
5.
Carry out some preliminary sampling to
provide a basis for evaluation of sampling
design and statistical analysis options.
6.
Verify that your sampling device or
method is actually sampling the
population of interest, and with equal and
adequate efficiency over the entire range
of sampling conditions to be encountered.
data, the mean becomes
mean (GM) and the deviation becomes
deviation (GD). The 95% expected
range is the range defined by the GM/GD
to the
GMxGD2 (Tidball and Ebens 1976), where GM and
GD are calculated from data generated by nested
ANOVA
designs (see Chapter 7).
"working" baselines can be approximated
So-called
when examining
the distance
vs. concentration
data
generated by studies using transects that radiate away
from a contamination
point source. A well-defined
trend will show an inflection point, the asymptote of
which can be used
as a
7.
working baseline.
Stability Through Time
of
8.
study designs and statistical methods is
Jackson and Gough 1991).
of field studies and laboratory
procedure should be of major concern in the planning
of a study. Identical field and laboratory procedures
lacking
(see, for example,
Repeatability
must be maintained
If the area to be sampled has a large-scale
environmental pattern, break the area up
into relatively homogeneous subareas and
allocate samples to each in proportion to
the size
Investigations of temporal variability in air quality
studies arc few, and a thorough understanding of the
proper use
Take replicate samples within each
combination of time, location, and any
other control variable. Differences among
variables can only be demonstrated by
comparison to differences within.
number of randomly
replicate samples for each
combination of controlled variables.
95% expected range. Following log
representing
you
allocated
from lake or ocean sediments. Biogeochemical
baselines
Be sure to state concisely the question
3. Take an equal
may be obtained by sampling
tree rings, glacial
an air quality-related
here.
are asking.
is intended to represent natural
Background
influence.
and "background"
vary, the
R.H. Green (1979) presents 10 principles of study
design and data interpretation that should be
values.
The terms "baseline"
procedures
This is not an easy task.
Green's
These estimates help establish baseline element
concentration
If
error associated with this variability must somehow
Analysis. Over the last
fifteen years, a protocol has evolved involving both
gridded and non-gridded study designs which help
define the sources of variability in biogeochemical
5
for a discussion
are unknown.
(2)
the field
(3) sample at the same time of the year under
and (4) do not change
sample handling and storage.
spatial and temporal trends, and
from some norm
deviation
procedures,
performing
similar weather conditions,
gradients are often suspected but
their magnitude,
or analytical
avoid changes in personnel
sources
of
the subarea.
Verify that your sample unit size is
appropriate to the size, density, and
spatial distribution of the organisms you
are sampling.
whether the
error variation is homogeneous, normally
distributed, and independent of the mean.
9. Test your data to determine
through time. Some things to
watch out for: (1) once a study has begun, do not
43
10.
Plots on Bark or Wood
Having chosen the best statistical method
to test your hypothesis, stick with the
On living trees, permanent markers should be
damage and defacement of the
bark. Aluminum nails and tags are preferable to other
kinds of metal markers.
chosen to minimize
result.
Quality Assurance/Quality Control
Plots on Rock
The corners or endpoints
of quality assurance and quality control
(QA/QC) will vary between studies depending on the
objectives of the study and the available funding.
The level
While minimum levels
rigorous
are outlined
levels are provided
more rigorous
approaches
here,
objectives
are appropriate
and the potential
should guide in the choice
uses
of
if
study
The study
the study results
of QA/QC level within
25 percent to the total budget.
re-located.
is an important
of
of
of
a quadrat can be
the corners can be
plot corners may be necessary;
Repainting
the plots are measured.
voucher
Alternatives
aspect of all studies, even
generate key reference material for later studies,
accuracy
and consistency
other information
between studies.
In some cases the specimens may be required
evidence for litigation purposes.
The collection, storage, organization and
to rocks with
as
of
specific
sample
it may not be
or one corner
a transect
An appendix of
marking
final report should be
the
produced for each study. This appendix
should
include the actual data used in the analysis,
used in the analysis,
photographs
difficult to re-locate during follow-up studies. Global
Positioning System technology is becoming cheap
any
and any maps
information relevant to the
inclusion of this material generates an
and/or other site location
study.
can be
If
the
undesirably
minimized.
published
can be
long report, the appendix
separately from the report and fewer copies
produced, perhaps in a less expensive
Metal Nails or Tags
format. These
in the distribution
can then be included
These should be located so that water runoff does
not pass through
of
the other points
Archiving Data
but they are more
problems
(with
plots.
or desirable to use conspicuous
permanent markers. Plots located far away from trails
or campgrounds are less likely to be disturbed, and
enough that most marker relocation
a plot
Monitoring, for precautions on permanently
permissible
also less subject to vandalism,
to
being located using a
compass and tape). Sec Chapter 6, Active
several quadrats or transects
wildernesses,
or nailed to nearby trees.
or trees to mark one end
in close proximity of each other. In some
study areas, especially
scratched into them, can be affixed
silicone
Although stakes can driven into the ground
Permanent Markers
if
paint dots, such as
mark plots, a better method is to place tags on rocks
of
Re-location and re-sampling
of
Plots on Soil or Moss on the Ground
availability of voucher specimens is discussed fully
in Chapter 2, Floristics.
units are facilitated
to the use
drilling holes into the substrate, may be difficult.
Aluminum tags, with the sample identification and
those which are not purely floristic. These activities
are located
at least two
an adequate paint supply should be taken each time
The collection and preservation
promoting
if
reconstructed
Vouchers
specimens
plots on rock should
permanent, although the outline
In general, QA/QC requirements
budget constraints.
add an additional
more
in EPA manuals. The
results could be involved in litigation.
of
with small daubs of red or
orange paint which are clearly visible but
unobtrusive. Car touch-up paint in a small tube with
a brush attached to the inside of the lid is
convenient, as is nail polish. The paint will stick
better if it is placed in a slight indentation in the
rock made with a hard nail. On crumbly or
exfoliating surfaces, daubs of paint may not be
be marked on the rock
of
the report
This approach assures that
multiple copies of this material will be in circulation,
promoting the availability and future accessibility of
only when appropriate.
the sample unit and affect the
results.
this reference material. The second archive
is a "local"
location,
administrative
44
location
that is the office or
location
of
relevance
to the study site.
which can serve
The third is a central location,
of
The following types
reference material should be
it was actually
of
publication
equipment
or
Any
the methods should be provided.
field notes or research management notes.
Finally, raw data in any form (field notes, lab
sheets, photos, etc.) should be available in sufficient
the
5) site location information which may
include maps;
detail to enable a researcher to trace back to the
6) photographs.
should also be annotated with any observations
observation
original
Not all of these types of reference material
necessarily exist in all studies. These reference
Archive
The general lichen survey
locations.
Local
Appendix
etc.
General
maps
Specific
site location
X
X
information
X
X
X
also track changes in air quality
X
X
scale of increasing
X
X
X
X
X
over time. On
structure and complexity,
the
general survey bridges the gap between the floristic
method
of
complex
X
(1988,
Wetmore
quantitative
1990) and the more
methods of Case (1980),
McCune (1988), Will-Wolf (1980), and others.
The survey method
similar to the classical
is
Photographs
simple
over fairly large areas. Repeat studies can
differences
X
relatively
to indicate air quality
method designed
biomonitoring
Central
a
—
Raw data (database)
data (database)
of
archive locations
a
1.
Analysis
that
later analysis
GENERAL LICHEN SURVEY
Reference material
Field notes, lab sheets,
interest. These data
the data.
1.
Table
regarding
is
in Table
of
any datum
might have implications
materials and their recommended
a
or
document
from these methods should be annotated in
deviation
etc.);
are listed
of
an explicit description of methods
Secondly,
reference to an easily accessible
maps (geological, topographic,
4) general
of
(names and addresses), descriptions
used, and so forth.
2) raw data such as the data initially entered
in a computer database;
as
contact persons who are knowledgeable
some aspect of the study, a schedule
research activities, site locations, field personnel
actual field notes, lab sheets, etc.;
3) analysis data, the data
used in the analysis;
of
to) names
concerning
in this discussion:
included
1)
Documentation should contain general
background information including (but not limited
as a
reference center for this type of material.
nation-wide
In this system, the appendix serves to make
readily available
After
the
study, the appendix
to
initial
The central location
of
serve as
repository
of all
condition
activities
throughout
to
a
is
Connecticut.
The survey method can be used to study lichens
with
reconnaissance
the least
availability
eastern U.S.,
wide range of
advisable
and distribution
generally
to determine
in
of air
study sites are chosen to contain
45
substrate
study area. In the
accepted that corticolous
lichens are the best indicators
this manual.
a
forward
also applicable
discussed
of
on any substrate. Usually some preliminary
down time or methods errors. Systematic
documentation
over the entire state
is
it
a
study and move
study
the
Description of Method
methods, systematic
of work to date, and proper archiving
reference material will enable other researchers to
pick up
of
(Showman 1981, 1990). Metzler (1980) used the
survey method to map species distribution and
Program Continuity
The consistent use of
of
(1973, 1975) to
around coal-burning power plants
by Showman
sites can also document changes in air quality
relevant to that management unit.
documentation
the
in the Ohio Valley region. Re-surveys
as
studies and/or sites.
a
number
will
The local location
the material
will serve
In
indicate air quality
to anyone interested in obtaining
from
a
a
by request from both the central
(see Ferry et al. 1973 for good
United States, this method has been
workers
used extensively
should be available
clearinghouse
examples).
and report
and local locations.
information
European
it
is
of
a
anyone who might request
distribution
it.
general study reference material
species mapping approach used by numerous
the
quality.
Thus
several to many
unshaded trees. Sites can be further standardized
tree
between healthy thalli and thalli with obvious
to
species or bark type, but this may severely limit
of
the number
available
utilized hardwood
trees in open situations
and cemeteries.
churchyards
contain
sites. Showman
3-6 trees and occupy
of
(1975)
such as
substrate should be standardized
possible.
A
selection
criteria.
careful
Species richness (number
of air quality
0.1-0.2
The number of study sites used in
depend on the size
of
will
the study area and the desired
This approximates
square kilometers
(5 square miles).
by the general survey method.
between forested areas
deposition
Richness
of
Pennsylvania
(Long 1990).
average about 10 sites per 7-1/2 minute topographic
map quadrangle.
collected
in
within a study area. Data can also be
summarized in terms of total richness and mean
richness per site for comparison between study areas.
Showman and Long found species richness to be a
simple yet sensitive indicator of differences in
density. Biomonitoring studies by Showman
sampling
Richness
distribution
site
a survey
species) is also a
(Showman 1988).
values can be easily obtained from data
values can be mapped to detect anomalies
to whatever extent
of
of
good indicator
rock or soil
record should be kept
data may be presented on the same
map.
hectares (0.25-0.5 acres). In some regions trees may
be absent, and saxicolous or terricolous lichens must
be studied instead. Sites containing
of
different kinds
These sites typically
an area
By using different symbols, several
necrosis.
one site per 13
A
large study
Discussion
area with a low pollution gradient could be covered
by a lower site density, while a small area with a
The general lichen survey method is applicable
of the study
of air quality.
steep pollution gradient would require a greater site
where the purpose
is to determine large
density.
overall patterns
The method is very
For
valid temporal study, it is necessary
a
relocate the same study sites precisely
In populated areas, this can be accomplished
accurately
marking
complex
on a 7-1/2
The primary
at each site should
the date, site identification
using
ECODATA
advantage
simple and fast.
number, and the
as
for
of
this method is that it is
Site standardization
the quantitative
forms. The lichen
of
to be covered at a moderate cost.
presentation are also relatively
condition,
done on a day-to-day
i.e., bleached,
discolored
15-25 sites per day
a larger area
allowing
species present are then recorded. Notes on thallus
or fragmented,
fertility, and size may also be recorded.
is not as critical
methods so a broader range of
sites can be used. A survey
can usually be accomplished,
study area description. See Chapter 3, Site
Characterization,
for more details and for
characterization
is difficult.
areas.
permanent marker may be necessary.
include
where site
This is a "quick and
dirty" method best suited for use in areas with
medium to high pollution levels and in populated
map. However,
The general data recorded
terrain and ecology
standardization
by
in remote areas
may be more difficult, and some
minute topographic
site relocation
the site location
appropriate around point sources and in areas of
to
at a later time.
Data analysis and
simple and can be
basis.
The primary limitation
of
the survey method is
that it is not as sensitive to small changes in air
quality as the quantitative
Data Analysis
Data from the general lichen survey are usually
presented cartographically.
mapping are discussed
methods are.
less appropriate for remote wilderness
non-vehicular
Some general aspects of
travel and difficulties in site
location/relocation
by Showman
It is also
areas where
negate the primary
advantage of
this method.
(1988).
Distribution maps are prepared for the common
species and then compared to known or suspected
pollution gradients. A discrete void in the distribution
of one or more species downwind from a pollution
source could indicate an air quality effect. A random
pattern of absences would suggest some other factor,
such as differences in site ecology. Species
distribution maps might also include other
information such as injury or fertility. Metzler (1980)
presented distribution maps which distinguished
INDICATOR SPECIES
The indicator
species method can be used to assess
changes in growth or other parameters in one or
more known air pollution-sensitive
quantitative
46
methods.
lichens using
loss of this species?",
The indicator species approach is useful when
target species are known
particular
to occur in the
study area.
Such species are usually
information
on pollution sensitivity
little
ones for which
is available
suitable for using the species as monitors.
If
in
and reasonably
Species with distinctive
with
strongly
desirable,
are especially
monitoring
the background
to particular
since this is analogous
are below the detection
concentrations
colors that contrast
Choice of Indicator Species and Response
Variables
species that
pollutants,
Determination
involves
of suitable indicator species
of many features, as discussed in
evaluation
more detail in Chapter 5, Sensitive
number of indicator
study area to determine which lichen taxa are
can make a preliminary
present, the investigator
species are
assessment of likely indicator
is required to
identified,
only minimal
recognize
them in the field and follow changes in
their growth and condition over time.
Variables to measure in determining the response
of indicators include cover, elemental or other
Chapter
5.
Sample
Units
of permanent sample units requires
Selection
species approach
knowledge
to a particular
The growth form of the thallus affects whether
photographic
or other methods of determining
changes are used.
Growth forms should
in choosing
considered
study area.
specific
measuring or estimating
parameters.
Many of
approach is that it gives information
on only a few
species and says nothing about the others. In
or to the
presenting the results to decision-makers
the lichen
sensitive to pollutants
indicators
methods
growth
be
of
cover or other growth
species which are known to be
and are the most useful as
have a fruticose
or foliose thallus.
The
growth of such taxa may be difficult to measure by
there is a danger of the "snail darter
syndrome",
the substrate, and other factors.
Lichen Growth Forms
More data are needed on the sensitivity of species.
Another apparent limitation of the indicator species
public,
of
topography
caution must be used in
that information
number and size
(see discussion of quantitative community sampling
later in this chapter). The choice of plot type
depends on growth form of the lichen, kind and
of which species are
sensitive. This is a problem in most areas of the
United States, especially in the West, where the
pollution responses of few, if any, species are
Even
known.
See Chapter 5 for more information.
when some data on sensitivity are available for a
applying
the
number, size and
to consider
arrangement, and tradeoffs between
Limitations
in
content, and others as discussed
chemical
investigator
species, considerable
partner, and on
from the literature when available.
information
The indicator
than the general survey method.
the other hand, the indicator
species, based on
growth form and photosynthetic
species method gives efficient focus on the air
pollution problem. If high-tech methods are not
applied, the indicator species approach is
inexpensive, involving little time on the part of
personnel, and while it does not require complex
analyses, it is more sensitive to changes in air quality
On
Species
after walking through the
However,
Responses.
Advantages
presupposes
the
if video
tolerant may also be useful: an increase in their
abundance can indicate increased pollution.
training
limit of
instrument.
are
Once a limited
to using
that air pollutant
an instrument and reporting
and with other lichens
particularly
of any lichen.
is the only
results can still provide
such "negative"
reliable conclusions,
species are usually ones likely
Although indicator
to be sensitive
However,
and
are used.
techniques
significance
species monitoring
species chosen may not show any detectable changes
to the influence of air pollution.
over a large portion of the study
occur frequently
area.
rapid growth rates,
or ecological
indicator
when in most cases
that can be ascribed
the
field, have individual thalli with fairly discrete
boundaries
especially
is known about the potential
approach used, there is the risk that the particular
The most
easy to identify
suitable species are relatively
anything
economic
from
studies, and ones with other characteristics
previous
if
i.e., people may ask "who cares about
photographic
means because
form of the thallus.
lichens, especially
47
of
Pendulous
the
three-dimensional
(beardlike,
hanging)
those hanging from branches and
Lichens on Soil or Moss
mostly free from the substrate (e.g., many Bryoria
and Usnea species), present a different
from other forms,
problems
set
of
Some of the most abundant and potentially
since growth is primarily
in length, rather than in area or diameter.
growth
of
indicator
The
large, loosely
included
of
partly
be found
in other types
of
for doing so can
studies on lichens,
and in
studies on vascular plants.
small, scattered subunits (areoles
and species with the thallus mostly or
Methods of Using Plots
in the substrate are usually very
difficult to follow. Often frequency
(presence/absence) is the only parameter that can be
measured for such species. Crustose and appressed
foliose lichens, forming flat, solid patches or rosettes,
with growth entirely in the radial direction, are the
immersed
easiest ones to measure by photographic
methods
studies related to
but the methodology
numerous
or squamules),
entirely
in permanent monitoring
air pollution,
in
Such species have rarely been
alpine or desert areas.
foliose lichens (e.g., Lobaria spp.) consists of
increases in both diameter and thickness.
Growth changes in lichens composed at least
useful
and
occur on soil or moss, especially
rapid growing)
(shrublike) fruticose lichens, and
attached and suberect or subpendulous
caespitose
easily identified,
lichens (sensitive,
of
determining
cover, but few
species are known to be sensitive
Photographic Methods
Photographic methods are most useful for
lichens
following changes in two-dimensional
(crustose and most foliose forms) growing on
two-dimensional
or other
of
surfaces, especially
than 90 degrees from horizontal.
these
in surface features
to air pollution.
Photographing
overhanging
less
the substrate, the more
and focus problems
depth-of-field
Another potential problem with crustose species is
the difficulty of obtaining voucher specimens, which
must be collected with pieces of the substrate.
of
at angles
The more variation
quadrats, especially
will
occur.
on steep or
surfaces, or in partial shade, is
process, involving two
sometimes a rather awkward
people. One person holds the frame in place or holds
Substrate and Habitat Considerations
up something to give uniform lighting or shading,
while the other person takes the pictures.
For easier identification of species and thalli in the
photographs, it is helpful to restrict the number of
species within the sample unit to one or a few kinds,
preferably ones with distinct colors that contrast with
each other and with the background. Video methods
with digitizing capabilities are very useful both in
enhancing contrast and in determining the area
covered by the thalli, especially species composed of
Lichens on Trees or Shrubs
At many sites, the majority of species growing on
bark or wood (including several of the most sensitive
lichens)
on the branches and in
are abundant mainly
the canopy
rather than on the boles
of
trees, and are
from the ground. These situations
require special methods, including climbing the tree,
not accessible
or special plotless
methods to estimate abundance.
Likewise, shrubs may be
lichens,
requiring
a
major substrate for
development
of
scattered units, but such methods may be
prohibitively
special techniques.
Lichens on Rock
It is difficult to follow long-term
growth
of
lichens
Cover and Frequency Determinations
Measurements or estimates of cover and frequency
changes in the
found on exfoliating
can be done in any situation,
or unstable
surfaces, or in areas likely to be covered by litter or
overgrown
by other plants.
Lichens
the lichen
on small or
avoided.
Many sensitive lichens preferentially
including
video techniques,
determinations.
grow
including cases where
are three-dimensional.
with photography,
in making the
In cases where lichens do not
contrast much with the substrate or with other
surfaces, or in crevices and
shaded areas where photography
species as in shaded areas, cover determinations
and other methods
difficult to use.
Although aquatic or semi-aquatic
species may be potentially useful monitors of
acidification, it is difficult to study them and to
establish permanent quadrats or transects for them.
are
or surface or both
They can also be combined
unstable rocks likely to roll down hill should be
on steep or overhanging
expensive.
may
require use of a hand lens or even a flashlight to see
the boundaries
48
of
the
thalli more clearly.
Line-Intercept Transects
Tape Methods
Ryan and Nash (1991a) used
transect tape to determine
Various non-photographic
quadrat methods have
also been used in many studies. Ryan and Nash
a 20 m long
of
cover and frequency
lichens on rock, for community
(1991b) located quadrats randomly along a transect
line across a field of small boulders and estimated
flexible
percent cover within each quadrat. Such methods
analyses. Though
may be useful where it is desirable to make a
relatively rapid survey of a large area with numerous
their transects were not designed to be permanent,
the same method could
be adapted for permanent
transects focusing on one or a few indicator
replications.
This method is generally more difficult and less
reliable than quadrats or very short transects, and is
probably most useful in studying lichens growing on
very large, contiguous patches of rock or soil.
Denison (1990) sampled lichens on branches by
wrapping a flexible tape in a spiral along the length
of the branch. Such an approach may be useful in
some cases, but not for making permanent plots,
because even on a tree trunk it is difficult to place
the tape in exactly the same place each time the
sampling
However,
changes in
when monitoring
species over time, it is desirable
indicator
species.
repeatability
of
the cover
estimates by using a grid of
squares or pins within the quadrats.
Plotless Methods
Examples
of
several plotless
methods for
estimating the abundance of lichens
are given by
Ryan and Nash (1991b). In that study, the individual
sampling units were not permanent, since they were
selected so that it is impossible to return to the same
is done.
trees or rocks to record changes. Plotless
Pin Intercept
Methods
generally do not produce the precision
For lichens growing on trees with large, straight
trunks, the "dotiometer", a moveable pin attached to
a meter stick, may be useful. This method is
discussed by Ryan (1990) and is described in the
1988 U.S. Forest Service Region 5 draft protocol. If
methods, but in some situations,
they are the quickest
so that the transect
are strongly
the advantage
lichens are
of
trees,
and most efficient way
of
approach.
Analysis of elemental or other chemical content
methods. This subject is
discussed in detail in Chapter 7, Chemical Analyses.
can be done with plotless
of being easier to
three-dimensional.
However,
in most
Data Analysis—Indicator Species
if
The type of data analysis
sampling
Quadrats
Data Storage
if
and Retrieval
Data should be stored in an appropriate relational
database, as described by Looney and James (1990).
Files
1)
on tree trunks, but also on rock and soil. Video
be useful
to be analyzed.
reported establishment
and James (1990) using a wire grid, is recommended
for most situations, and should work well not only
with digitizing may also
financially feasible.
needed depends on the
designs and methods used and the kinds of
Although various studies have
of permanent plots to follow
indicator species over time, few have presented
analyses of the results.
data
Various methods of establishing permanent
photographic quadrats for sampling lichens on rock
have been used in a large number of studies,
including the "quadpod" described in the 1988 U.S.
Forest Service Region 5 draft protocol and used by
Ryan (1990). Other studies (e.g., Skorepa and Vitt
1976) have used various types of photographic
quadrats for lichens on trees. The method of Looney
are
branches
plot
where the substrate, lichens, or both,
situations a quadrat method may work as well as,
not better than, pin methods.
techniques
as where
methods
Computerized video techniques may allow the use
of entire trees or rocks or parts of them as permanent
sampling units, which could be considered a plotless
that can be moved up and down may also be used.
Pin methods have
of
sampling.
will not be greatly affected by changes in the
diameter of the tree. Frames with fixed rows of pins
use in situations
mainly on inaccessible
growing
such a method is used, it is important that the meter
stick is oriented nearly vertical,
to make
such quadrats permanent and to increase the
to be established
include the following:
name and code,
Sample unit information:
location, date, notes, etc. for each unit;
2) Cover and frequency data, coded to site,
sampling unit, and year;
they
3) Photographic records;
49
4) Maps and aerial
Looney and James (1990) recommended using
percent cover data to calculate relative growth rate
photographs;
of
5) Locations and collection numbers
voucher specimens, and
(RGR), which allows initial size differences to be
and the net changes or performance of
discounted
individuals
6) Information on species observed at each
site other than the target species.
Some response variables,
semi-quantitative
such as physical
Detailed
same or different
format is described
techniques currently
of
the
experimental
information
to be recorded.
patterns
plot
specific
sampling
community
techniques,
analysis
Community
as
well
as
pollution.
for
of
form for describing
describing
the location
of
reproducibility
of
texts
5 and 7
for
patterns that can be attributed to
is the quality
form.
the need
on any specific
and its response to particular
If
results
the
for further testing of
(e.g., by fumigation,
the sample unit,
and quantity of
about the species, including both its
indicator approach are ambiguous,
the
transplants,
of
the
they may suggest
species'
response
or studies in areas
with known levels of pollution — see Chapter 5),
different method of determining growth or other
parameters, or even choice of a different species.
ones that will facilitate
the
(see Chapters
pollutants or other factors.
reference photos, and comments or
sketches, especially
or
may be useful.
With indicator species, perhaps the most
basic biology/ecology
form should be
on the specific
room should be allowed
parametric
approaches
Standard statistical
important consideration
Analyses).
used and cross-referenced
various
variability.
data available
In addition to forms for specific sampling
Plenty
of
distinguish
(see section on
methods, the general site description
met,
univariate
Interpretation of Data
As with other monitoring approaches, the basic
principles of design must be applied for interpretation
of the results to be valid. The challenge is to
of
as
the appropriate
references).
It may be desirable to
approaches
are
should be consulted
forms for a wide variety of
develop generalized
If
Principal components analyses or other multivariate
techniques are likely to be useful in detecting
3. Here, we suggest
certain types
video
being developed.
design is used and the statistical
non-parametric
Species
ECODATA
the form is not critical
long as the form provides
to
types of
Special
analyses can be used for studying
Regression
this chapter. These are only examples,
and the exact format
also need to be considered
trends over time and space.
and detailed transect measurements.
of some forms that have been used for
of
of
the substrate (e.g.,
Statistics
permanent plots or transects are given in appendices
1 and 2
of
suitable areas
analyses may be required for the computerized
descriptions
Examples
thalli
among thalli of the
understand the patterns observed.
or biological
more generalized
tiny thalli
single-layered
species, or the effects
fissures or depressions)
in the
in Chapter
standard forms for recording
of
distribution
assumptions
- Indicator
such as numerous
or other interactions
competitive
may provide a basis for
site characterization
and
taxa involved.
thalli. Other factors, such as
vs. older multi-layered
lichens.
Standard Forms
situations,
quite different
exist between
to what extent these factors influence
determining
the particular
vs. a few large ones, or young,
visually comparing trends over time or space for
each plot or site. Scatterplots may be useful for
analyzing some kinds of patterns. Map series with
data for a given species can be useful in detecting
changes over time as well as space.
When sufficient data on pollutant concentrations or
other physical-chemical parameters are available,
these data can be compared with cover/frequency
factors. Such correlations
of
For example, several photos may show approximately
(or other quantitative
by graphically and
whether correlations
the structure, growth,
equal cover by a given species, but may represent
scales. In the simplest cases,
data patterns and non-pollution physical
or other cover data for
species requires a solid
dynamics
population
changes in cover and frequency
data to determine
of
understanding
at best be coded by
parameters) can be analyzed
indicator
monitoring
Basic Analyses of Data
condition or fertility, can
to be compared.
Analysis of photographic
sampling.
50
a
QUANTITATIVE COMMUNITY
ANALYSES
3). Although extraneous factors can be removed to
some extent in the analysis phase, training in
multivariate
of quantitative sampling of lichen
communities for air quality monitoring is the
accurate and sensitive detection of changes in lichen
communities through time. In most cases, another
The objective
caused by other
factors.
General
to be met by the same sampling is the
objective
analysis is required to isolate differences
due to air quality from variation
Design Considerations
species. This is often the case when setting up a
In most cases, sampling designs will be
with subsampling at a number of
individual sites within the study area. In some cases,
as discussed below, a single large plot will be used
at each site. Typically, sites or "stands" are
considered to be areas that, at the scale of the
dominant vegetation, are essentially homogeneous in
vegetation, environment, and history. Sample units in
baseline or when there is uncertainty
the subsample
of differences
description
in lichen communities
along a pollution gradient. Methods
from those discussed below should be used
different
for
hierarchical,
somewhat
characterization
a comprehensive
communities
of lichen
in an area.
Quantitative community sampling is useful when
response information is desired for a large number of
about species
or about specific threats to air quality.
However, such sampling is also useful in any other
circumstance involving a point source of pollution.
some modification is requested and/or made to a
point source of pollution, quantitative community
sensitivities
Designing
the site level and
of sample units at
within sites. Site selection will
(e.g., transects along
among several habitats, and
gradients, stratification
standardization
or historical
these studies produce
variation,
information
to restrict extraneous environmental
factors). The goals of subsampling sites
are to (1) determine accurate abundance values at the
site level and (2) characterize the within-site
on abundance and
relevant to other kinds of management
to allow stronger comparisons
among sites
and through time.
involving lichens (e.g., ecosystem nutrient
problems
balances and food for caribou,
Site Selection
deer, and small
Sites should be selected according
mammals).
Advantages
direct monitoring
permanent study sites. The number of sites depends
stations, the opportunity
monitoring
strategy to locate
interest, then ideally use a sampling
Quantitative community data methods are highly
sensitive to changes in air quality. In areas with
exists to produce calibrated
to the general
in the Design
Principles section and in Chapters 5 and 7. To
summarize, one must define the population of
outlined
study design principles
multiple
decisions
depend on study objectives
Quantitative community data will likely produce
other useful spinoffs, such as improving our
understanding of basic lichen ecology. Furthermore,
distribution
or points.
about the
plots (quadrats)
number, size, and arrangements
If
may detect responses to the modification.
sampling
are typically
a study thus requires
of the vegetation and the goals of
(McCune and Lesica 1992). With some
on the complexity
studies, i.e.
the study
estimation of air pollutant concentrations,
sulfur dioxide, based on lichen data
(Ammann et al. 1988, Liebendorfer et al. 1988,
McCune 1988, Nimis et al. 1990).
one can use standard statistical
quantitative
preliminary
particularly
techniques to determine the sample size required to
sampling,
achieve some specified
practice, however,
without good information
made
based on preliminary
In those cases, one can use a rule of
thumb (slightly modified from Tabachnik and Fidell
1989) that for every important controlling factor or
gradient, 20 additional sites are needed. For example,
This class of methods can be more
than other methods. Special
is required for field workers.
Most seasonal, temporary workers will not have the
background needed to quickly develop the required
field expertise. Site standardization is critical for
these methods, since lichen communities are sensitive
to many factors other than air pollution (see Chapter
taxonomic
are often
sampling.
Disadvantages
time-consuming
In
degree of accuracy.
such decisions
to determine
training
community
differences
along a gradient
from a point source on a homogeneous
along a transect would be reasonable.
substrates were involved
differences,
two
and there were elevational
then 60 sites would be desirable
important factors x 20 sites/factor
51
plain, 20 sites
If
= 60
sites).
(3
Table
2. — Tradeoffs
between
Variable
Bias against
Higher.
of visual
integration
Inclusion
of rare to
species
Accuracy of cover data
on common species
Analysis
Analysis
Recommendations
Lower.
are inadvertently
Small sample
Lower.
Cover classes in large sample units
result in broadly classed cover estimates with
less accuracy and precision than that compiled
More accurate and precise
Higher.
estimates for common species.
of vegetation.
Varies by complexity and degree of development
of vegetation.
No consistent difference from
many-and-small.
few-and-large.
by complexity and degree of development
No consistent difference from
Data entry at site level leads directly to
Point data or microplot
Slower.
initial data reduction (by hand, calculator, or
computer) to site-level abundance estimates.
No estimates of within-site variance restricts
to individual sites as sampling units.
Within-site
analysis
sample
The extreme case
The extreme case (point sampling) is most useful
when rare to uncommon species are of little
concern and accurate estimates are desired for a
common species.
In most cases a compromise
variance estimates not possible
units are larger than points.
The most commonly
epiphyte biomass,
Abundance
are possible.
designed sampling
revisit as exactly
see Stevenson
plan. Subsequent samplings
make relocating
should
and the ability
the same sample units.
the permanent
1988).
to biomass
(Hermy
estimates is possible
1969,
data collection for calibration
McCune 1990, Stevenson and Enns
(proportion
or percent of sample units containing
a
given species) should be avoided because frequency,
unlike cover, is highly
dependent on the size of the
sample unit. Because there is usually
standardization
frequency
proven ability to
comparison
in lichen communities among sites
subtle differences in air quality.
detect differences
with relatively
methods,
1992). This conversion is not normally necessary for
studies of lichens as indicators. Frequency measures
on tree trunks. Only a few examples are
of
estimation
to measure it nondestructively
Conversion
(Forman
plots easier.
depend on the substrate (table 3).
Many air-quality studies have used lichen
listed here, selected on the basis
as an
but requires additional
In most cases, evenly spaced (systematic) sample
units are acceptable because they will not produce
results that differ from strictly randomized sampling.
The specific plan for arranging the sample units will
communities
measure,
and Enns (1992).
in Chapters 2
Cover excels
between different
comparability
Most statistical texts advocate random or stratified
random sampling. However, systematic sampling will
usually
used abundance
abundance measure in speed, repeatability,
should follow a carefully
as possible
units
measures are also discussed
and 5 of this document.
Arrangement of Subsample Units
The initial sampling
of larger sample
excluding presence-absence, is percent cover. For a
review of abundance measures used to estimate
The extremes are single large
a stand (e.g. releves or
intermediates
number
unless
Abundance Measures
between number and size of
habitat type plots) vs. many tiny sample units (point
All
data require
site-level-analysis.
Number and Size of Sample Units
sampling).
cover
units.
by using a smaller
is better.
sample units (table 2).
quadrats to characterize
observer
Low.
Unless sample size are very large, most
rare to uncommon species are missed.
(single large plot) is most
useful with extensive inventory methods.
There are tradeoffs
units force the eye to
spots, reducing inadvertent
selectivity in detection of species.
specific
High. Visual integration described above results
in effective "capture" of rare species in the data.
Faster.
options
1992).
Low. Minimal use is made of integrative
capability of eye, forcing the use of very large
sample size to achieve comparable representation
of the community.
Varies
time
and Lesica
High. The use of visual integration over a large
area is an effective tool against the normally high
degree of heterogeneity, even in "homogeneous"
stands.
from many small sample
time
Sampling
units (McCune
Many-and-small
There is a hazard that some species,
particularly cryptic species,
missed by the eye.
uncommon
sample
Few-and-large
cryptic
species
Degree
and many-and-small
few-and-large
52
in the size
of
little
sample units, the use
measures restricts opportunities
with other studies.
for
of
Table 3. — Examples
of designs
quadrats
for subsampling
are not included
Substrate Class
individual
because
sites for percent cover of lichen species.
Studies
of problems with comparison with other studies.
Lower outer branches
Large,
low branches
Lower-
to mid-canopy
of open-grown
of open-grown
branches
in
Sample unit number, size, and arrangement
Specific case
Branches
using frequency
trees
5 5-yr and 5 10-yr branch segments
(Denison and Carpenter 1973)
trees
on oak and ash
100 equidistant points at 3 cm intervals on tape
spiraled around branch (Denison and Sillett 1990)
24 1-m branch lengths cut from midpoint of branches
closest to points at 4 m intervals along 3 parallel
transects (Lesica et al. 1991)
in conifer forests
Rock
High lichen cover,
Ground
Mostly
sparse
flora on forest floor
100 10x30 cm quadrats/site, regular intervals on
parallel transects (McCune and Antos 1982); 45 20x50
cm quadrats at regular intervals on parallel transects
(Lesica et al. 1991)
Trunks
Mostly
sparse
flora near cities or large point sources
1 m cylindrats, 10 trees per tree species per site;
certain tree species used (McCune 1988; Muir and
stone walls and natural outcrops
Point sampling on 10 cm grid, variable number of
points per rock face (1.1m), three rock faces per
station (John 1989)
McCune
1988); 2 m cylindrat,
10 trees
per site
(Johnson 1979).
Both rich and sparse
floras in a regional study
British acidification study: 100 evenly spaced grid
points in 18x27 cm quadrat with wire grid; subjective
quadrat placement, permanently marked, certain tree
spp. (Quercus and Fraxinus preferred); variable height
and aspect; variable number of quadrats/site (1-20)
(Looney and James 1990; Wolseley and James 1990).
Fairly rich flora in deciduous
forest
25x25 cm quadrat on NE aspect at 1.4m on 10 trees
per host species per site (Will-Wolf 1980).
Cover Classes
have been devised. Some of the most common
Using cover classes rather than attempting
logical
to
estimate cover to the nearest one percent tends to
speed sampling
similarity
are not based on cover
classes have been shown to be effective
scales is probably
of these
surrogates
for direct biomass measurement (Hermy 1988,
McCune 1 990), and are good detectors of community
are summarized
consistent differences
of these
for broad-scale
5. Use
the large plots.
A Broad-Scale Survey Method
between observers (Sykes et al.
A
gradient strategy of regional
The most useful and commonly used cover classes
arc narrow at the extremes and broad in the middle.
in Chapter
These approximate
large plot without
which is generally desirable for
sampling
is given
we present an extreme case of the
sampling strategy using a single
5. Here
"few-and-large"
an arcsine-squareroot
presented below
subsampling.
is an example
The method
of
this strategy. This
method would be most useful in a broad-scale
data. The cover classes can thus be
homogeneity
in table
studies using single large plots as sample units,
1983).
analyzed directly,
have also been used. Several
most appropriate
without subsampling
changes through time (Mitchell et al. 1988). A
disadvantage of cover classes is the potential for
converting
and/or
high degree of
among the systems.
similar to unclassed data, provided that the classes
(Sneath and Sokal 1973). Cover
are not too broad
proportion
the
Other Abundance Classes
Methods for field scoring lichen abundance that
than is realistically
they yield statistical results that are
transformation,
4. Note
and data entry. Cover classes do not
pretend to more accuracy
achievable;
in table
are listed
survey
of lichen communities. The advantages and
disadvantages of this general approach are
improving normality and
of variances among groups, without
to percentages. Many cover class schemes
summarized
in table 2. The method was tested in
1991-1992 (McCune and Lesica 1992) as part of the
53
Table
4. — Cutoff points for cover classes. Question marks for cutoff points represent classes that are not exactly
defined as percentages, but substitute another criterion, such as number of individuals.
Cutoffs in parentheses are additional cutoffs points used by some authors.
square-root
Notes/References
Cutoff points, %
Name
Arcsine
0, 1,5, 25, 50, 75, 95, 99
Designed
to approximate
percent cover (McCune
squareroot transformation of
1988; Muir and McCune 1987, 1988).
an arcsin
Braun-Blanquet
0, ?, ?, 5, 25, 50, 75
Uses two categories of low cover not exactly defined as percents.
1965, Mueller-Dombois
Commonly used in Europe (Braun-Blanquet
and Ellenberg 1974).
Daubenmire
0, (1), 5, 25, 50, 75, 95
Widely used in western U.S. in habitat-typing
Service and many other studies (Daubenmire
Carpenter 1973).
Domin
0, ?, 1, 5, 10, 25, 33, 50, 75
One category of low cover not exactly defined as percent
and Ellenberg 1974).
(Krajina1933; Mueller-Dombois
Hult-Sernander
Modified
(.02, .05, .10, .19, .39, .78); 1.56,
3.13, 6.25, 12.5, 25, 50, 75...
Based
Table 5. — Abundance
System
Henderson
System
References
Ministry of Forests
(1988) in Geiser et al. (1989)
Canadian
individuals
1 = Rare (3 individuals)
2 = Uncommon (4-10 individuals)
3 = Common (10 individuals but less than half of the boles or branches
have that species present)
4 = Abundant (more than half of boles or branches have the subject
species present)
McCune (1992)
1 = very rare ( 1% cover, approximately)
2 = uncommon (2-10%)
3 = common (11-25%)
4 = abundant (26-50%)
Henderson
et al. (1989)
(50%)
of all
pilot studies for Forest Health Monitoring (FHM)
consists
under the
plants, excluding
program
1976).
but with a few gaps
and dense
5 = very abundant
EMAP
(Oksanen
not based on cover estimates.
1 = Rare, a single individual
2 = a few sporadically occurring individuals
3 = a single patch or clump of a species
4 = several sporadically occurring individuals
5 = a few patches or clumps
6 = several well-spaced patches or clumps
7 = continuous uniform occurrence of well-spaced
8 ■ continuous
9 = continuous
FHM pilot
halving of the quadrat
Description of classes
Name
Canadian
classes
on successive
efforts by U.S. Forest
1959, Denison and
(general information:
macrolichens
occurring
on woody
the 0.5 m basal portions
of trees
of
Messer et al. 1991). The FHM method was designed
to be "user-friendly" and usable by a field crew with
minimal training in lichenology. Key components of
the method are that the field crew distinguishes
accurately assign species names to the lichens
among species and assigns coarse abundance classes
done later by a specialist),
in the field, collects vouchers of each species in each
distinctions
plot, and defers the naming
of
and shrubs. Second, an estimate
each species is made in the field.
member responsible
of
abundance
Note that the crew
for this task need not be able
to
(that is
but must be able to make
among species.
species to a specialist.
The first objective of the field sampling is to make
collection of voucher specimens for identification
by a specialist. The collection represents the species
diversity of macrolichens on woody plants in the plot
as fully as possible. The population being sampled
Recommended
Field Procedures (about 1-1.5
hr for 1 person):
a
•
The area to be sampled (henceforth the
"Lichen plot") is a circular area with 36.6
m (120 ft) radius.
54
•
Take a reconnaissance
lichen plot, locating
woody
walk through
o
the
plants, and collecting
samples and estimating
•
voucher
3.
abundances as you
Collect lichen species with fruticose and
foliose growth forms (i.e. macrolichens).
Inspect all trees and shrubs 0.5 m tall
4.
visible symptoms, and
tree cores) for lichens.
to inspect the full diversity of
How to Handle Uncertainties
The field crew will frequently have uncertainties
substrates present: trunks and branches,
hardwoods
and conifers,
large shrubs.
•
about the classification
to inspect the full range of
Be careful
microhabitats
present: shaded and exposed,
topographic
uniform slope, so long
within the lichen plot).
•
Record
positions
(for
in a draw, on an otherwise
example,
as the draw occurs
relative abundances within the
lichen plot. Relative
1.
3.
4.
2.
When the growth form is in doubt, assume
it is a macrolichen.
a
lichen.
assume that two different
mail the bags to
Bags should be packed closely,
but without excessive crushing, in sturdy cardboard
(4-10 individuals in area)
specialist.
boxes.
of
Common (10 individuals in area but less
than half of the boles and branches have
field crew to make as many distinctions
that species
as possible.
The purpose
present)
by excluding
Abundant (more than half of boles or
Collect
have the subject
species
present.
in the field
can later adjust the data
specimens that are not macrolichens
forms that were considered
separately by the field crew but are actually
conspecific.
Upon receiving the samples,
open the boxes immediately
a
the specialist
should
and check for damp
lichens. If some are damp, they should be thoroughly
air dried. The specialist should then identify the
contents of each bag by species. In the case
collections,
see the special
instructions
of
mixed
in McCune
(1992). The specialist should then record species
name(s) on the bag and on the data sheet (appendix
5
of this chapter) and return the contents
original
Place in #2 brown paper bag labelled with
appropriate codes:
species number (sequentially
these rules is to encourage the
The specialist
and by combining
palm-size (about 5 cm in
diameter) sample of fruticose and foliose
growth forms. This includes all species
that are three-dimensional or flat and
lobed. Even minute fruticose and lobate
forms should be included. In many cases,
a much smaller sample will be obtained
because of the scarcity of the species.
o
2.
the lichen
Sample Procurement
1.
When in doubt, assume it is
Once the samples are collected,
Rare (3 individuals in area)
branches
The
forms are different species.
Code:
2. Uncommon
an organism.
1.
3. When in doubt,
abundance for each
species is estimated using the following
Abundance
of
following rules for the field crew are designed to put
the responsibility
for classification on the specialist,
not the field crew.
upper branches and lower branches, and
trees in particular
Specimens must
decay. If specimens were wet when
collected, the individual bags should be
spread out and dried inside or in the sun
as soon as possible.
samples (the same trees used for
foliar nutrient analyses, branch and foliage
fallen branches,
Store bags in a dry place.
be thoroughly air dried to avoid fungal
destructive
Be careful
Place all voucher bags from one plot in a
larger brown paper bag. Record plot ID
code and date. Staple or tape top of large
bag closed.
within the lichen plot for lichens. Also
inspect branches collected for the
•
(revise as collection
proceeds)
go.
•
relative abundance
lichen epiphytes on
bag. The bags and lichens
for future reference.
as col
lected)
o plot ID code
55
to the
should be stored
Data Analysis - Quantitative Community Data
1.
Analysis of quantitative data on lichen
communities relative to air pollution can have several
components:
of
differences
associated
of
isolation
differences
summary statistics,
descriptive
description
major gradients and community
differences
with those gradients, and
related to air pollution from
related to other sources. Each
topics is considered
of
2.
these
below.
Descriptive Summary Statistics
1.
Site by species table — A straightforward
summary of community data is a table of
species abundances for each site with row
and column totals or averages. Species
abundances may be site-level averages if
sites were
2.
subsampled.
-- The site by species
with columns
or rows representing species richness,
such as average number of species per
sample unit and total number of species
Diversity statistics
table can be supplemented
recorded
3.
at the site.
Components of variance -- A helpful but
infrequently used method for describing
scales of variation within a hierarchical
sampling scheme is to partition the total
variation of a response variable like
species richness into variation contributed
at different scales. The calculations can
be done using nested ANOVA, available
in most standard statistical packages. The
method can be used to describe the
relative importance of different sources of
variation or to formally test the
hypothesis of no difference at a given
level, based on the F-ratio.
relating
the
most common
Preliminary Data Adjustments
Perform
an "Index
of
Atmospheric
Ordinate
associated
IAP
ordination
with
smaller number (typically 1-3) of composite variables
of the variance in the original n x
p matrix.
Relate sample scores on each dimension
technique for
Purity"
ordination
is to
correlation,
on a
the weighting
it cannot be recommended
the
are elucidated
and graphical
by
overlays.
of ordination techniques
of technique becomes important
as the heterogeneity of the data set increases.
Methods were reviewed by Will-Wolf (1988).
scheme. Although in some cases this method can be
used effectively,
These relationships
regression,
of
and site
There is a wide menu
as a weighted-average
with many variants on
space to species abundances
characteristics.
(IAP).
the species occurring
is calculated
the Samples in Species Space
representing much
Many different indexes have come under this rubric,
site. The
and
Reduce the n sample unit by p species matrix to a
but most are site scores based on the average
diversity
any needed data standardizations
transformations.
Interpret Ordination
data to air quality
lichen community
calculate
The method does not attempt to separate
community variation caused by
differences in air pollution from other
sources of community variation affecting
species diversity. In an area thought to
have pristine air quality, differences in
community composition can be
attributable to variation in environmental
factors and disturbance history. In areas
with a gradient in air quality, community
differences due to variation in air quality
must be separated from differences due to
other factors. This is usually done by
extracting the dominant compositional
trends in the data using an ordination
technique, then determining if any of
those trends are related to air quality.
The basic steps in this procedure are described in
Ludwig and Reynolds (1988), Greig-Smith (1983),
and Gauch (1982). Briefly, the steps are:
Analysis of Spatial Gradients
Historically,
IAP
values will be found in
of the presence or
absence of differences in air quality.
Therefore, the name "Index of
Atmospheric Purity" is potentially
misleading.
Differences in
any data set, regardless
available.
The choice
Currently
the most popular
plant community
in
data
ordination
are Detrended
techniques
for
Correspondence
Analysis (DCA), Nonmetric Multidimensional Scaling
(NMS), and Bray-Curtis ordination (BC). NMS is a
general for reasons listed below:
robust, relatively
56
assumption
free method that is
suitable for community
generally
Sorenson group of distance
to air quality and components
data, using the
of variation. There
measures. Computer
are only
related to other sources
tools for
a few basic
programs to conduct these analyses are available.
separating potential air pollution effects from other
Chapter 5, Sensitive
factors affecting
lichen communities,
many variations
on those tools. Six main tools are
Species, also explores gradient
analysis and ordination.
along with examples
listed below,
Community Changes Through Time
Two data sets from the same community taken
two different times will always differ to some
degree. Three components
of
1.
at
observed differences
through time are: (1) actual changes, (2) visual
estimation error, and (3) spatial variability. Actual
changes may be due to changes in
community
among species. Visual
or interactions
environment
error is unavoidable,
estimation
but can be measured
close in time, and hopefully
by repeated sampling
will be small. Spatial variability in communities is an
intrinsic part of even the most seemingly uniform
communities. With repeated random sampling, some
of the observed differences through time will be
derived from spatial variability. This source of
variation can be virtually eliminated by using
2.
of
differences
of
of
variance
the factors
powerful than
Repeated measures
within-subjects
are
(ANOVA)
3.
using
not appropriate,
and
ANOVA.
a repeated-measure
ANOVA
designs)
(also known as
focuses on the differences
through time for each sample unit.
Assuming that changes in permanent plots
found that exceed differences
are
expected from
of the changes
of observed
differences must be made on the basis of many
sources of information, including data or observations
on changes in air quality or other environmental
measurement error alone, the direction
should be interpreted.
variables,
successional
Interpretations
changes, disturbance
and known ecological characteristics
showing
of
history,
the species
4.
Lichen community monitoring provides correlative
results rather than proof of causal relationships,
replicated experimental
is part of a controlled,
manipulation
of air quality.
Because that is rarely the case, we must rely on
inferences based on the literature on lichen responses
a careful
These inferences
must be linked to
attempt to separate community
whether spatial or temporal,
differences,
into a component
results based on known relative
sensitivities of species to air pollutants.
Depending on the study area, much or
virtually nothing will be known about the
particular species. Existing compilations
of species sensitivities (Wetmore 1983,
Ryan and Rhoades 1991) contain only a
small fraction of North American species,
of data from
reflecting the preponderance
Europe and, to a lesser extent, eastern
North America. Nevertheless, in most
Interpret
of
the species
will
before to allow one to
infer whether or not a compositional
trend is related to air quality.
Isolating Air Pollution Effects
unless the monitoring
trends
to air quality by using other tools
study areas, enough
to air
the changes (i.e., sensitivities
the main compositional
have been studied
pollutants).
to air pollutants.
Separate
listed here.
through time should be analyzed. T-tests
time as one
are less
Remove factors unrelated to air quality by
specifying them as covariates in an
ANOVA or by removing them first in a
multiple regression. For example, if
species diversity on trunks is related to
trunk diameter and air quality, the portion
related to trunk diameter can be
controlled first, using that variable as a
covariate or preceding the entry of air
quality variables into a regression (e.g.
McCune 1988).
related
permanent plots changes the way
or standard analysis
of their use.
(axes) in the data using an ordination
technique (see also Chapter 5), then
determine which, if any, of the axes are
permanent plots.
Use
but there are
related
57
Correlation with direct, instrumental air
quality data at sampling points (e.g.
McCune 1988) is ideal, but this approach
has seldom been used because so few of
these stations exist in most areas. Direct
instrumental monitoring is so expensive
that, outside of urban/suburban areas, an
investigator is lucky to have even one or
two direct monitoring stations as
reference points.
used in subsequent analyses should be marked in
should be studied. The frequency
species in sample space. The resulting
of
Urech.
data
Press. Pay particular
describing
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the
by the University
of
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of
Plant sociology:
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Pollution
Crock,
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Determining
Gough,
1991.
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for easy reference. UTMs or
references for each site must be
Photographs
be provided;
1965.
(Transl. by G. D. Fuller and H.
Conard) Hafner, London. 439 pp.
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techniques used
plots in later years. Make sure that the map includes
provided.
in
epiphytic lichens in the vicinity of Fox Creek and
in an appendix. All study area
locations should be described in great detail and
marked on topographic maps or copies of the
suitable landmarks
Advances
51:401-406.
processing
should be included
relevant sections thereby enabling
and M.
of deposition
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Multivariate
eight different air pollutants to lichen data in
Braun-Blanquet,
meet the standards established
Manual of Style, published
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Aerobiology
by biological journals. Most follow The Chicago
Chicago
1988.
small town in Switzerland.
S.
reports should
K., R. Herzig, L. Leibendorfer
Ammann,
See Muir and McCune (1988)
REPORT REQUIREMENTS
All
it.
LITERATURE CITED
related to the air
a
species sensitivity.
for an example.
to
of
species ranks
gradient can be used as a measure or ranking
quality
funds were available
methods (e.g. BC,
a separate ordination
or scores en axes that are strongly
and
and rationales useful
P.
one can construct
if
management action
implement
J.
NMS)
options are
assessment of the
in preparing the environmental
method DCA.
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managers with evidence
Provide
to the air quality
are provided
by the ordination
management
correlated with evidence presented in the report.
J.
Species ordinations
gradient.
automatically
If
available.
they should be stated concisely
provided,
species in sample space can be used to
species sensitivities
future
Management Recommendations
If future work is being recommended, the cost and
timing should be included here with a succinct
rationale. This information will help managers to
program budgets or seek funds if they are not
Where there is a clear gradient in air quality,
of
of
should also be discussed.
monitoring
immediately
assess relative
why it was not
Future Work
This section should include recommendations on
additional data that needs to be collected or sites that
Determining Species Sensitivities
ordination
of
some way with a brief explanation
used.
time.
through
computer files, should also be
in the appendix. Any data collected but not
included
can also be used to make
inferences about changes
etc. Original data, on field
data sheets and in
Temporal patterns relative to known
changes in air quality can also be used.
When air quality changes rapidly, as with
the startup or closure of a point source,
strong inferences can be made with a
Before/After /Treatment/Control design.
See Stewart-Oaten et al. (1986) and
Eberhardt and Thomas (1991) for an
in-depth discussion of this class of
designs and potential problems with
pseudoreplication. Repeated Measures
ANOVA
should list the statistical
techniques
package used, version,
Water,
Air
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of ecology 71:437-450.
Journal
Tabachnik,
power generating plant.
air quality improvement.
Use of visual cover assessments as
1983.
of air
Showman, R. E. 1981. Lichen recolonization
Ecology 67:929-940.
in time?
quantitative estimators of some British woodland
Ohio and Parkersburg, West Virginia. Ohio Journal
of Science 73:357-363.
quality around a coal-fired
and K.R. Parker.
Sykes, J. M., A. D. Horrill, and M. D. Mountford.
flora of the Ohio River Valley between Gallipolis,
as indicators
Ministry of
impact assessment:
Environmental
1986.
Showman, R. E. 1973. The foliose and fruticose lichen
Showman, R. E. 1975. Lichens
Quantifying
Forests. (ms.)
Forest, Springerville.
National
1991.
arboreal lichens for habitat management: a review of
Brook Lakes Watershed, Sierra
on lichens 1986-90.
(Natural History)
1990.
The effects of
London:
British
43 pp. + unnumbered
Chapter 4 Appendix
1
TRANSECT DATA SHEET
TRANSECT
DESCRIPTION
Direction of reading (e.g., Left to Right, or Top to Bottom)
device set points (positions
Measuring
A
Transect
set points:
INTERCEPT
of nails or stakes along the tape or meter stick, holding it in place):
B
End
Beginning
Transect
length
LENGTHS (in centimeters)
SPECIES
Total
SPECIES
Total
TOTALS
% COVER
total cover
transect
x 100%
=
FREQUENCY
length
FOOTNOTES:
COMMENTS:
62
Chapter 4 Appendix
2
LICHEN PLOT DATA SHEET
SITE NAME
DATE
PLOT ID (alphanumeric
TIME
SURVEYOR(S)
code)
(include affiliation)
LICHEN SPECIES (for each species, give scientific name or acronym, collection number of voucher, color and other
features
used in field identification)
SUBSTRATE (record kind or name, size or diameter, condition, surface features, and modifying factors, e.g., rain
tracks, or adjacent branches)
PLOT TYPE (e.g., dotiometer transect, photographic quadrat, etc.)
POSITION OF PLOT (height above ground, inclination, slope and aspect, etc.)
SITE DESCRIPTION
(beyond
information on general site description form)
DIRECTION OF PLOT (compass
PHOTOGRAPHS
COMMENTS
bearings,
landmarks, trail references,
markers)
(roll and frame numbers; subject; direction; where taken from)
(include diagram of plot and measurements
of selected thalli)
63
Chapter 4 Appendix
Recommendation
methods.
However,
- The
following dichotomous
3
can be used as a decision tree for recommended sampling
in this tree, since many situations
key
it is important to retain more flexibility than can be indicated
will dictate an approach different than that recommended here. The letter following each entry in the key
corresponds to the rows in Appendix 4, "Synopsis of Recommended Methods."
1a.
1b.
Rapid, broad-scale survey method needed, to be done by field
crew with minimal lichen training
Whole plot/voucher method
-A
Intensive, more precise method needed, to be done by field crew with good lichen identification
2a.
Sampling on trees or shrubs
3a.
Trunks
4a.
4b.
Communities
-
Objective is to monitor well-developed
examples of target communities
British fixed-point method — D
Branches
6a.
Open-grown
7a.
trees and shrubs
Objectives
allow small datable branches
branchlets
E
-
Equal-aged
7b.
6b.
Objectives require study of larger branches
Branch centers
F
Forest trees
8a.
Tall forests without easy branch access
-
-
8b.
2b.
B
Communities mostly dense
5a.
Objectives require random or otherwise
non-subjective sampling
Trunk quadrant pairs — C
5b.
3b.
mostly sparse
Trunk Cylindrats
G
Twig pickup
or Litter pickup
H
Good access to lower- or mid-crowns
Branch centers
F
-
Sampling on ground or rock
9a.
Mostly sparse cover
Simple quadrat — I
9b.
Dense cover
or Fixed points, ground
Simple quadrat
to lichen
- (application
biomonitoring
64
-
I
not known)
skills.
Chapter 4 Appendix 4
Synopsis of recommended methods.
given particular study objectives.
B. Trunk
cylindrat
Plot, 36.6
Circular
m (120
Single
radius
ft)
plot/
voucher
Arrangment of SU's
Sample Unit (SU)
Name
A. Whole
Use the preceding key as a guide to which methods might be appropriate
References
McCune 1992
plot, random location in
stand
homogeneous
t2, 1-m half
Randomized
cylindrats per tree,
0.5-1 .5m high on
tree. Average both
values
Typical # of SU's per site
McCune 1988, Muir
within specific tree
criteria (e.g. particular
tree species or size range)
10 per target tree species
or 24 per mixed species
or nearest trees to
10 per target tree species
of 24 per mixed species
stand
Lesica
10-20 grids per site, 100
Wolseley and
James 1990;
selection
& McCune
1988
stand
from half
quadrat
0.2
x
C. Trunk
2,
cylindrats
0.5 m
Randomized
flexible quadrats on
opposite sides.
the
Average
pre-determined points along
parallel transects
et al. 1991
2
2
pairs
quadrats
Points
x
Grid point
18
27 cm quadrat
in
D. British fixed
point
in fixed-location
trunk, subjective
grid on
grid placement
points per grid
Looney
1990
E. Equal-aged
branchlets
Variable-Length
branch segment
known
of
age
Randomized,
one branchlet per
tree or (original method)
subjective selection of twigs with
20 or 10 (original
method)
and James
Denison
&
Carpenter
1973
most lichen cover, all branches
from same tree
centers
G. Twig Pickup
m long branch
segment centered
on midpoint of
Branches
1 m long and 2-8 m
above ground, choosing branch
nearest to vertical line from
branch
pre-determined
transects
Fallen twigs or
10 cm
branches
Branch with highest lichen cover
4 x 1 m box centered and
equidistant on eight 13 m radii
1
F. Branch
long
Litter picked
circular
10-20 depending
canopy
along parallel
standardize sampling to
when violent storms are
0.2
0.5 m or
m quadrat;
smaller quadrat
more dense
Equal or randomized
parallel transects
1
x
quadrat
Randomized
season
et al. 1991
Geiser 1991
transects;
rare
x
Simple
sorted
Lesica
points on parallel
weighted
1
I.
2m radius,
plot,
&
Lichen
up
in
H. Litter pickup
10 per target tree species
or 24 per mixed-species
stand
intervals on
for
communities
65
45
on
McCune 1991
uniformity
Lesica
et al. 1991
Chapter 4 Appendix
5
Lichen Identification Data Sheet
Date:
Plot No.
Lichen Specialist
# = Bag number
A = Abundance rating
Data from collecting bags
Data to Enter
Species Name
Sp. code
A
A
#
A
n
#
A
#
A
Comments
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Abundance
codes:
4 = abundant
Species codes:
1 = rare (< 3 individuals
seenV
(more than half of the branches
Refer to master
list for 4-character
2 = occasional
M-10
individuals
seen have this species).
lichen species
code names.
66
seen).
3 = common
(>10 individuals
seen).
Identification of Sensitive
Species
Jayne Belnap, Lorene Sigal, William Moir, and
Sharon Eversman
We describe procedures for determining the responses and sensitivities
of lichen species to air pollutants. Pollutant chemicals in their primary
forms and as derivatives such as ozone or acid rain can cause acute and
chronic
injury to lichens and other cryptogamic plants. Fumigation
studies are designed to reveal plant responses to air pollutants under
controlled
in enclosed exposure systems, generally through
conditions
changes in physiological
Gradient studies are designed to show
locations along an air pollution gradient. They
processes.
responses at predetermined
measure visible damage and are done around existing
or proposed
Both types of study have limitations, but are useful for
detecting air pollution effects and tracking changes in existing or proposed
pollution sources, as well as for understanding natural succession and
usually
sources.
pollutant
variability.
OVERVIEW
Air
pollutants
are formed
can cause both acute and chronic
effects on lichens
and other organisms
mosses and cyanobacteria.
such as
liverworts,
Atmospheric
deposition
and concentrations
to vegetation,
injuries
such as
Angeles
are
well documented
(Shriner
of
et al.
these
of biological concern in
they are emitted.
a result
of
pollutants
(O3),
pollutants
of
compounds.
Ozone and
referred to as
agricultural
Semi-volatile
and persistent organics
are
processes.
High concentrations of toxic organics and pesticides
have been found in fog and in lichen tissues
(Glotfelty 1987; T. Moser, personal communication).
form as
are created as
Studies have demonstrated
nitrate (PAN), and acid rain are
pollutants.
organics,
transported and deposited via atmospheric
are
reactions involving primary
transport in the atmosphere. Ozone
secondary
1983), the central
trace metals, and metalloids) are of
concern as well (Moser et al. 1992), although there is
limited information on the effects of these
1990; Nash 1988,
same chemical
spots due to
pesticides,
chemical
during
peroxyacetyl
examples
the
Secondary
and Nash
"air toxics" (e.g., industrial
pollutants
1972; Ferry et al. 1973; Gilbert 1973).
Primary pollutants such as SO2 and fluorides
basin (Sigal
documented cases (Shriner 1986).
Other types of pollutants, commonly
Historically, sulfur
(SO2) and fluorides have received the most
study, and acute effects to lichens and other
around point sources
and necrotic
ozone exposure occur in the Los
(Shriner et al. 1990). Acute foliar injury to
vegetation from acid rain is rare, with few
dioxide
vegetation
involving
Sierra Nevada, and in the eastern United States
generating plants, gas purification plants, metal
smelters, aluminum production plants, cement plants,
plants, and pulp mills.
such as bleaching
acute and chronic
necrosis, morbidity, and mortality, are usually found
only around point sources. Examples of large point
sources include fossil-fuel fired steam electric
chemical
reactions
NO2 are oxidized to sulfuric acid (H2SO4) and nitric
acid (HNO3) during atmospheric transport. Visible
of air pollutants great
enough to cause acute injuries
in photochemical
nitrogen oxides (NOx) and hydrocarbons mostly
emitted by vehicles.
Acid rain is formed as SO2 and
affect lichens (Nash
PAN
67
that various
1972, 1975).
trace metals
A
of
number
other pollutants,
known
correlation
to be present
but for which there is little or no information,
are
of
various
analysis,
significant agreement among the
scales is shown and this strongly
supports the
less interest for one or more of the following
inference that lichens actually
reasons:
(Nash 1988). For other air pollutants, data are
currently insufficient to establish precise sensitivity
1)
pollutants are widely distributed, but
seldom
cause damage
because
scales.
ambient
hydrogen fluoride,
The information
cause damage
For areas in
affect resources of a Class I area.
which there are no data, the information provides
baseline or control data to which subsequent data on
For areas
effects of new pollution can be compared.
are rare (e.g., hydrogen
al.
et.
1990).
of
allow comparisons
air
multiple
of
symptoms
plant stress such as chlorosis
(loss
At
chlorophyll, yellowing) and reduced growth.
level, sensitive
community
from the population.
Lists of species known
under certain conditions
been published
pollution.
of
(Nash and Greis
the
to be
The most widely
combination,
used
studies did not address proof
of
However,
stirred tank reactors (CSTRs), open-top
Pollution studies
without
these
Air
cause/effect
fumigations,
have been used to develop quantitative
relationships
between concentrations
plant responses, thereby establishing
cause/effect
relationships.
The most commonly
independently
laboratory
established
physiological
to be:
acidic
rain
measured responses are
processes sensitive
nitrogenase activity,
electrolyte
Lichen
to fumigation appear
K+ efflux/total,
leakage, photosynthesis,
and respiration
pigment status, in order of sensitivity
(Fields 1988).
with fumigation studies
depends on such factors as
There are still problems
fumigation
For SO2, scales of sensitivity
information.
such as Zonal
Study (ZAPS) and simulated
selected physiological processes.
of air pollutants
and various
Controlled
done in the field
are occasionally
chambers, using techniques
Pollutant
chambers,
cuvette chambers.
exposures.
More recently, fumigation studies involving some
variation of a closed chamber with air movement
through the chamber, or chamberless field
with gradient studies give
and in
in more or
conditions
branch chambers, and miniature
relationships.
coupled
under controlled
less enclosed exposure systems such as continuously
studies.
Historically, gradient studies usually involved
observations of visible injury, species richness, or
species abundance correlated with pollutant exposure,
when exposure data were available.
STUDIES
Fumigation studies are designed to show
measurable responses to air pollutants, singly
of lichen
methods are gradient studies and fumigation
studies
and
changes in air quality.
FUMIGATION
have
1991).
the responses and sensitivities
species to air pollutants.
air pollution sources, continued
of
significance
In this chapter we review procedures for
determining
of
measurements can show the magnitude
species may disappear
sensitive
to pollutants
patterns, the natural
species, and the effects of existing air
Following permitting, construction, and
of
variability
operation
may
to
over time that contribute
successional
understanding
with urban development, such as
small stationary sources and large numbers of mobile
sources. Chronic injury is less likely to involve
death of tissues, but is likely to involve nonspecific
associated
pollutants
data, the information
where there are historic
Chronic injury develops after long-term or repeated
exposure to sub-acute concentrations
from a proposed facility may unacceptably
emissions
the high concentrations necessary
to cause damage
to
of Significant
(PSD) permits, to show that estimated
Deterioration
sulfide, hydrogen chloride) (Shriner
to fumigation
such as ozone,
and zinc (Nash 1988).
collected from gradient and
for Prevention
applications
to vegetation
affect
studies can be used in relation
fumigation
3) pollutants are widely distributed, but
seldom
sensitive
species that are demonstrably
experiments with air pollutants
2) pollutants are not widely distributed,
occurring only as accidental releases (e.g.,
chlorine, ammonia); and/or
measured
concentrations
pollutant
in the field are high enough to adversely
exposures have mean ratios and mean
concentrations that are too low to affect
lichens (e.g., ethylene, carbon monoxide);
because
However,
do respond to SO2
because sensitivity
the best
have been
based on controlled
concentration
and duration
environmental
conditions,
of
exposure,
and status
of
the thallus
during exposure (Fields 1988). In addition to the
very important influence of thallus hydration level, it
fumigation results and on field
(Nash 1988). By non-parametric
observations
68
is known that different
of
portions
vary in
a thallus
•
Karenlampi
environment
ambient conditions.
resembling
In addition, the accurate determination
their physiological response
(Nash et al. 1980,
1970; Moser and Nash 1978; Moser et
Long-term
laboratory
of
are not currently
problems
However,
during an experiment.
considered
in maintaining
viability in growth chambers.
procedures which should be followed before and
studies to
fumigation
injury
determine chronic
feasible because
controlled
specimen
short-term
exposure-response
lichen material does not generally
regulatory
do well for even
of
and their
specimens
agencies in developing
build for individual
and Nash 1984, Pearson and Benson
be possible
Pearson
determining
physiology
(e.g., photosynthesis,
studies have used only single pollutants
respiration)
and ultrastructure.
the factorial
experimental
mixtures because of limitations
number of exposure chambers available.
and synergistic
antagonistic,
two or more pollutants
are known
in
Controlled
the
However,
•
responses to
for vascular plants
activity,
•
exposure capability;
concentration
conditions
requirements
throughout
non-reactive
•
precise control
pollutant
injected
into
conditions
light, relative humidity)
in these systems. (Shriner et al.
Mini-Cuvettes
throughout
An additional form of exposure chamber used
for a number of years is a cuvette which
encloses a branch or a portion of a branch in a
chamber
successfully
surfaces;
of
of
system,
1990).
and between chambers;
•
most widely accepted system is the
(e.g., temperature,
is not reproduced
chamber and between chambers;
environment
to
within the
chambers. The primary concern is how well the
results reflect plant response under field conditions,
since the dynamic nature of field environmental
for plant exposure systems, whether they be
controlled environment, greenhouse, or field, are
listed below in order of their importance (Heagle and
Philbeck 1984):
pollutant
the
the system and uniformity
there is no perfect
Essential
controls
stirred tank reactor (CSTR)
which ensures rapid mixing of pollutants
have become more sophisticated
However,
boxes, bell jars or even plastic
by adding environmental
Currently,
and Philbeck 1984). Over
time, systems for studying the effects of single or
system.
available
chambers to
continuously
(Heagle
pollutants
commercially
exposure chambers.
exposure systems range
parameters and pollutant
(Lange et al. 1984).
all-purpose exposure
can be attained in
bags) within a single controlled-environment
chamber; or
from plastic bags (Anderson 1976) to elaborate
chamber systems with automatic control of
uniform
on lichen
nitrogenase
by putting smaller exposure chambers
(plexiglass
Gaseous and particulate
•
by converting
pollutant
•
Exposure Systems
uniform
exposure conditions
controlled-environment
less complex.
•
of pollutants
three ways:
(Shriner et. al. 1 990), and there is no reason to think
that lichen responses to multiple pollutants are any
multiple
the effects
Controlled Environment Chambers
designs necessary to study the responses of plants to
concentrations
at existing
Most laboratories are not
equipped to perform
environmental
Chamber
to
it may
studies are most useful for
Although air pollution is a complex phenomenon
involving multiple contaminants, most fumigation
additive,
However,
to conduct experiments
facilities.
under given
too expensive
lichen studies.
1970).
of conditions.
dose-response
of air quality standards
replicates, but are usually
multiple
physiological condition throughout the study (Link
1977,
for other
(Drummond and Pearson 1979). The chamber
systems described below have the advantage of
short times in artificial chambers, attention must be
paid to the treatment
important
are
curves for the establishment
Since
relationships.
and
techniques
reporting
researchers working in related areas and for
studies (less than a month) are valuable
for explaining the mechanisms of plant response and
establishing
Properly designed and
experimental
comprehensive
fumigation
pollutant
pollutant
and between chambers requires acceptable calibration
al. 1983).
sets
of
parameters within
and environmental
concentrations
mature tree canopy (Legge
et al. 1977, Lange
et al.
1992). This approach permits
the measurement of mechanistic plant tissue
concentrations;
1984,
and
69
Amundson
et al.
in situ in a
responses, such as photosynthesis,
canopy, eliminating
of
the effects
many
controlled
small chambers
of
the concerns
variations
In
(Eversman
exposure chambers.
it is possible
of SO2 at ground level and vertical
in SO2 concentrations within plots
or no pooling
regarding
1979).
to introduce pollutant
gases in sub-parts per million as part of the inflow
Acid Rain Simulation
using the flow-through air exchange principle.
However,
of pollution
this type
rainfall
Contaminated
exposure has not yet
been used with lichens.
from watering cans to back-pack
irrigation
Open-Top Field Chambers
ambient environment
as closely
while
within
as possible
allowing control of pollutant concentration
chambers.
Appropriate
non-chambered
available
currently
predictive
control
The open-top
developing
the
and ambient chambers as well as
pollutant-free
effects.
of all of the major
of dose on a programmable basis
(Shriner 1979). Rain simulators have been used in
combination with a ZAPS-type fumigation system
and are being used with open-top chambers to study
the combined effects of acid rain and gaseous
chamber systems include
using open-top
purposes
natural precipitation
technique for
•
useful for
•
•
ZAPS
Systems, referring
pollutants
in
the
of
Air Pollution
delivering
between
of
controlled
ions to a deionized
water
stream.
air
field for relatively long-term testing
with no control over environmental
two systems described
dispensed
GRADIENT ANALYSIS
The
conditions.
only SO2.
Moser et al. (1980) used a small system at
Anaktuvuk Pass in Alaska to fumigate caribou forage
lichens Cladina stellaris, C. rangiferina, and Cetraria
cucullata.
Anhydrous SO2 was dispensed through
measurable attributes of affected plants vary along
causative environmental gradients. This assumption
is illustrated
in figure
paired holes along an aluminum
relationships
among the importance
concentrations
method assumes that
The gradient analysis
Sulfate
pipe.
were measured at intervals upwind
downwind from
1,
which shows unimodal
values of three
lichens and one moss along a moisture gradient
and
the pipe.
(Flock 1978).
One large system (ZAPS I) was operated by the
US Environmental Protection Agency from
Species are best represented at their environmental
optima, and will not be found at limits outside their
1975-1979 in Custer National Forest in southeast
tolerances.
Montana
to predict possible
effects
of
grassland ecosystem.
A
operated from 1976-1979.
of
such as grazing intensity,
locations
set at
sulfation
levels with elevation
of
disturbance
factors
fire regimes, or trampling.
of
lichens
contaminants.
above ground level. In
basin.
general, less SO2 was recorded at the lowest level,
10 cm, than at the upper two levels, indicating little
sources like the Los Angeles
The most commonly
variables
are
Studies are
or projected sources of
The sources can be point sources like
power plants or diffuse
in
at predetermined
along an air pollution gradient.
usually done around existing
per plot, adjacent to transplanted lichens.
slight differences
include climate and
Gradient analysis can be designed to show
measurable responses
SO2
10, 50, and 100 cm above the soil surface at five
locations
lichens
substrates, as well as gradients
a
system (ZAPS II)
Both ZAPS I and ZAPS
In 1978, sulfation plates were
The plates demonstrated
In addition to air pollution, other
gradients influencing
duplicate
had four plots, each with different
concentrations.
from
emissions
new coal-fired power plants on components
II
of
droplet size range (0.1 - 3.2 mm);
chemical composition, through metering
concentrations
stands for Zonal
to means
controllable
(intensity
pumps which supply carefully
ZAPS Systems
The acronym
rate
characteristics
including:
0.5 and 2.7 cm/hr);
et al. 1990).
(Shriner
rainfall
systems can
and chemical
reproduce the physical
chambers are the best
relationships
The more sophisticated
pollutants.
plots to estimate any chamber
experimental
functional
sprayers and
to very elaborate systems
sprinklers,
components
designs for
experimental
the
designed to permit variation
chambers used in the field duplicate the
Open-top
in
has been simulated
using a variety of approaches,
field and laboratory
measured response
visible injuries such
thallus deformation,
as bleaching
changes in community
and
structure
such as species richness or cover, and physiological
70
Buellia punctata
u
Cladonia gracilis
a>
o
Niwot Ridge, Colorado
Cladonia pyxidata
c
•Q-
.*
Polytrichum junip,
O
Q-
E
V)
o
0)
•■»■•
D_
C/J
\r
s-..
Dry
Moisture Axis
Figure 1. — Importance of 4 cryptogams along an indexed
processes such as photosynthesis,
integrity
leakage), pigment quantity,
degradation,
(electrolyte
and
Gradient
from
dry to wet (from
Flock 1978).
of
microclimate,
substrates,
or extremes of weather variability.
The theory of gradient analysis is well developed
trampling,
is appropriate
analysis
of air
loadings
moisture gradient
more direct effects
fluorescence.
effects
Wet
This last problem can be especially vexing, since
pollution impacts on lichens may be minor relative to
nitrogenase activity,
element uptake, membrane
n
pollutants
on lichens,
for studies of
the
(Ter Braak 1987). Measurements are taken across a
variety of environments, and differences among the
since pollutant
often vary with distance from point sources.
It is not necessary for other environmental
measures are assumed to be controlled
factors to
by
vary continuously
in the real world in order to use
environmental
gradient analysis.
There are often sharp
can be analyzed in several ways, which are
discontinuities
Gradient
in spatial or temporal dimensions.
analysis
is also useful for analysis
regional effects derived from non-point
this case, the regional
summarized
of
sources. In
Reynolds
airshed is assumed to be large
Direct
of lichens
of which
demonstration
(1988) for mathematical details.
gradient analysis is used where lichen
in gradient studies has some
the researcher
should be aware
of
measurements, or long-term
exposures
to pollutants
for example, by air quality maps. Measured
attributes of lichens or other cryptogamic plants are
plotted along an elevation, soil pH, or pollution
as given,
of
for the particular
•
from the treatise by Ter Braak and
Readers are referred to Ludwig and
of occurrence, or
or physiological symptoms are
described as a function of measured environmental
variables. Each sample can be associated with its
elevation, soil pH, on-site air chemistry
species;
determination
Data
morphological
(Johnson 1976). These include:
• difficulties associated with identification
•
between the sites.
abundances, probabilities
exposure levels.
The use
here
Prentice (1988).
enough that there is wide latitude in pollutant
limitations
differences
the best indicator
species
study; and
that the observed distribution
loading axis.
The relationship
patterns reflect pollution stress and not
variable and the environmental
other biotic and/or abiotic factors.
determined by regression.
71
between the lichen
quantity can be
Gradient
composition
community
environmental
calibration
of
as a measure
If
problem.
of
the variation
Morphological analyses detect change to the form
or structure of an organism. A disadvantage to
morphological analyses is that not all pollution injury
This is indirect gradient analysis, an
problem according to Ter Braak and
Prentice.
Ludwig and Reynolds
as "a set
of
techniques
(1988) define
in which sample
will
axes such that their relative positions
the axes and to each other provide
information
Indirect
about their ecological
gradient analysis
of known sensitivities
pollution effects
effects
of
Gradient
A
of
variety
1977,
various
Hill
multivariate
analyses, are discussed
An advantage to
analyses is that they can be done with
These types of
and funds.
analyses can also be done with still or video
cameras. Comparisons can be made before and after
treatments, throughout
and
time at the same sites, and/or
along a distance gradient from a known pollution
source.
data set, and
multivariate
Microscopic
by Ter Braak and Prentice
(1988) and Ludwig and Reynolds
size, and
the species chosen.
limited instrumentation
by
visual assessment of lichen
such as coloration,
of
macroscopic
software
derived from the various
the unaided
appearance
variables.
methods to use on a particular
interpretations
by
such as
DECORANA,
1979,
This is
characteristics
when subtle
ORDIFLEX,
CANOCO (Gauch
Ter Braak 1988). Which of the
packages are available,
PC_ORD,
Macroscopic
are present.
is widely employed
analysis
ecologists.
microscopically.
is most useful when lichens
other environmental
Morphological changes can be analyzed
without optical aids, or
macroscopically,
maximum
likely to be overwhelmed
are
to
similarities".
to air pollutants
The method may be less effective
produce a response that can be distinguished
visibly.
units are arranged in relation to one or more
coordinate
each
Morphology
in species
composition.
ordination
of
from the
to compute abstract axes that
species assemblage
ordination
and/or advantages
limitations
Specific
and physiological
characteristics.
and population
technique are noted here.
variables are
not measured, one can use information
account for much
characteristics
are
to this as a
environmental
measured in air pollution studies
include individual morphological
values when the regressions
Ter Braak and Prentice refer
known.
Characteristics
is also applied by using
analysis
Microscopic analyses allow for observation of
injuries that may or may not be otherwise
visible.
Two types of microscopes are used in
light
analysis of injury assessment in lichens:
(1988).
cellular
ANALYTICAL TECHNIQUES
microscopes
Many analytical
examining
are available
techniques
the effects
of air
pollutants
for
effects
on lichens.
Which techniques are appropriate for a given study
will depend on the type and levels of pollutants
involved, the lichen species used, the nature of the
work, for example)
The analytical
listed below.
techniques
in fresh mounts
that
protocols
to both
and levels
of
of
the percentage
lichens exposed to SO2, showing
of
to estimate the algae's
plasmolysis
health.
green and round; plasmolyzed
increased
and other recommendations,
the status
cells are
Healthy
cells are yellowed and
Kauppi (1980) demonstrated
shrunken.
usefulness
including quality assurance, quality control, levels of
injury
the
SO2 on algal
with increasing exposure time.
Observations of fresh mounts use color and shape of
chloroplasts, which comprise most of the algal cell,
field and laboratory work, unless otherwise indicated.
Using any of these techniques will require additional
literature research to determine their suitability for
needed, specific
the proposed work, instrumentation
procedural
of
Will-Wolf
stress, especially
significantly
used most often are
applicable
pollutant
microscopes.
in estimating
Eversman (1978) and
(1980)
estimated the percentage of plasmolyzed algal cells
and the resources available.
These are generally
has been useful
cells.
studies as opposed to fumigation
research (gradient
of
electron
and transmission
Light microscopy
of fluorescence
of algal cells.
microscopy
the
in determining
An alternative method to determine dead and
plasmolyzed
exposure.
sections
of
(Holopainen
72
algal cells uses fixed and embedded
lichens stained with toluidine
and Kauppi 1989).
Empty
blue
cells are
Pigments
interpreted as dead; cells with shrunken contents are
Chlorophyll and its degradation
Again, increased
was observed with SO2 fumigation.
considered
plasmolyzed.
plasmolysis
Transmission electron
microscopy is a helpful tool to visualize or verify
cellular effects when physiological effects have been
observed. Many characteristics of chloroplasts and
mitochondria change in intermediate stages of injury;
severe injury results in near collapse of all
recognizable cell structure (Eversman and Sigal,
1984, 1987; Holopainen and Kauppi 1989). Some
specific changes recorded are: deformation of
thylakoids and pyrenoglobuli of the chloroplast,
swelling followed by degeneration of mitochondria,
unusual accumulation of starch granules and lipid
of
recognizable
1985,
1987,
1976,
mimic ordinary
1981,
and
(Beekley
and Harper
Belnap
1990, Garty et. al.
Henriksson and Pearson 1981, Kardish et. al.
LeBlanc and Rao 1973, Nash 1973, Nash
Ronen and Galun 1984, Eversman 1980, Moser
et al. 1980). Researchers have traditionally
effects on chlorophyll as total chlorophyll
concentrations,
concentrations,
in chlorophyll
chlorophyll a:b ratios, or the ratio of
and degradation products.
can
chlorophyll
The more commonly
of these pigments in dimethyl
sulfoxide
Extracts
(DMSO).
chlorophyll
at optical
are then
levels of
scanned to determine
density 435 nm and
it is important to have fresh material and to control
conditions under which samples are stored before
fixation so that moisture and light conditions of the
degradation products at optical
control and treated samples have been identical.
techniques to do so include the use of red light
all pollutants seem to cause the same
ultrastructural changes in algal cells so one cannot
It does seem, however,
distinguish among pollutants.
during sample preparation,
such as O3 and
standardization
and in
Nitrogenase Activity
here.
Most
All
the more commonly
require some level
are sensitive
instrumentation.
(Belnap
to seasonal or geographical
and care must be taken
nitrogen usable by vascular
contributors
plants.
of
1978,
in which they occur (West
Evans and Ehlringer unpublished
data, Denison 1973). Atmospheric pollutants have
been shown to affect nitrogenase activity levels
Bergman
which should be taken into account when
It is important to recognize that lack of
sampling.
homogeneity within a lichen thallus has been
documented for various characteristics (e.g.,
photosynthesis, nitrogenase activity, chlorophyll
experimental
of
are
converting
nitrogen to the ecosystems
variation,
concentrations),
nitrogen,
and Skujins
to
used ones are listed
of
phycobionts
These lichens may be important
determine physiological changes resulting from air
pollution;
Lichens with cyanobacterial
capable of "fixing" atmospheric
it into a form
Physiological analyses detect alteration to the
normal functioning of an organism or any of its
available
and
(1960).
Physiology
There are many techniques
cold storage, and
time between extractions
For further details on these techniques, see
Brown (1980), Brown and Hooker (1977) and Vernon
less time than SO2 and acid treatments (Eversman
and Sigal 1987, unpublished data).
parts.
of
analysis.
PAN
cause cellular damage at lower concentrations
density 415 nm
(Ronen and Galun 1984). It is important to reduce
chlorophyll degradation during sample analysis;
Unfortunately,
that the oxidant pollutants
used
method is extraction
spectrophotometrically
seasonal stress or normal senescence,
reported
percent reduction
chlorophyll to its degradation products.
There are several means of quantifying
organelles
stress
affected by pollutants
significantly
Hoffman
and membrane structures.
Since cellular changes due to pollutant
are the
Studies have shown chlorophyll levels to be
Some cells are too small to accurately observe
with light microscopy.
bodies, and disappearance
products
pigments most often used to assess pollution injury.
1990, Denison et. al. 1977, Hallbom and
1979,
and Pearson
Hallgren and Huss 1975, Henriksson
1981, Kallio and Varheenman
1974,
Sigal and Johnston 1986).
Measurement of nitrogen fixation is difficult and
Sheridan
1979,
costly. Measuring
nitrogenase
fairly simple and cheap.
if
In
nitrogenase enzyme, acetylene
material does not include the entire
ethylene.
Consequently,
activity,
however,
the presence
of
is converted
nitrogenase
activity
is
the
to
levels
in the amount of ethylene that is
produced. Levels of ethylene and acetylene can be
Incubation in an
measured on a gas chromatograph.
acetylene atmosphere can be done in the field or the
laboratory, using gas-tight containers with an
thallus (Karenlampi 1970, Nash et al. 1980, Moser
and Nash 1978, Moser et al. 1983).
are reflected
73
10% acetylene under
Membrane
light and temperature in the
Exposure
atmosphere of approximately
of
standard conditions
in the field (see above
lab, or ambient conditions
rates
lichens exposed to increasing
levels has
employ
of
methods
et al. 1983). Membrane leakage can be estimated by
measuring the conductivity of de-ionized water before
measuring
and after immersion
1978, 1979, 1980;
Fields and St. Clair 1984a, 1984b).
common
of
repeatedly in the literature
et al. 1971, Eversman
using
potassium
(Beckerson and Hofstra 1980, Hart et al. 1988,
Pearson 1985, Pearson and Rodgers 1982, Belnap
and Harper 1990, Fields and St. Clair 1984b, Moser
Respiration (Gas Exchange)
A significant decrease in respiration
(Baddeley
of
efflux, total electrolyte
leakage, and scanning electron microscope work
can be used to transport samples (Rychert
been demonstrated
has been demonstrated
measurements
and Skujins 1974).
pollutant
This
lichens.
and trace metals has
membrane integrity in
been shown to compromise
In the field, portable chambers can be
references).
placed over the experimental material, and
vacutainers
Integrity
to sulfur dioxide
the lichen.
Instrumentation
rates
Elemental Analysis
This is covered in Chapter
an oxygen electrode to measure O2
or an infrared gas analyzer to measure
CO2 evolution in the absence of light.
absorption
7.
Population Characteristics
Photosynthesis
These are species characteristics
rates have been determined using
Photosynthetic
which can
chambers and gaseous 14C02 (Fields and St. Clair
1984a, Hallgren and Huss 1975, Ross and Nash
measured in the field and require little or no
1983, Sigal and Johnston 1986, Lechowicz 1982),
quadrats as samples.
in liquid
CO2 (Fields and St. Clair
or
measured
as
the rate of CO2 absorption or
1984a)
O2 evolution (Beekley and Hoffman 1981).
characteristics,
instrumentation.
immersion
Determining
Karenlampi
availability of an infrared gas analyzer (IRGA), while
with
gross photosynthesis
CO2 labeling
techniques requires a liquid scintillation counter.
Chlorophyll fluorescence is a set of response
variables associated with the light reactions of
photosynthesis, particularly those associated with
photosystem II. It is a convenient, sensitive, rapid,
systems most commonly
the fluorescence
light.
behavior
analyzed
rise to maximum
This induction
of
rates, mortality,
cover, and biomass
can be
These measurements can
1959; Armstrong
1971) or by photography.
whether done with a traditional
Photography,
camera
Details are
provided in Chapter 4. Table 1 provides an example
of studies done with different pollutants and the
effects
of
those pollutants
on chosen lichen species.
of
The experimental
levels at saturating
altered after
is measured
or a modified
design of studies addressing
sensitivity
of
be similar
to any research design.
lichen species to air pollutants
method should be determined
problem,
gas
the questions
to be answered,
lichens have found decreased photosynthetic
initial hypotheses or working questions.
Determining
of
rates
which
the
the
anticipated use of the data, site characteristics
available
resources.
the
should
The choice of
by the nature
exchange system (T. Nash, personal communication).
All methods which measure photosynthesis in
with exposure to pollutants.
1991;
1970,
EXPERIMENTAL DESIGN
is the kinetics
Fluorescence
using a chlorophyll fluorometer,
made using
photosynthetic
curve is markedly
exposure to air pollutants.
be
such as reproductive
or with video, can be computer-analyzed.
way to assess photosynthetic
The fluorescent
growth
be done by eye (Daubenmire
systems depends on the
are
Variables
measured within quadrats.
estimating
efficiency.
Measurements
presence/absence,
net gas exchange in either flow-through
or closed chamber
and non-invasive
is
meter, which is both cheap
and portable.
The most
respiration
of
limited to a conductivity
First, decide upon a set
and
of
broad
Then do a
literature search to determine the lichen species in
of
methods to use, (instrumentation required, replication
and portability for field work) is discussed in Link et
the targeted area, the sensitivity
al. (1984).
Related research might include work with the same
the air pollutants
in question,
species in the same locality
pollutants,
techniques.
74
these species to
and other related work.
or with
the same
and/or using the same analytical
Table 1.
— Selected authors
and papers describing effects of various pollutants on lichens.
Source: Eversman and Sigal (1985).
Effect(s) observed
Pollutant
Sulfur dioxide
Baddeley et al., 1973
Puckett et al., 1973, 1977
Tomassini et al. 1977
Richardson & Nieboer, 1983
Fields & St. Clair, 1984
Holopainen & Karenlampi, 1984
Respiration decreased
Membrane permeability increased
Permeability increased (ions lost)
Review: physiology, ecology
Permeability increased, photosynthesis decreased
Ultrastructural changes
Ozone
Brown & Smirnoff, 1978
Rosentreter & Ahmadjian, 1977
Nash & Sigal, 1979
Ross & Nash, 1983
Sigal & Nash, 1983
Eversman & Sigal, unpub. ms.
No effect on photosynthesis
No effect on chlorophyll content
Photosynthesis decreased
Photosynthesis decreased
limited, morphological changes
Distribution
Ultrastructural changes, photosynthesis decreased
PAN (peroxyacetyl
nitrate)
Sigal & Taylor, 1979
Eversman & Sigal, 1984
Photosynthesis decreased
Ultrastructural
changes
Nash, 1976
Loss of chlorophyll pigment
NOx
Fluoride
Nash, 1971
LeBlanc, Comeau, Rao, 1971
LeBlanc, Rao, Comeau, 1972
Roberts & Thompson, 1980
Takala, Kauranen, Olkkonen, 1978
Loss of chlorophyll pigment
Plasmolysis, color change, loss of chlorophyll pigment
Species die, disappear
Discoloration, morphological changes; species disappear
Morphological changes; species disappear
Acid precipitation
Denison et al., 1977
Robitaille, LeBlanc, Rao, 1977
Lechowicz, 1982
Lechowicz, 1984
Sigal & Johnston, 1986
Sigal & Johnston, 1986
Eversman & Sigal, unpub. ms.
Nitrogen fixation decreased
Loss of species
Photosynthesis decreased
Growth decreased
Photosynthesis decrease
Photosynthesis, nitrogen fixation
Ultrastructural changes
75
decrease
Available resources should be identified.
Resources
the range
variables across the widest range
access to study areas, collaboration
transportation,
and any other factor that may influence
possibilities,
This can be done by narrowing
of soil, topographic, grazing, or other
as much as possible.
time, equipment,
people, dollars,
include
your capability to carry out the intended research.
From the collected information, develop a set of
be obtained
refined hypotheses.
feasible.
These hypotheses are a
that are feasibly
answerable
will
species used, analytical
chosen, and resource availability.
to most study designs
possible,
techniques
Elements
at places where
levels occur, with a
pollutant
indirectly
common
can be made by using known sensitive or
This is the calibration
species.
referred to above.
of
Samples
near the high, low, and middle portions
depend on the
questions being asked, the types of pollutants
and
If
resources. Make a list
of likely candidates from an inventory of available
species and species known to be sensitive to the
Select species from the list
pollutants in question.
based on the questions being asked and available
resources. Another option is to run preliminary
analyses on species that are selected for reasons
other than known sensitivity, such as abundance or
ease of collection, to ascertain whether they are
to the pollutants
the
Heyligers
sampling
sampling
variation
anthropogenic
effects
well
as
The response
of
influences
communities
of pollution (Jackson
secondary
of
with known sensitive or accumulator
of
of
the pollutant.
species are
sites can be established
cryptobiotic
across a
in different
Particular
communities
in Chapter 4.
of
the
sample communities,
since these
may have a large effect on cryptobiotic species
Sciences 1987, Hawksworth
(Northrup Environmental
influences.
and Rose 1976, McCune et al. 1987).
the subtle
1990).
Any
Sample Size
of samples
of sites. Sample size
depends on the variability of the characteristics being
measured and the degree of confidence with which
with the influence of other variables.
To minimize effects of confounding factors, we
recommend a sample design which incorporates
known pollution levels.
If the pollution source is local and fixed and the
pollution "shadow" is known, sampling should be
spatially explicit with respect to the source.
Considerable environmental variation will still
remain, which again may overwhelm possible
influences
as
gradsect sampling
sensitive or accumulator
sampling
microclimate
be confounded
secondary
If
effect of pollution upon lichens is likely to
a sample design
Therefore,
airsheds as discussed
natural or
and Gough
a region.
attention should be given to the substrate and
techniques
may overwhelm
in
across a region as with floristic variation
wide range
to more direct
environmental
can be limited to one or several regionwide
unknown,
as to anthropogenic
lichens
1989). This is a
across the most important
affected by pollution.
present.
(Chapter 4). Lichens respond to natural gradients
moisture, temperature, light, nutrients, and biotic
competition
we
(Austin and
method of efficient
The concern is not so much with total floristic
can vary from survey data to
sampling
pollution source is regional or non-local,
features that control species distributions
Gradient Sampling
Gradient
the
suggest using gradsect sampling
species.
more elaborate vegetation
of
gradient.
species present, and available
sensitive
analyses
(Chapter 7). These samples should also be spread
across the gradient, or if this is not practical, then
Species Selection
suitably
problem
the accumulator
species can be harvested for chemical
will
no
are
then an attempt to measure pollution
accumulator
include:
The species selected for study
If
few near the middle (to test for linearity).
direct measurements of pollutant loadings
Sample design
can be
only a few direct measurements
the highest and lowest
given available
on geography,
vary greatly depending
accessibility,
If
at as many
obtained, they should be concentrated
resources.
Sample design is the next stage.
pollutant
of pollution loadings should
of the sample sites as
Direct measurements
distillation of the initial broad hypotheses into
questions
of
loadings.
Therefore,
Sample size includes both the number
per site and the number
comparisons
between samples can be made.
Data
from a pilot study are useful in determining
variability
and then computing
to detect possible
use
that limits other sources of variation
76
adequate sample sizes
changes (testing null hypotheses).
Repeatability
fail because sites
are
lost or different
analytical
techniques are used that cannot be calibrated
Repeatability influences the timing of sampling
Factors to consider are the
and site selection.
to
previous techniques.
of the organism, variability in pollutant
phenology
levels, physiological condition of the organism,
of
aspect and substrate
slope,
LITERATURE CITED
sites, distance from
(in topical categories)
factors such as large bodies of water,
influencing
roads, etc., and any other factors that may affect
study results.
Overview
Confounding variables that cannot be
need to be identified.
eliminated
Ferry, B. W., M. S. Baddeley and D. L. Hawksworth.
1973. Air Pollution and Lichens. Athlone Press,
Plot Protocol
Plot protocol determines
London.
the way in
or future samples are taken.
which current
Gilbert, L. L. 1973. The effect of airborne fluorides. In
Air
Permanent plots in
unwanted influences
indirect trampling
on materials.
Glotfelty,
1987.
Athlone
Press, London.
D. E., J. N. Seiber and L. A. Liljedahl.
Pesticides in fog. Nature 325: 602-605.
Moser, T. J., J. R. Barker, and D. T. Tingey.
For example,
may affect soil lichens.
B.W. Ferry, M.S.
ed.
pp. 176-191.
may need to be restricted at sites to prevent
activities
and Lichens,
Pollution
Baddeley, D.L. Hawksworth,
which no destructive sampling takes place must be
distinguished from temporary plots from which
material is collected.
Designation of sampling areas
within temporary plots may be desirable so that the
location of disturbance is known. Certain types of
Anthropogenic
contaminants:
1992.
Atmospheric
transport,
deposition, and potential effects on terrestrial
Methods
for marking and mapping plots must be identified.
Current options for establishing coordinates for study
vegetation In Science of Global Change: The Impact
of Human Activities on the Environment, American
sites include topographic
Chemical
maps and global positioning
Series 483. Dunnette, D. A. and R. J.
O'Brien (eds).
Collection Techniques and Material Treatment
Nash
Collection techniques and handling, transportation,
and storage of collected
use
of
the material.
different
If
analytical
In addition, requirements for
. 1975.
work is to be done, pre-treatment
protocol and the physiological condition of the
fumigation
. 1988.
procedures
techniques
are employed,
using statistical calibration
design studies with this in mind.
current study will be available
monitoring
Consider
in the future.
manual with instructions
personnel.
and
Air
Quality.
J. Cramer,
30: 201-216.
1991.
Lichens
as
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Environmental Chemistry, Volume IV, Part C.
Hutzinger, O. (ed). Springer- Verlag, New York.
III, T.
1980. Sensitivity of
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effects of air pollutants on Mediterranean and
Nash
H. and L. L. Sigal.
lichens to air polluUon
find ways
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desired
Riverside,
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a monitoring
studies with field
fumigation
Bryophytes
45: 183-198.
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should be kept as simple as
new techniques
data
Correlating
Nash III, T. H. and C. Gries.
It is desirable to design studies so that results can
with past and current studies. To
facilitate valid comparisons with future studies, field
to compare
315-324.
Berlin-Stuttgart.
be compared
If
the Blue
near a zinc factory.
Influence of effluents from a zinc factory
Bibl. Lichenol.
Comparison with Past, Current, and Future
Techniques
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material before and after experimental
p. 498.
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III, T.
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Monitoring systems commonly
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surface. Water, Air, and Soil Pollution 11: 191-200.
In
W. W. Heck, S.
Usnea, and Umbilicaria.
Air, and Soil Pollution 27: 315-322.
Interaction between simulated rain and barren rock
13.
Linzon (eds.). Air Pollution
Pseudoparmelia,
G., A. Stuanes and K. Bjor. 1979.
C. and D. W. Larson.
deposition
Effects on Vegetation.
S. N.
76:
simulated acidic rain on one species each
Acid Rain
Bailey,
lichens from high
for the Assessment of
Sigal, L. L. and J. W. Johnston,
Abrahamsen,
53-66.
43:
of Botany
Journal
of Methodology
Control Association,
survey relative to oxidant
64:
American
V. Krupa and
on conifers in Southern California
an ecological
of
Shriner, D. S. 1979. Atmospheric
nitrate fumigations.
Sigal, L. L. and T.H. Nash III. 1983. Lichen
communities
epiphytic
a copper smelter.
elevation cloud forests as indicators of atmospheric
564-575.
82:
of
et Lichenologique
and chemical studies
studies on the gross photosynthetic
of
to the paucity
Scott, M.G. and T.C. Hutchinsoa 1989. Morphological
80: 600-605.
Sigal, L. L. and O. C. Taylor.
lichen
Water, Air, and Soil
G., F. LeBlanc and D.N. Rao. 1977. Acid
Revue Bryologique
and the
simulated acid
the caribou
18: 421-430.
cryptogams in the vicinity
Effect of
ozone on the lichen Cladonia arbuscula
in
Brodo.
(Opiz.)
rain: a factor contributing
1-59.
Rosentreter, R. and V. Ahmadjian.
Bryologist
stellaris
of
The effects
1982.
precipitation on photosynthesis
Botany 23: 71-77.
Ross, L. J. 1982. Lichens
M. J.
Lechowicz,
lichens. Environmental
(L.) Hoffm. Environmental
26:
and Experimental
Botany
59-64.
R., B. Caldwell, B. Bormann, L. Eldred, C.
Pollutant Mixtures
Swanberg and S. Andersoa 1977. The effects of
acid rain on nitrogen fixation in western Washington
forests. Water, Air, and Soil Pollution
coniferous
Nakagawa, Y. 1982. Effects of sulfur dioxide and
8.
nitrogen dioxide alone or in combination
21-34.
chlorophyll
Fritz-Sheridan,
R. P.
1985.
and P. polydactyla.
Hill, D. J.
1971.
tinctorum. Taiki Osen Gakkaishi
Impact of simulated acid
rains on nitrogenase activity in Peltigera
Sigal, L. L. and J. W. Johnston,
17: 27-31.
Lichenologist
Experimental
aphthosa
study of the effect
New Phytologist
Lechowicz, M. J.
of
of
stellaris.
Bulletin of
America,
65: 144.
of
the caribou
the
1986.
of
Effects
and
photosynthesis in the lichen Lobaria pulmonaria
(L.) Hoffm. Environmental
831-836.
The effect
1984.
rain on the growth
70:
Jr.
17: 370-376.
acidic rain and ozone on nitrogen fixation
sulphite on lichens with reference to atmospheric
pollution.
on
content and enzyme activity in Parmelia
26:
simulated acid
lichen Cladina
Ecological Society of
88
59-64.
and Experimental
Botany
Active Monitoring
Lorentz C. Pearson
Active monitoring
of air
controlled
transplanting,
quality using lichens may involve
fumigation,
or culturing of lichens.
Transplanting
from areas where they occur in nature to
involves
moving
localities
where they are needed to monitor air pollution or other
of
characteristics
organisms
the environment.
Controlled
fumigation
consists of
creating a point source of pollution that can be manipulated
of
to test the
on lichens that occur in the area.
Culturing
of lichens in areas where they
occur in nature and/or developing methods of growing lichens in gardens,
farms, or laboratory growth chambers under controlled conditions.
Active monitoring experimental treatments must be carefully planned,
effects
specific
or enhancing
is encouraging
replicated,
pollutants
and randomized.
the growth
The material to be transplanted must be
healthy specimens of the biomonitor
lichen material to
is impossible
which they
Methods
species.
the substrate must not be harmful
or impractical
to move them
are attached. We recommend
of attaching the
to the lichens when it
with pieces of the substrate to
where possible,
that,
lichens be
to the same kind of substrate from which they were taken.
We also recommend using a combination of evaluation methods to
transplanted
estimate the degree of environmental
degradation caused by the suspected
pollution source.
INTRODUCTION
measurements
of lichen
evaluate the quality
Because lichens are especially
sensitive to air
area.
pollution and can accumulate and concentrate toxic
substances from the environment in their thalli, they
are recognized
by scientists
as valuable
where they naturally
These measurements
include
are used to
in the target
of
observations
in the target area
(Brodo 1961, 1966; Holopainen 1983, 1984; Skye
1958), various physiological responses of the lichens
biomonitors
(Pearson and Skye 1965, Pearson 1980, Holopainen
1984), and accumulation of pollutants in the lichen
tissues (Lawrey
and Rudolph 1975, Nieboer et al.
of air quality with lichens
1978,
lichens from a source area
of air quality with
in gardens
or farms or encouraging more rapid or abundant
At present, this
growth by using culture techniques.
Rope and Pearson
Theoretically,
occur to a target area where
1990).
active monitoring
lichens may also involve
pollution is suspected (Brodo 1966, Hawksworth and
Rose
characteristics
the environment
how well the lichens survive
of atmospheric quality. Biomonitoring can be either
passive or active. Passive biomonitoring is covered
in other chapters in this book (See chapters 2, 4, and
5). Active monitoring
involves transplanting
of
1976, Skye 1979).
Active monitoring may also involve observing
lichens in areas where they naturally occur following
the installation of a factory or other source of
pollution (Skye 1958, Will-Wolf 1980, Ryan 1990,
Kandler 1987); controlled fumigation is sometimes
employed as the source of pollution (Pearson and
Rodgers 1982). In either case, periodic
is largely an unexplored
ADVANTAGES
growing
lichens
field.
OF ACTIVE MONITORING
The chief advantages of using transplants
monitor air quality
89
are:
to
•
mercury, copper, cadmium,
being able to plan and set up orthogonal
experiments
locations
with sampling plots
at
all
and to observe the nature
where it is desired to monitor air
A
quality;
zinc, ozone, carbon
hydrogen fluoride, or peroxyacyl
monoxide,
number of problems
of
the
nitrates)
resultant injury.
may arise when conducting
•
having enough plant material at each
studies involving active monitoring.
anticipated
•
location to run all of the desired tests;
having uniform material collected from
common site at all of the sampling
potential problems
a
•
insufficient supply of a test species of
•
variation
•
transplant shock;
lichen;
being able to choose the best experimental
design for the particular
and
location
material;
•
ascertaining
how rapidly
increased levels
of
pollution cause injury in lichens.
The ability to study the effects of different
conditions
environmental
by planning
whether
seasonality
•
choice
•
given source
of
the extent
of
pollution.
The information
can be supplemented
any injury
with
by fumigation
of
transplant;
species;
between pollution level and
the relationship
lack
of
content
of
the lichens;
baseline data or historical
(herbarium) information
abundance in the area.
is indeed causing the observed injury, and to evaluate
more accurately
of
the elemental
where the
pollution source
a suspected
due to ecotype differences;
•
•
samples are placed enables the researcher to judge
more accurately
Some
with transplant studies
to consider
include:
locations;
•
These must be
and steps taken to avoid them.
on native lichen
caused by a
gained
METHODS
a known
For example, Pearson and Rodgers (1982)
placed four sulfur dioxide burners in areas of dense
low-lying forest, dense upland forest, marsh with
pollutant.
scattered trees, and open dry grassland.
levels
of sulfur dioxide
Transplant
Planned
1)
were produced by burning
sulfur on days when the wind speed and direction
Upwind sites provided the
were appropriate.
unpolluted
An
controls.
often overlooked
advantage
of
transplant
has a wide variety of
designs from which to choose. Often a
studies is that the investigator
experimental
Another
of
advantage
active monitoring
to occur, regardless of whether
or not they are adapted to the area, for
the purpose of accumulating pollutants in
lichen tissues and observing physiological
This latter method is sometimes
changes.
called the "moss bag" method.
expected
is the
In conducting transplant studies, we recommend:
• limiting the number of collection sources in
order to minimize variability;
to observe rapid responses to pollution or
other environmental
changes.
When abundant
material has been placed at each monitoring
subsamples
can be harvested weekly,
monthly, or at other intervals.
methods are sensitive
site,
•
biweekly,
enough that changes in injured
in a matter of weeks even
low levels of pollution, and in a matter of hours at
extremely
•
of
a variety
of
pollutants
moving
the lichen
comparable
•
with substrate attached
or transplanting to
substrate;
if physiological
studies are to be made,
using species well within their normal range
When fumigation is used to supply a point source
of pollution, it becomes possible to evaluate the
effects
species that are easily collected,
whenever possible,
at
high levels.
(sulfur dioxide,
choosing
transplanted and observed;
Some evaluation
lichens can be observed
Transfer of healthy lichens from an area
where they occur naturally to an area for
which they are adapted, to study possible
changes in physiological processes or
accumulation of heavy metals or other
elements as a result of exposure to the
pollution (Brodo 1966), and
2) Transfer of large amounts of healthy
material to an area where pollutants are
randomized block design is best, but lattice designs,
Latin and Greek squares, and other designs often not
available to other types of field research can be
employed in transplant studies. This is especially
See
true in experiments involving fumigation.
Federer (1955) for descriptions of experimental
designs and advantages and disadvantages of each.
ability
methods may be classified in two
categories:
of adaptation both
target area;
lead,
90
at the
source and at the
•
using historical
better than expected, but did not survive
records, such as herbaria or
check lists where available,
natural occurrence
to study the
of species in the target
area (Thomson 1970);
• where possible, using species
of
known
whenever
possible,
collecting
areas that are essentially
transplant
the need
for such observation
modified
if
each specimen at the time of transplant, and
frequency
taking an initial sample for elemental
•
every month for the first year.
photographed
site in every way but air quality;
• measuring thallus dimensions, photographing
should be measured and
lichen transplants
Ideally,
to the
the data show
that a longer or shorter
of
We also recommend that the number
treatments be chosen carefully and
experimental
if
adequate replication
glue like araldite (Hawksworth
and Rose 1976) or liquid nails; if it is
4 replications
desired to tie lichens to shrubs, trees, or
minimum
at
We recommend
planned.
least 5 or 6 experimental
non-toxic
Later,
should be assessed and
is more appropriate.
baseline;
it is necessary to use glue, using a strong,
the lichens.
and Experimental Design
Measurement
lichens from
identical
the second
glues held well, but
substances that injured
produced volatile
sensitivity;
•
Other silicone-type
spring thaw.
treatments and at least 3 or
in most transplant experiments
of 25-30 experimental
units.
with
in mind that
posts, avoid string that rots when exposed
material must be transplanted,
to sun or water.
some transplants may die, become detatched from
their substrate, fertilized by birds or other animals,
do not collect
Generally,
transplants
minimize
desert lichens for
This will
in microclimate
in the high mountains.
effects of differences
Transplants
may be in the form
branches, twigs, or rocks or pieces
lichens attached.
vegetation
Specimens
preferred
of bark plugs,
of rock with
or placed on specially built poles.
that blend with the natural vegetation
or rubbed loose or
eaten by animals.
Based on estimates of potential
losses, the amount
of
transplant
material needed can
be estimated.
To minimize
following:
These may be attached to existing
to prevent vandalism,
of vandalism,
become victims
between the collection and transplant locations.
keeping
a
Adequate
•
minimize
transplant shock, we recommend
the
the time from collecting to
transplanting;
are
•
though they may be
difficult to relocate.
allow specimens to air dry before
transporting,
and do not transport in plastic
bags;
•
Adhesives
attach specimens at the elevation
For attaching clean specimens to objects in
conditions
the
target area, our first choice is always to take
substrate to which the specimen is attached.
to the elevation
were collected,
If
this
humidity
glue or string must be used.
at
which they
in mind that
keeping
and variability in humidity
We
recommend against cotton or paper string which rots,
and against wire or string containing metal. Fishline
generally decrease with elevation
seems to have no serious disadvantages,
the trunk
providing
above the
ground, and average daily temperatures
increase in the sequence N, E, W, S around
it
is tied on with a knot that will not slip.
of a
If
transplant studies are conducted
refrigerate
of evaluating accumulation
toxic fumes.
a rock;
the material cannot be transplanted
immediately,
In a study conducted by Chris Warner,
a student at Ricks College, "liquid nails," a silicone
Umbilicate lichens glued to
glue, met both criteria.
basalt rock with epoxy showed visible signs of injury
(change in color and much increased rate of
electrolyte leakage), whereas those glued with liquid
nails showed no sign of injury and remained attached
through two winters and summers. Elmer's glue held
or the sides of
if
Glue must meet two criteria: it must hold well,
even during rainy weather, and it must not give off
tree
•
it.
is not possible,
above
ground with the most similar microclimatic
for the purpose
of heavy metals or other
elemental pollutants, we recommend keeping them
away from metal objects during collection and
transplanting.
Factorial
designs involving both wind direction
and
distance from pollution sources can be used to set up
treatments (Federer 1955).
can help in choosing experimental
the experimental
Windroses
transects.
91
At least one of the experimental
variable
up the independent
those obtained from lower levels over
Ecologists make use of four kinds of
in conducting air pollution studies:
treatment.
controls
•
some specimens
transplanting
at the
•
occurred.
which in turn
(Pearson and Rodgers 1982);
reverse transplanting to ensure that observed
than the peroxisomes.
injury is due to the increased pollution and
not merely to a change in environment; and
Improved
transplanting
specimens
devices
monitoring
if
of
sensitive to injury from high levels
to sites
the mitochondria,
•
SO2 than are
more sensitive
are
did not seem
Photorespiration
to be affected by the high
of
levels
pollution.
now in which the
chambers are available
lichens may be exposed to air containing
measured amounts of known pollutants
close to mechanical
them, rather than to stagnant air.
any are available.
Clair (personal communication)
that exposure
Fumigation Studies
to high levels
exactly
flowing over
Studies by St.
and others indicate
of pollution
over a short
period of time results in injury comparable
Field fumigation studies are especially valuable in
monitoring air pollution (Pearson and Rodgers 1982)
because they make possible direct observation of how
specific physiological or morphological
changes
If an industry
correlate with specific pollutants.
exposure at low levels over a long period
claims
lichens.
the problems
the type
of injury
of injury
to identify
fumigation
caused by a specific
pollutant.
time.
Very little work has been done on culturing of
Under laboratory conditions, with whole
lichens in growth chambers of various types, Tobler
(1939), Kershaw and Millbank (1969),Pearson (1970),
Pearson and Benson (1977), Dibben (1971), and
observed, the claim can
be tested by using controlled
to
of
Culture Methods
that the substances they released are not the
ones causing
in the
in urban areas where high levels
of pollution
upwind from the point source of pollution
•
time; and
The study suggested that plastids are more
some specimens
transplanting
of
observed in the lichens
collected
shock are not being confounded
(Brodo 1966);
•
longer periods
the symptoms
flasks were similar to those in lichens
source
to ensure that pollution effects and
location
transplant
patterns similar to
should reveal metabolic
treatments making
should be a control
If
downwind from the suspected industry, there is
and Deslooves (1974) have had moderate
success by ensuring periods of drying between
strong evidence
periods of adding water.
it is the same type
varying
levels
peroxyacyl
Margot
at the site
that the industry is indeed the
polluter.
A related method consists
into laboratory
occurring
Ahmadjian
of transplanting lichens
dioxide,
from isolated mycobiont
hydrogen fluoride,
(Pearson and
See Chapter 5 for more methods.
of sulfur
dioxide
be possible
critics
in stagnant air in
environment.
has experimented
in the field
their
manipulation
(personal
with dispersal of
of
the
communication)
lichens on BLM
land in Idaho with some encouraging
results, but
much more research is needed.
some factors that were pointed out
It may be more rewarding,
however,
to encourage
because SO2 is very soluble in water
(Hodgman et al 1957), its levels in the
accelerated growth and reproduction
flasks were not as high as the critics
fertilizer elements may speed up the growth and
dispersal of species suitable for monitoring which are
already in place. We know of no published research
areas where they
assumed;
•
Rosentreter
of
dispersion
propagules through experimental
in the report:
•
and
lichens to study sites in large
amounts, and encouraging
flasks.
overlooked
to encourage lichen growth
by introducing
Rydzak (1969), Coppins (1973),
and others criticized the study because the levels of
SO2 were much higher than those occurring in
nature, even in highly urbanized areas. However, the
laboratory
cultures,
(personal
The results from these studies suggest that it should
Pearson and Skye (1965) reported altered patterns of
photosynthesis and respiration in lichens exposed to
high levels
and phycobiont
has obtained
communication)
growth in Peltigera species from individual isidia.
Lindstrom
nitrates, or other pollutants
Henriksson 1981).
Ahmadjian et al. (1980),
(1982), and Jacobs and
Ahmadjian (1971) have reconstituted whole lichens
chambers where they are exposed to
of sulfur
and Jacobs
it was hypothesized on the basis of studies
with other kinds of plants that high levels
of pollution over a short period of time
along this line.
92
already occur.
of lichens in
Adding water and/or
SPECIES
changes.
using a fine grain black and
Photography
of
white film can be used to measure growth
In Europe, numerous studies have made it possible
and development
lichen species into several categories
to classify
lobes
In some cases, it
of apothecia.
may be desirable to have photographs
taken monthly
ranging from the most tolerant to the most sensitive
over a period of a year or two.
(Ferry, et al. 1973). Few comparable studies have
Nevertheless,
been conducted in North America.
(1978) found infrared photography especially
because it indicates vitality of the organism.
some progress has been made in some parts of the
Rates of photosynthesis, respiration, and nitrogen
fixation can be measured in harvested material
following exposure to air pollution for varying
periods of time. Chlorophyll degradation can be
studied with TEM, thin layer chromatography and/or
continent.
Lecanora conizaeoides
is unanimously
according
Parmelia sulcata, and,
physodes,
Hypogymnia
regarded as
species in Europe.
the most pollution-tolerant
to some studies, Evernia prunastri are also
spectroscopy
tolerant. In increasing order of sensitivity, Parmelia
perlata, Usnea ceratina, Lobaria pulmonaria and
cellular
Parmelia saxatilis come next, the last four being
transmission
extremely sensitive to atmospheric pollution.
P. sulcata, P.caperata, P. saxatilis, U.subfloridana,
with
(Chervin
Henriksson
et al. 1968, Jacobs
et al.
useful
and
1971, Holopainen 1983).
Damage to
membranes can be measured directly with
Ahmadjian
(TEM) or
electron microscopy
the potassium
efflux method (Puckett
indirectly
et al.
most of North America.
1977) or the simpler electrolyte leakage method
(Pearson 1980, 1985; Fields and St. Clair 1984).
TEM can also be used to observe damage to plastids
the
and other organelles.
U. ceratina
and common
are widespread
and
throughout
E.prunastri is common in
Pacific Coast states and L.pulmonaria is
relatively common in mature forests of the northern
tier
of
5
indicators
listed above are either
or do not occur in North America.
for more information
See
on these
exposure to air pollution,
observation
Chapter
on sensitive species.
organelles
of
it now appears that
including
the effects
must be made.
of pollutants on lichens,
destructive
involve
The amount of material needed
sampling.
for destructive sampling must
before the transplants
be carefully
are made.
for planning is to multiply
A
estimated
rough yardstick
the number
photosynthesis
rates
and patterns, are also manifested
early.
desired by the number of sampling
dates,
plus at least 10% to cover losses due to herbivory,
1977), spectroscopy
or other causes not related to pollution.
The simplest type of evaluation consists of
measuring the survival rate of a species and
expressing this as percent survival: how many of the
transplants died showing symptoms of injury, how
many died or disappeared as the result of causes not
related to pollution, and how many survived relative
but more tedious direct chemical
vandalism,
Related
to the original
number.
morphological
changes in color and thallus thickness,
chart, available
a
of
traditional
analyses (Pearson
spectroscopy of
1987). Energy-dispersive
X-rays is showing promise in microchemical analysis
of lichens transplanted to areas near chemical
processing
plants.
LITERATURE CITED
to this are observed
We recommend using
can be used instead
and Rope
altered appearance of the apothecia or perithecia,
width of lobes, etc.
and
leakage from the cells
For evaluating accumulation of toxic elements
(Hale and Lawrey 1985, Laaksovirta and Lodenius
1979, Schwartzman et al. 1987, Steinnes and Krog
of
treatments desired by the number of
experimental
replications
observations
Some of these observations
membrane,
the plasma
have the greatest potential for revealing injury
following short time exposure at low levels, at least
in areas where sulfur dioxide is the primary
Concentric rings appear to be visibly
pollutant.
altered more rapidly following exposure to low levels
of pollution than any of the other organelles.
Other physiological and biochemical
measurements, such as chlorophyll degradation and
EVALUATION
To measure
TEM
rings and other
the concentric
measurement of electrolyte
baseline data are needed and periodic
for more
5 and 7
See Chapters
methods.
For evaluating damage to lichens following
but is now rare in reforested areas.
states,
The other European
rare
information
Ahmadjian,
Munsell
V. and J.B. Jacobs.
reestablishment of lichens
in many art textbooks and art
to aid in describing color
1982.
III.
development of Usnea strigosa.
sections of libraries,
Lab. 52: 390-399.
93
Artificial
Synthetic
Journ.
Hattori Bot.
—,
— , L. A Russell, and K. C. Hildreth. 1980. Artificial
reestablishment of lichens I. Morphological
interactions between the phycobions,
Cladonia cristatella, and Lecanora
Mycologia 72: 73-89.
I.M.
Brodo,
42:
—
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1961.
corticolous
Cellular injuries in epiphytic lichens
Nordic J. Bot.
transplanted to air polluted areas.
chtysoleuca.
Jacobs J. B., and V. Ahmadjiaa 1971.
experiments with
Kandler,
two species of Usnea.
B. J. 1973.
Coppins,
(eds.)
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Dibben, M. J. 1971.
Kershaw,
In B.
Laaksovirta,
accumulation
Athlone
Hale, M.E. Jr. and J. D. Lawrey.
lead accumulation
baltimoresis .
in
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of
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—
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C. D., R. C. Weast, and S. M. Selby. (eds)
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— and
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epiphytic
T. H. 1983.
Ultrastructural
Ohio.
physodes, growing
increases leakage of
pollution
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158: 87.
damage to cell membranes in
Development
of
Environment
S. E. Benson.
nature.
changes in
—
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a simple monitoring
Laboratory
growth
in
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and E. Henrikssoa 1981.
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Ann. Bot.
test.
19: 209-212.
Air
cell membranes in lichens II.
near a fertilizer
plant and a pulp mill in central Finland.
pollution
experiments with lichens based on distribution
lichens, Bryoria capillaris and
Hypogymnia
Fennici
1980.
Atmospheric
Holopainen,
Varying environmental factors in
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electrolytes from lichen cells.
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Chemical
Royale
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av mossors och lavars vitalitet med hjalp av infrarod
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33-40.
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des
des
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Metodundersokning
Bull. Soc.
de Belgique
Pearson, L. C. 1970.
D.L. and F. Rose. 1976. Lichens as
Pollution Monitors.
Edward Arnold Publishers,
Limited.
Londoa
1978.
La Culture
Thalles Soumis a la Pollution.
lichens: an
5-7.
113-117.
de Botanique
Tomassini.
rate
1974.
75:
Untest de la Vitalite
Nieboer, E., D. H. S. Richardson,
the lichen Pseudoparmelia
The Bryologist
16:
coal mine ecosystems in
Ohio J. Sci.
Soredies Licheniques:
Hawksworth,
Hodgman,
of
Margot, J. and J. Deslooves.
Amer. J. Bot. 71: 986-998
chlorochroa.
Mercury
1979.
Ann. Bot. Fennici
1975.
Lichen
of some heavy metals from acidic
southeastern Ohio.
SO2 on photosynthesis and carbohydrate transfer in
the two lichens: Collema ploycarpon
and Parmelia
Pearson.
K. and M. Lodenius.
surface substrates
Fields, R. D. and L. L. St. Clair 1984.
Henriksson,
4: 83-87.
J. D., and E. D. Rudolph
Lawrey,
Wiley,
and D.L. Hawksworth
Air Pollution and Lichens.
(eds.). 1973.
Press, London.
A
1969.
lichen growth chamber.
208-212.
New York.
Ferry, B.W., M.S. Baddeley,
and Aquatic
18-22 May 1987.
(France)
content of fungi in Helsinki.
5: 1-10.
Desiga
Grenoble
on
Communities
on Terrestrial
K. A. and J. W. Millbank.
Lichenologist
culture in a
Experimental
the European
controlled environment
Athlone Press.
Whole-lichen
Federer, W. T. 1955.
Ecosystems."
and D. L. Hawksworth
and Lichens,
Lichenologist
phytotroa
1968.
The "drought hypothesis."
of
Air Pollution
"Effects of
Canadian J. Bot. 46: 241-245.
W. Ferry, M. S. Baddeley,
Lichen and conifer recolonization
In Symposium of the
O. 1987.
Commission
and mycobiont in
phycobiont
7:
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Lichen growth and cities: a study on Long
Island, New York. The Bryologist 69: 427^49.
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The
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71-82.
1966.
R. R., G. E. Baker, and H. R. Hanoi.
of
II. Cladonia cristalella:
The lichen and its isolated symbionts. J. Phycol.
ultrastructure
lichens using a new technique. Ecology
The ultrastructure
4:
393-408.
mycobionts
838-841.
Chervin,
1984.
pollution
damage to
Laboratory
The Bryologist 84: 516-520.
— and G. A. Rodgers. 1982. Air pollution damage
cell membranes in lichens III. Field experiments.
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Phyton (Austria)
94
22:
329-337.
to
—
air pollution
Botanical
—
Schwartzman, D., M. Kasim, L. Stieff, and J. H.
and S. K. Rope. 1987.
Electron microscopy and
cell membrane leakage tests as aids in monitoring
with lichens.
and E. Skye. 1965.
Science
Air
pollution
Air, Soil Pollution
No. l-126c-8.
affects pattern of
Skye, E. 1958.
1977.
E. Nieboer,
—
and D. H.
Potassium efflux by lichen
New Phytol.
79:
Rope, S. K. and L. C. Pearson.
biomonitors
pollution
1990.
Ryan, B. D. 1990.
wilderness
Thompson,
Botany Baseline
J.
ecological
1969.
past.
and air quality in
areas in California.
conditions
of
Maria Curie-Sklodowska,
J. W.
Academy
of
the habitat.
Annls. Univ.
the
Will-Wolf,
S. 1980.
communities
C, 23: 131-164.
Mercury,
95
arsenic, and
complex studied
Oikos 23: 160-164.
out of Wisconsin's
Rev.
19: 20-24.
In
der biologischen
1491-1511.
Structure of corticolous
lichen
before and after exposure to emissions
from a "clean" coal-fired
Bryologist
17: 325-341.
Die Kultur von Flechten.
Handbuch
Aberhalden
Arbeitsmethodenl2:
as indicators
Lichens
1970.
Wisconsin
Tobler, F. 1939.
Univ. Arizona Dept.
Studies Series.
Lichens
1977.
by means of lichen transplants.
Lichens as air
in a semiarid environment in
Lichens
indicators of air
selenium fall-out from an industrial
The Bryologist 93: 50-61.
Idaho.
Rydzak,
Steinnes, E. and H. Krog.
125-45.
i Narkes
133-190.
Ann. Rev. Phytopathology
pollution.
thalli following exposure to aqueous sulphur
dioxide.
Lichens as biological
1979.
inverkan pa busk-
kring skifferoljeverket
Svensk Bot. Tidskr. 52:
Kvarntorp.
Water,
363-378.
32:
Luftfororeningars
ock bladlavfloran
1600-1602.
Puckett, K. J., F. D. Tomassini,
S. Richardson.
Quantitative monitoring
airborne lead pollution by a foliose lichen.
in Parmelia sulcata, a corticolous
148:
of
Johnson, Jr. 1987.
International
Congress Abstracts, Berlin:
photosynthesis
lichen.
XIV
83: 281-295.
generating station.
The
Collection and Chemical
Analysis of Lichens for
Biomonitoring
Larry L. Jackson,
Jesse Ford, and David Schwartzman
This chapter discusses the interrelated aspects of biomonitoring using
analysis of lichens. Many unique aspects of study objectives,
chemical
study design (including
design tasks, considerations,
schemes), sample collection,
sample preparation,
are required for a successful
biomonitoring
of
advantages and disadvantages
chemical
of
analysis
and sampling
and sample analysis
program are emphasized.
of
Aspects
assurance program and final contract reports are highlighted.
some examples
of
studies using chemical
INTRODUCTION
of
analysis
content
lichens to define the influence
sources
of
specific
Lichens have been used often as receptor-based
in air quality studies. Historically, they
have been used in a qualitative way, in which their
dwindling populations indicate poor air quality or
chemical contamination.
In the last few decades they
have been used increasingly in a more quantitative
variability
fashion
provide interpretable,
their chemical
The objective
content
important to examine both temporal and spatial
the
step-by-step
of
meaningful,
instructions
environmental
and defensible
data
are
not given because in any
and compromises
study, decisions
must be made based on the specific
study objectives,
the particular
budget, and the
laboratory
ecosystem,
facilities
other considerations.
the project
available,
along with numerous
In addition, this chapter is not
intended to examine those research studies where the
chemical
content
of
lichens has been used to identify
and elucidate the biogeochemical
the chemical
studies have attempted to
address only the spatial variability
in the chemical content of lichens to
for use in air quality management decisions.
This chapter highlights the issues related to both
temporal and spatial variability which must be
considered in using the chemical analysis of lichens
The interrelated aspects of such
for biomonitoring.
studies are illustrated in figure 1. Specific
variability of a few selected elements or chemical
species in a given lichen population through time
and/or space. The paucity of published chemical
data for lichens forces many of these studies to
determine a chemical "baseline" or "reference point
in time" for one or two lichen species within a
politically or geographically defined region. The
intent of such studies is typically to examine the
long-term variability of a selected suite of elements
or compounds by remeasuring the chemical content
of the lichens at some point in the future. Spatial
variability in the chemical content of lichens is
commonly a confounding problem in such studies.
Other environmental
point or
of these pollutants. Variability of chemical
constituents in lichens with time may be a
confounding problem in these studies. Hence, it is
content indicates poor
of most studies focusing on the
of lichens has been to determine
of
and to
origins
air quality.
chemical
pollutants
between natural and anthropogenic
differentiate
biomonitors
whereby
a quality
In addition,
lichens are discussed.
of
nonpoint
The
methods suitable for
common analytical
lichens are briefly discussed.
that
content or uptake
lichens—studies
96
elements by
that are much less numerous
lichens than for vascular
the chemical
processes affecting
of
plants.
for
Conceptually,
in an air quality study it is easy
or potential adverse impacts in an ecosystem.
few chemical
expensive.
species this is possible,
Expense
For
controlled
In addition,
albeit
between airborne chemical
inexpensive
how they interrelate.
STUDY OBJECTIVES
is limited.
Most environmental studies examining the
content of lichens are used for monitoring,
regardless of whether they address temporal or
spatial variability, or both. Monitoring may be used
analysis
alternative
growth characteristics,
and longevity,
used as passive collectors
of
chemical
to:
point in
baseline against
which future comparisons may be made
to determine long-term trends,
1) study
they may be
However,
the
living chemical
sampling devices are fraught with difficulties that
practicalities
using lichens
must be addressed to provide
interpretable
2) study the extent
release or spill,
and
results.
The major aspects
interrelated,
as
meaningful
as shown
of
a biomonitoring
in figure
1.
at a reference
of
a
a
single chemical
3) determine the region of influence
and nonpoint pollution sources,
study are
For example,
conditions
time, to determine
over time periods
extending beyond just a few years.
aspects of the overall
influence each part of a
biomonitoring program must give
appropriate attention to all of these study aspects and
of lichens offers a relatively
to air sampling in which air
contaminants can be indirectly measured. Many
lichen species obtain much of their nutrients directly
from the air and rainfall, thus serving as integrators
of air quality. Because of their ubiquitousness,
The chemical
the planning
study. A successful
concentrations
and adverse impacts on plant communities
the
objectives]
Sample
•
•
•
Collection
QA/QCl
Preparation
Chemical Analysis
Data Analysis
Figure 1. — Interrelated
of point
4) assure that pollutant levels are within
acceptable levels, and
study objectives directly influence the study design
used, the species collected, the types of chemical
STUDY
DESIGN
to be
quality assurance program
of measurement sites to only a handful within a large
geographic region. For other potential airborne
constituents, collection and measurement may be
difficult, if not impossible, with current chemical
In addition, our understanding of the
technology.
relationship
by the nature of the population
studied in time and space and by the project budget.
a
restricts the number
frequently
In turn, each of these aspects is
assurance program.
and rainfall to define existing conditions
particulates,
and the nature of the quality
analysis performed,
to
suggest that one should analyze the air, airborne
aspects of biomonitoring using chemical analysis of lichens.
97
•
5) determine those chemical or biological
parameters that are measurable and/or
space.
realities
of
sample collection,
analysis,
In developing
of
of
an environmental
Aberrant
or
with future collections
chemical
when
are to be made.
may artificially induce
seasonal conditions
negate
emission
are important
trends by introducing
uncontrolled
In addition, the length of time required
for sample collection must be considered to avoid
uncontrolled temporal variability. For example, the
the ecological and
Understand
of
setting
the target
chemical
Define the samples to be collected and the
field measurements to be made.
Develop a quality assurance program for
sample collection, chemical analysis, data
after a period of rain may be significantly different.
(i.e., qualitative
uncertainty
may influence
Numerous
or
in collection and analysis of
estimate potential
to determine
scales
and
of variability or
Preliminary sampling should
spatial trends.
be done using the same collection and
analysis
techniques
monitoring
program
as the primary
to produce valid
Develop
based field sampling
Re-evaluate
data quality
preliminary
sampling
and statistical
Execute
protocols
existing
tests to be
analysis
objectives
of
a study design.
are important as
and anthropogenic
patterns for pollutants,
monitoring
Characteristics
Ideally
schemes.
the sampling
of
pollution
and
systems must be taken into
of
the lichen
target population
also be evaluated in creating the study design.
based on
results and sampling
the study according
or natural
account.
on chemical analysis results.
performed
the viability
sources, dispersion
statistically
in
human-caused
or development,
spatial considerations
the study area to natural
conclusions.
plan and define statistical
Potential
For example, ecosystem, physiographic, and
influence
geological boundaries all may significantly
the chemical content of lichens and the magnitude of
the heterogeneity observed in a geographic region.
Differences between micro- and macro-habitats may
artificially introduce spatial trends unrelated to
potential pollution sources. Political boundaries must
be considered with respect to permission to sample
In addition, the relationship of
and habitat stability.
Review literature and/or perform
element concentrations
studies.
well.
samples).
sampling
before and
a habitat must also be examined
long-term
changes such as logging
and quantitative
quantifiable
samples collected
changes caused by events such as fires or hurricanes,
levels of variability
preliminary
of
The stability
designing
and define the data quality
statements regarding
content
of
population.
objectives
•
weather such as wet
variability.
analysis,
•
of
These considerations
comparisons
study are (after Gilbert 1987):
the target population in time and
environmental
•
plan, temporal
seasonal aspects of plant
and seasonal pollutant
herbivore activity,
patterns.
space.
•
the sampling
include
natural cycles
physiology,
and must meet the study objectives.
The basic design tasks
•
have been
and dry seasons or seasonal shifts in wind patterns,
good study design enumerates a series
responsibilities
•
objectives.
the study objectives
Selection of Target Population
Design Tasks
Define
the chemical
Design Considerations
considerations
•
in
results and evaluate them
and budgetary
STUDY DESIGN
monitoring
if
Determine
constraints.
A
the uncertainty
met.
must be tempered with the
Study objectives
results and
with respect to the data quality
•
to be sampled in both time and
target population
Determine
and statistical
to be tested and the
the hypotheses
analysis
trends.
•
should be clearly defined and
Study objectives
the chemical
determine relevant temporal and spatial
changing temporally or spatially in order
to define future monitoring programs.
must consider
Evaluate
the
sampling
to written
•
sensitive to the pollutants
with
and quality
assurance plans.
individual lichen species chosen for
are:
and abundant)
98
of
interest, but
a stable, robust population
(i.e. healthy
must
•
with measurable
of interest
common species present in monotypic
of
concentrations
•
samples, and although they are generally
good passive collectors
stands on the same substrate throughout
judgmental
study area
•
easily identified in the field
easily collected
may be as simple as
haphazard grab samples, or quite
and based upon known dispersion
sophisticated
and cleaned in sufficient
patterns and plant uptake
of
quantity
sufficiently abundant that repeated sampling
not seriously impair the viability of
local populations
species with known physiological processes
inferences
conclusions.
such as growth
of
rate
are dependent
of
and prejudices
and nutrient uptake, and
In
a few grab samples,
A
purposes.
In addition, the lichen substrate should have low
and lead to more interpretable
results, even for orientation
of
the elements and/or compounds
Simple random sampling,
interest.
haphazard sampling,
The considerations
listed above are used to
To have
time and space.
other practical
Logistical considerations
ability
to precisely
sample site, ease
of
sampling
design.
of
integral part
particular
sampling
and preparing
or other parameters.
equipment
required, and
ability
These logistical
evaluating
sampling
issues become an
the cost-effectiveness
of
However,
to a priori pre-select
are more homogeneous
affect the
compounds
of
within
design.
a grid,
Whether
or spatial issues, population
concentration
within
of
generally
determined.
sub-populations
involves
The terms bias, the amount
sampling
are to each other, are commonly
analytical
techniques.
used to describe sampling
Sampling
categories:
sampling
used
They may also be
designs.
and random sampling.
legally
defensible
estimate the
design, and
or patterns in the data may
upon the frequency
of
sampling
studies where the objective is a map of a population
such as an element concentration, the
of a particular map is referred to as
An unstable map produced from a
systematic sampling scheme with insufficient sample
density may produce misleading or incorrect results.
The
reproducibility
designs can both be probability
than judgmental
if
characteristic,
map stability.
based, and hence have less relative bias and are more
However,
Systematic
in time or space. The temporal and spatial
interaction of the population heterogeneity, and trends
with the sampling density control the reproducibility
or precision of the sampling design. For spatial
fall into three
judgmental or haphazard sampling,
latter two sampling
error in a systematic
be missed depending
designs generally
systematic sampling,
at points
or by some
These designs are
used for evaluating trends. However,
unsuspected periodicity
true value, and precision, how close repeated
determinations
which
or probability based
it may be difficult to accurately
are
how close the measured amount is to the
to describe
means
this depends on the
along a line or gradient,
most frequently
variability
measured above or below the true value,
consistently
accuracy,
of
a subset may
population
they use a random starting point.
parameters such as
means and magnitude
a
for the elements or
designs can be judgmental
is focused on temporal
a study objective
is
into homogeneous
other set pattern in space or time.
Selection of Sampling Design
of
the target
interest.
Systematic sampling
a
of
Stratification
more accurate estimates
collecting,
all ultimately
of
random sampling
Simple
subsets with random sampling
provide
same as
the
or spatial patterns exist in
heterogeneous target population
such as site accessibility,
convenience
temporal
the target population.
study design,
must be weighed.
of finding,
if
not suitable
locate and re-find an individual
the target species,
other issues
a workable
considerations
which is not
offers the least relative bias
is assumed.
population
and to define it in
study
purposes.
schemes, but homogeneity
the sampling
that will
the specific target population
identify
become the sampled population,
may be made at
no inferences
probability -based pilot study may be
more effective
of
of
case, as in the collection
all and the results may be used only for orientation
with previously measured chemical content
from this study area and other regions.
concentrations
Also, it is difficult
bias or accuracy
the worst
but
upon the knowledge
entirely
the collector.
to estimate the statistical
In
pollutant.
a specific
general, these designs may be cost effective,
will
•
a few
collecting
•
•
sampling.
Judgmental sampling
the
preferable,
any more accurate than
they are not necessarily
the pollutants
sampling.
they may require a larger number of
The various
appropriate
advantages and disadvantages,
calculation
and their combinations,
99
aspects
of
each type
are discussed
and the
of
design
in numerous
estimates of the variance
texts such as those by Gilbert (1987) and Keith
(1988, 1991). More specific texts are also available
on time series analysis
or spatial statistics such
kriging and contouring
(Davis 1986, Isaaks and
Srivastava
Use
in designing
must be considered
results.
evaluating
Whereas
will
assume that the data
distribution of
frequently
about
study.
data in plants are skewed and must be
A
in order to approximate normality.
A "Chi
logarithmic transformation is common.
Square" test or the non-parametric Kolmogorov-
log-normal distribution
The various
or other
and its variance
the
or geometric,
if
example,
are estimated for chemical
of
due to analytical
and to heterogeneity
Ecological characterization
sources
analysis
and sampling
population
without
parameters may be erroneously
a proper
estimate
of
the analytical
baseline concentration
contributor
ranges
to the variance
if
so
of
interpreted
•
of
Estimation
and representativeness.
of
range of chemical
and magnitude
These tasks are necessary parts
the largest
of variability.
of
a primary
regardless of whether or not an orientation
is the analytical
performed.
be performed
protocols
to
orientation
of the analytical measurement and/or
collection of samples. Unbalancing an analysis of
primary
approach to providing
100
study,
study is
In general, the orientation study should
using the same collection and analysis
be used in the primary study. The
study results may dictate that changes be
made in collection or analysis
must take into account
design is a common
Analytical methods evaluation for required
sample size, detection limits, accuracy,
concentration
replication
variance
between funding agency (data
(samplers), and
Coordination
precision,
For
error.
and it contributes
a study design
of quality assurance program
for collection, chemical analysis, and data
analysis which includes quality control
Development
chemists.
the total variance.
Thus,
effort to
users), contractor/researcher
more than 50% of
This situation is not uncommon
when an element concentration approaches the lower
limit of detection of the analytical method.
measurement,
of
samples.
•
it is difficult to justify the calculation
example,
level
accessibility,
study design and implementation
•
by the analytical
Estimates
Evaluation of logistical considerations,
with potential primary study
It is especially
can be estimated.
region,
design.
that the study design include replication
measurement
the
species for
compatible
•
to
Klusman 1985).
No analysis of variance is possible without
important
lichen
Orientation
Hierarchical or nested
been used in this fashion
that the error contributed
review.
of
•
attributed to the various
measurements.
literature
•
spatial scales (Leone et
al. 1968, Miesch 1976, Severson and Tidball 1979,
of
study, which
the field and
find and collect target species.
Necessary collection permits.
error,
estimate variability at various
replication
comprehensive
convenience,
within a sample site and
Analysis of variance may be
of heterogeneity.
of variance has
of
collection.
•
between sample sites.
used to estimate the variance
study.
work, include:
•
is the sum of the variability
measurement
priori
an orientation
habitats, and potential
from a
samples is collected
region, the total variance
of
A
data.
a series
aspects
•
The total variance is the sum of
For
variances for individual components.
concentration
constraints
orientation
research study that is little more
orientation
should represent about 10-15%
analytical
the mean, either arithmetic
a primary
than an expensive
distributions.
Typically,
and a
Instead, the experience
commonly
test may be used to evaluate the goodness
a
project budgetary
do not permit a reasonable
of a researcher is assumed to suffice.
The result of such policies for studies of chemical
concentrations in plants or lichens is all too
transformed
of fit for
However,
primary
knowledge
Frequently,
concentration
Smirnov
and cost-effective
a successful
study design.
distributed,
in
study is invaluable
good orientation
producing
statistics
parametric
be normally
the data.
A
a study and in
statistics make no assumption
non-parametric
the
The Orientation Study
statistics
versus nonparametric
parametric
scheme
sampling
(Miesch 1976).
as
1989).
of
while
components
a cost effective
maintaining
research study.
procedures
for
the
The Primary Research Study
supplementary
The primary study includes refinements of the
tasks listed above, based on the results of the
samples at additional sites or times, resampling
specific sites, or reanalyzing specific samples.
orientation
An important facet of this process
study.
is evaluating
the data quality
sampling
may involve collecting
Supplementary sampling is usually needed to verify
conclusions, clarify problems such as outlying
needed compared to the
critical
quality attainable as estimated from the orientation
values, estimate parts of residual variability not
study. This evaluation must include budgetary and
resource constraints, with the potential outcome being
a need for additional resources or a reduction in the
quality or representativeness of the data.
During the primary study, strict control must be
maintained over levels of stratification in both the
To maximize the
space and time domains.
interpretability of the chemical concentrations in
associated with laboratory
that are representative, that will meet the study
lichens, care must be taken to clearly
objectives,
identify
sample plan the biological and physical
geographical
used.
if
levels,
stratification
of
the
variation,
or explore
results.
SAMPLE COLLECTION
The goal of sample collection is to obtain samples
in the
stratification
is
collected
uniformly
and that are
to the laboratory),
transportation
stratification
with
or during
that are minimally contaminated
substrate (by the collection procedures,
or
Spatial factors that may influence
facets
unanticipated
throughout
of
the
as opposed
to
the duration
Collection of single species
include such things as macro- and micro-habitats,
study.
host/substrate for lichen (e.g., fir versus aspen, or
tree versus soil), physical orientation (e.g. lower
Taxonomic
mixed-species specimens is preferred.
vouchers should be collected to allow for expert
and geological
physiographical
verification,
trunk, or site aspect),
canopy, south-facing
to pollution sources.
Temporal
factors, such as
The sampling
estimates
analytical
of
must be maintained
homogeneous,
sufficient
is commonly
of known or
site to quantify
geochemical
the design by inappropriate
aspect of the primary
will
be used to test them.
part of this is evaluating
its potential biochemical
or
influence.
analysis
One
expected dictates both the sample size and the nature
of
statistical software.
the contamination
Contamination
results should be designed to
for additional data and avoid
potential problems in the reporting of results. Sample
from sampling
schemes must also be
trace metal analysis
sampling
10-15% of the field and analytical
of
lichens,
sample tools should
of Teflon®, plastic, Teflon-coated metal, or
stainless steel (in descending order of preference) to
minimize metal contamination. Plastic tools may be
inappropriate for organic pollutant studies, however.
Oils or greases used to lubricate sample tools may
at least the
that would represent
work.
tools or storage
or that occurs during transportation
(fumes and particulates) may obscure the true
chemical concentration trends. For example, for
be made
potential for additional
that must be used in
containers,
developed at this point.
Supplementary Sampling
A study design should incorporate
control
both the sample collection and analysis.
the need
and tracking
must be
for the elements or chemical compounds
of interest. The level of environmental pollutant
At this stage, the database for field measurements
identification
It may also
sample substrate at each
appropriate
whether the design created
can be analyzed using existing
identify
the
The sample collection and handling
study design is
to explicitly define the working hypotheses and the
and chemical
needs.
should be uniform from
Sample Handling
stratification.
statistical tests that
required to meet the laboratory
suspected variance while at the
same time not biasing
An additional
and to allow for
Compositing of lichens
results.
be necessary to collect
between controlling for major
the
to obtain a
site to site and should be documented.
In
etc.
identify
sample, to provide
representative
The nature of compositing
error and the
sampling
form, biomass,
sample for analysis,
reanalysis to verify
design, a reasonable balance
creating the sampling
sources
the
measurement error must be included.
growth
plan should clearly
amount of sample to be collected
pollution events, phenological seasons, growth stages,
and intra- or inter-annual climatic differences, may
influence both initial sampling and re-sampling plans.
In general, the better control maintained over
stratification levels and the fewer the total number of
levels, the easier study results will be to interpret. In
addition,
for archival purposes, and to address
questions about
setting, and proximity
The
101
easily contaminate
of contamination
Other potential
samples.
include
sources
mercury. Freezing
wood which may
objects (e.g., Wolmanized®
not common.
high levels of As, Cr, and Cu), personal
introduce
samples immediately
Storage temperatures should be at
-20o C or below to minimize
gear such as gloves and insect sprays, and activities
chlorinated
For organic pollutant studies,
insect repellents or other lotions or sprays should not
be used in the field unless it can be demonstrated
storage on the order
such as smoking.
that
the relevant chemicals
interference
analytical
will
upon
collection is possible using liquid nitrogen, but it is
treated wooden
chemically
or
hydrocarbons
of
C
if
long-term
years is planned.
Collection Site
Sample
neither contribute an
of volatile
loss
at -80o
The sample collection site should meet the criteria
nor serve as a contaminant
Finally, if the
concentrations of the elements or compounds of
interest are extremely low, special care must be taken
target lichen species, type of habitat, geographic
to avoid
sources (e.g., roads), and stability
source for target compounds.
contaminating
the
levels in the laboratory
particularly
important
This is
setting.
for volatile species such
compounds
available
bottles or bags, cloth bags, glass bottles, or
Plastic
for
locate a site is critical
to accurately
some studies in which spatial statistics are used.
No
one container is suitable for all studies because of
contamination
are common
possibilities,
sample containers.
general sample location
topographic
The specific sample storage container must
be selected to minimize contamination
from the
container itself (such as the potential addition of
sulfur from Kraft paper bags or plasticizers from
plastic bags or bottles that interfere with organic
determinations) or during transportation (such as
carrying cloth bags in the back of an open truck on
dusty road).
The nature
the use
of
with
grade solutions.
of
Containers
In addition,
and/or the container
invite vandalism.
positioned
Metal markers
of
used,
are commonly
such material or
such that they would potentially
In most studies, the field notes for
the chemical
field may be beneficial
transportation
time to the laboratory
contamination
in
laboratory
concern.
if
protected with an appropriate
of
diligence.
during the drying
for their own purposes, it should be kept in mind
Air-drying in
that these
the
notes may be used by other researchers for
constituents
thoroughness
temperatures do not
exceed normal ambient temperatures.
effects.
Hence the degree of
and legibility
In addition, referring
Refrigeration
or freezing of samples is generally not required and
may even have deleterious
degree
Whereas most researchers take adequate field notes
is long or
the agency in the future.
as long as drying
a site are as
valuable as the samples and should be recorded and
field should not induce excessive loss of volatile
of
of
Field Documentation
interest.
the
but
must not promote deterioration
volatilization or transformation
process is a potential
Markers
on the environment,
contaminate future samples.
the sample handling
sample (such as by mold formation or rotting
when wet samples are stored in plastic bags) or
the
be consistent
required by the study
by the property manager.
but they should not be made
and
the
Air-drying in
the site should
permanency
should be locatable in the future and should not
should not permit cross-contamination
between samples.
of
of
the degree
and allowed
a
degrees
such as
System, Loran-C, or aerial photography.
Permanent marking
the
using appropriate
Other
compass and chain, satellite Global
should not be obtrusive
of
scale such as the
series.
may be required,
accuracy and precision
Positioning
study may dictate pre-cleaning
the sample containers with acid or solvent washes
compounds
Survey Quadrangle
locating techniques and devices with varying
of
The
should be recorded on a
map at the appropriate
US Geological
cost, and transportability
issues.
freezing
the study and the
studies in which the site must be resampled, and for
paper envelopes
of
of
habitat.
should be
technology.
The ability
biphenyls
of
the site location
consistent with the objectives
as
(PCBs)).
handling
of
contamination
to confounding
proximity
Determination
(e.g.,
and polychlorinated
hexachlorobenzene
location,
samples due to high
background
mercury and organic
dictated by the study design, such as presence of
to the
of
the notes is important.
field notes may be
only way to resolve problems
the
in data interpretation.
The field notes for all sites should be uniform,
However,
samples may be required for long-term
preprinted field data sheets should be used.
storage or to impede the degradation or volatilization
of organic pollutants or of volatile elements such as
notes have several aspects in common.
generally
102
should include:
They
and
All field
•
•
site identifier,
location
information
coordinates,
hand-drawn
topographic
sheet name),
general information
Thus, in the following section, we
elements.
(e.g., site
site map,
some
highlight
•
habitat or community
sample descriptions,
field measurements,
miscellaneous
problems
sample, particle size reduction,
on collection
comments
Cleaning and Washing
numbers,
•
date
•
names of sample collectors, signature of
field note transcriber.
an encoded sample identifier scheme is used
a simple
foreign
sequential numbering
picking
should be made of
and sample collection
•
SAMPLE ANALYSIS
for
the chemical
Whereas chemical
analysis of plants, and
of
plants and lichens has
been used for some time for pollution monitoring,
majority
of published
nutrient levels (Westerman
Jones et al. 1991, Jones 1991). Very few
Will
Hand
may be done to
in the lichen?
a washing
significant
procedure
portion
of
remove a
the surficial
contamination?
the
•
methods on plant analysis
address agricultural
a microscope
Is surficial contamination present?
Is the concentration of the elements or
compounds of interest in the contamination
significant with respect to their
concentration
•
have not been standardized.
analysis
of
Several issues must be considered:
•
Protocols
the aid
plants to remove such material is controversial.
and compositing methods. Where possible, sample
site identifiers should be included in photographs.
lichens in particular,
from the lichens themselves.
with
difficult, it is tedious and does not remove surface
dust. Most plant surfaces are contaminated with
windblown dust, fly ash, automotive particulates,
agricultural sprays, or a variety of other airborne
The efficacy of washing lichens or
particulates.
general site habitat, lichen growth habit and nature of
morphology,
is to remove
elements and/or
remove adhering substrate such as bark or soil
collectors.
substrate, thallus
lichens
leaching
particles, extraneous plant species, and dead or
decaying tissue. Whereas hand cleaning is not
scheme, it
on the field data sheet to
confusion and mislabeling by the field
documentation
matter without
compounds
should be preprinted
Photographic
in cleaning
The objective
and time, and
minimize
homogeneous
of organic matter.
or deviations from sampling
photograph
of
and destruction
subsampling,
•
If
that
prior to chemical analysis of
lichens are cleaning, washing, and drying of the raw
must be considered
design,
instead
methods in
concern.
The various aspects of sample preparation
samples collected,
sample numbers),
•
of
Lichen Sample Preparation
description,
including vouchers and identifiers (field
•
analytical
the primary
such as weather or
other pertinent temporal conditions,
•
of
current use and the issues
Will
a washing
amounts
1990,
of
procedure
leach significant
of
the elements or compounds
interest from the lichen itself?
schemes have been used to
analysis procedures
Numerous
for lichens. In general, the analysis methods and
issues of concern for lichens are the same as for
clean plants:
agricultural
dilute acid washes (Jones and Case 1990, Little 1973,
Saiki and Maeda 1982). Goyal and Seaward (1981),
publications
describe
chemical
washing
tap or distilled water rinses (with or
without ultrasonication),
mild detergent washes, and
crops.
Like every other aspect of lichen biomonitoring,
the choice of preparation and analysis methods
and Hale (1981), and Jackson et al. (1985)
observed leaching of metals and sulfur from lichens,
but it is difficult to tell whether the decrease in
Lawrey
depends upon the study objectives, the concentration
of elements in the lichens, and budgetary constraints.
the prejudices of the researcher and
facilities available also dictate the
Quite frequently,
element concentration
the laboratory
(i.e., with additional
sample preparation
suite
of
and analytical
elements determined,
methods used, the
washes) is due to more effective removal
contaminants or leaching from the lichen.
and the determination
limits for the elements.
can be defined
as the
with more aggressive cleaning
cycles or more vigorous
washing
Realistically, no one method
only suitable method for most
of
Little
(1973) found that water washing removed the
majority of the surficial deposits from foliar samples.
but the total amount removed was element
103
Thus,
dependent.
deciding
of
of
the nature
upon a washing
scheme on the final chemical
any washing
particle size reduction
used, including
methods have been
hand grinding,
kitchen
or industrial
blenders, Wiley mills, ball mills, and cyclone mills.
The choice depends upon the amount of material, its
results
when providing the
must be considered
environmental
Various
The influence
procedure.
and/or loss of sample material.
contamination
samples and their
must be critically examined before
contamination
interpretation.
of
the methods
morphology,
chemical
analysis,
sample size required, and the chemical
Sample Drying
determined.
changes in the
Drying the samples minimizes
tissue once the sample is collected,
and provides a
uniform weight basis for analysis.
Jones and Case
Frequently,
analysis.
60o C are generally
grinding.
in
but that excessive weight loss
occurs at temperatures
above 100o C.
the National Bureau
recommended
formerly
drying
materials (SRMs), such as
for
grinding
(NIST,
materials,
that the new botanical
such as
SRM
of
lichen thallus and
the
parts also influence
the different
method and the range
of
the
sizes that
particle
source
of
and analysis
variability.
The standard bench model Wiley mill typically
Citrus Leaves,
of 2mm (-10 mesh), 1mm
(-18 mesh), or 0.5mm (-35 mesh). Jones and Case
(1990) suggest that -20 mesh material (0.85mm) is
in air in an oven at 85o C or vacuum
Room temperature for 24 hours. They now
recommend
of
The morphology
may introduce an additional
2 hours
drying at
grinder must
mechanical
(Gough et al. 1988a).
Segregation during sample handling
Standards, NBS)
standard reference
1572
the
are produced
of
SRM
of
The choice
the toughness
The National
Institute for Standards and Technology
previously
lichen material
take into account the amount of material lost during
required to stop enzymatic
a sample,
of
only a small amount
Thus it must be conserved during
grinding process to have sufficient sample for
is collected.
(1990) reviewed various aspects of drying plant
samples. They point out that temperatures above
activity
the
species to be
produces particle sizes
reference
for analysis of sample aliquots of 0.5g.
Grinding to excessively small particle sizes may
1547 Peach Leaves, be dried
sufficient
for 120 hours at room temperature in a desiccator
with fresh anhydrous magnesium perchlorate or
freezed dried for 24 hours at -5o C at a pressure of
13.3 Pa. The certificate for analysis for the new
elements such as Al, Co, Cu, Cr, Fe, Mo, Ni, Si, W,
standards now states that "vacuum
and Zn.
temperature and oven drying
have resulted in excessive
drying at room
at elevated temperatures
weight
drying
of
The use
of
For standard
and the heat
some organic
is not particularly
critical
elements.
1988a,b).
disrupts cells (Spittler
These conditions
the
do not drastically
temperatures
(generally
particularly
appropriate
chlorinated
hydrocarbons
higher temperatures.
regions.
exceed
made by determining
organic extractions
Low drying
submission
to obtain an estimate
of
precision.
"coning and quartering"
moisture on a separate
should be done.
Simply
inappropriate
and may accentuate the chemical
heterogeneity
for the different particle sizes.
a sample by "coning
and quartering,"
the
entire ground sample is poured onto a sheet of paper.
in grinding
This step is
samples are often split prior to
to the laboratory
pouring a little sample into two or more bottles is
Sample Grinding
size.
should quickly follow grinding.
basis can be
To split
particle
may be lost
action after grinding
and Bourke 1988). Therefore,
on un-dried
subsample.
The objective
compounds
To minimize between-sample
heterogeneity, a "Jones" alternating-chute or riffle
type splitter should be used. At the very minimum,
analytical
or mercury might be lost at
In cases where volatile species
is to obtain a homogeneous
organic
Following grinding,
less than 35o C) would seem
when volatile species such as
may be lost, analyses may be performed
material, and corrections to a dry-weight
However,
and analysis
for most inorganic
or converted due to enzymatic
to which samples may be exposed in
field in many geographic
a
species.
An alternative method is to dry samples in air in
an oven at 40o C for 48 hours (Gough et al.
the conditions
and provide
The length of time between grinding
samples this may be impractical.
with
In addition, excessive grinding
produced may potentially volatilize
suitable particle size.
in a desiccator or freeze
a sample
agate ball mills or shatter boxes
may reduce the metal contamination
is reasonable, but for large numbers or large
quantities
metal contamination.
grinders may contaminate
losses and
therefore are not recommended."
reference materials drying
introduce additional
Mechanical
a plant
or lichen sample
The sample is mixed by lifting the corners
sample
of
paper and rolling the sample over itself.
a prime
uniform
source of
thoroughly
104
mixing
the sample,
of
the
After
the cone is flattened
out and divided
into quarters.
sample in opposing
process
of
mixing,
to another container.
repeated until all residual
is
sample is divided between
the amount
preparation
•
Ashing of Samples
Prior to most inorganic
the organic
chemical
portion
of
•
analysis
This is generally done by wet
with a mixture of acids such as HNO3,
HCIO4, or H2SO4, or a high temperature dry
is performed. The choice of wet or
oxidation/ashing
dry oxidation is influenced by the nature of the
analytical method and the volatility of the chemical
species. For example, in acid digestions boron is
typically volatilized, whereas in dry ashing As, Hg,
•
accuracy and precision
•
interferences
•
the number
other elements may be lost.
procedures for wet oxidation
and frequently
•
the
method
of elements simultaneously
of
availability
instrumentation,
analysis
of most modern instrumental
The principles
have been reviewed
analysis techniques
by Skoog
and West (1980). In addition, Watson and Isaac
(1990) have briefly discussed the principles,
The
advantages, disadvantages,
techniques
many analytical
for plant analysis.
ashing procedures are also varied
of
of
time, and cost
are many and varied
depend upon the sophistication
of
determined
(Jones and Case 1990) and more and more include
the use of microwave ovens and closed vessels.
High-temperature
range and limit
measurable concentration
detection
the sample must
oxidation
S, and various
or total
partial (i.e., extractable)
determination
be destroyed.
Se,
many factors
of sample required (typically,
1-10 g, or more)
• destructive or nondestructive
sample
•
the two containers.
procedures,
a method,
must be considered:
The
and transferring
quartering,
In choosing
compounds.
container and the material in the other two opposing
quarters is transferred
of many organic
is also true for the determination
of
The majority
quarters is transferred to one
of
determination
the
for
and sample preparation
with regard to their
Jones (1991) has reviewed
micro-nutrients in plants.
use
the
ashing furnace used.
Samples are generally ashed in muffle furnaces at
about 500o C for 4 hours or more. Generally the
Atomic Spectroscopy
temperature of the furnace is ramped up to
450-500o C over several hours and then held at that
spectroscopy
used for inorganic
temperature for 4-8 hours or more.
introduction
ashing furnaces with variable
rates,
holding
common
Atomic emission, absorption,
Programmable
provide
analysis
They have added a degree of
in temperature control and precision
unattainable in sample ashing. Jones
Flame
use.
AAS
such as Ca, Cu, Fe,
previously
others.
time-consuming
crucibles
sample are commonly
Material from
contamination.
the walls
of
relatively
of metals in plants.
K, Mg, Mn, Na,
few interferences,
region is usually
elements
and Zn, among
of
but the linear calibration
Detection
furnace may be a major source of inorganic
small amounts
contamination
furnace (flameless) AAS for elements such as Cd,
Co, Cr, Mo, Ni, and Pb. Other flameless AAS
during
the ashing procedure.
Numerous
instrumental
the determination
with
of
analytical
sample are attainable using graphite
techniques routinely used include cold-vapor
Hg and hydride-generation AAS for the
Chemical Analysis Methods
methods exist for
hydride-forming
most metals in lichens, each
Flameless
AAS
AAS for
elements, As, Bi, Sb, Se, and Te.
techniques
typically have detection
limits in the sample in the ng g"
of advantages and disadvantages.
The choice of methods for the determination of Hg,
semi-metals such as As and Se, and the nonmetals
such as B, S, and halogens are more restricted. This
a multitude
a
limits in the
in the g g" range. Better
factor of 10-1000 with very
small.
detection limits by a
the muffle
(AAS)
Each element is determined individually,
In general, there are
procedure.
(1991) outlined a representative ashing procedure.
Quartz or porcelain dishes or crucibles are typically
used as ashing containers. Quartz or "Vycor"
less sample
Since the
spectroscopy
has been used to determine
sophistication
contribute
plants.
it has been the most widely used
method for the determination
times, and cooling stages are now in
generally
of
of atomic absorption
by Walsh in 1955,
temperate ramping
and fluorescence
the basis for most techniques
range.
Historically, arc, spark, or flame emission sources
have been used in photographic or direct-reading
spectrographs for the determination of metals. These
emission
105
techniques
and many
AAS
methods have
limits
aspects
of
sample digestion
the
limit information
4
g-1
8
4
4
g
'
g
'
2
g
'
*
g-1
4
8
g-
j
j
|
U
;
V
j
Y
;
j
4
8
4
g
g
'
'
g
g-
|
as well
Mass spectral
as
routinely
determined
of some elements
ICP-AES or AAS
by
X-ray fluorescence spectroscopy (XRF) with
or energy dispersive detector
also
technique used for the determination of metals and
Detection
some nonmetals.
ICP-AES
dispersive
Energy
XRF
used than wave-length
in plant analysis.
as
are prepared nondestructively
for ICP-AES,
XRF (EDXRF)
dispersive
more commonly
probably
Samples frequently
pressed powder,
and elements in atomic number from sodium through
all other methods, to verify whether the
for the raw plant material,
ashed material, or digested material in solution.
The
on the order of
precision of ICP-AES techniques
uranium may be determined.
5-10%
problems with this technique and may affect both
is
standard deviation
is
relative
concentration
well above
when the element
the detection
limit.
accuracy and precision.
Solid-sampling techniques have been used with
ICP-AES, such as slurry-nebulization or laser
ablation, but they are not in routine use, especially
for plants. Whereas ICP-AES has commonly
replaced AAS for multi-element determinations,
be determined
not replaced flameless
AAS
heterogeneity
and
standards pose distinct
The rapid nondestructive
analysis and the large number
of
with EDXRF make
elements that may
an excellent
screening technique.
Radiochemical Techniques
it
has
EDXRF, sample
of calibration
matrix matching
detection
range may be
it
attained with
Whereas
g"
limits in the sample in the
g
as
detection limit given
is
as
well
detection
more
ICP-AES,
information.
wave-length
upon various
and the
distribution
becoming
offers better detection
techniques.
over
Note that caution should be used in
instrumentation.
reviewing
that are
1
may vary widely depending
ICP-MS
albeit expensive.
overlaps preclude the determination
is
limits typical
lists detection
Zn ^g
sensitive detector.
isotopic
Table
concentration).
of ICP-AES analysis of
plant ash that has been acid digested.
w
Yb no
g-1
200
a
a
several orders of magnitude
M
limits for some elements than
of
wider dynamic
range (i.e., linear calibration
/yg
is
as
common,
ICP-AES techniques usually
of detection than AAS
but they have
techniques,
Th pq g°
80
has been coupled with mass
technique with the
(MS) to provide
advantages of ICP as an atomization source and MS
have similar or better limits
concentration
Ta uq
spectrometry
Simultaneous
the same number
20
1
a
elements determined.
for
g
In recent times ICP
atomic
In this technique,
require less sample than
generally
instruments
'
m
Sr j/g
4
Pb //g
8
Ni uq
8
Nd //g
'
Nb^gg-'
Sn
4
1
is
sequential
j
'
Sc uq g
a
instruments
OT1
Li
Mo /A) fl"'
used as the atomic emission
or sequentially.
g-1
Mn (jg
source and many elements may be determined either
simultaneously
|
8
Detection
Limit
i
a
coupled
(ICP-AES).
8
ml solution)
I Element
It
an argon plasma
is
spectroscopy
^j
a
been replaced with inductively
emission
Eu f/Q
4
20
La uq
4
g"1
Cu pa
g1
8
g
As vq
'
Ag //g
Cr //g g-'
How
2
0.01
Co {/q
g"'
Ti%
Cepgg1
1
0.01
P
%
1
0.01
i
Na %
Cd fjg g"1
j
0.1
2
20
Bi hq
I
Mg %
'
j
0.1
; Be vq g
2
!|
%
Ba //g g°
i
0.1
K
Fe %
|
Ga fjg g-1
16
2
0.1
Ca %
'
| Au f/Q g
0.1
(0.1 g plant ash/10
Detection
Limit
Element
2
I Al %
Detection
Limit
j Element
4
Detection
Limit
Element
|
limits for the analysis of plant ash by ICP-AES
(P. Briggs, USGS, personal communication).
detection
Example
g
"'
—
g
"'
1.
a
Table
techniques where
Numerous
lower detection limits are required, or for elements
such as Hg, As, and Se.
be used
radiochemical
for plant analysis.
analysis
techniques may
Isotope dilution
techniques have been used, though instrumental
106
neutron activation
(INAA)
analysis
In INAA,
used today.
commonly
is more
with gamma-ray
analyzed
such as chlorinated
for many trace elements is measured
in the sample
in
are
Typically, detection limits
range for most of the
various
inter-element
provide
reasonably
elements; however,
times
analysis
of
preparation
dichloromethane
determined.
of
type
of
conditions,
chromatography
(GC-ECD)
several months are more common.
number
exist in the U.S.
confined
with ICP for
In addition,
of
Measurement
of
elements.
pollutants.
The choice between the two methods is
and other organic
GC-ECD
limits and
is a sensitive
method compared
inexpensive
and
to GC-MS.
for hydrocarbon
of PCBs
hydrocarbons
with eluents of differing polarities may
In GC-ECD analysis, peak identification
is accomplished
for uniquely distinguishing the
of sulfur to an ecosystem from various
from most other chlorinated
by comparison
with chromatograms
/S
sequential
fractionation
be required.
elements such as Pb or S.
the stable isotope ratio for S
of
hydrocarbons
separation. When using GC-ECD,
stable isotope ratios may be used to help
sources
identify
inorganic
(GC-MS)
used for the determination
The two techniques both use high-resolution
of organic pollutants
separation, or to coupling
of
the
Gas
with an electron capture detector
bonded phase GC columns
(MS) is generally
the determination
of
chlorinated
relatively
to the determination
after a chromatographic
detector used are
largely governed by the desired detection
analyses addressed in this
the
and
and the project objectives.
budgetary constraints.
chapter, mass spectrometry
columns
and with a mass spectrometer
are commonly
Mass Spectrometry
For the types of lichen
mix), followed
solvent and its polarity,
and analysis
and the chromatographic
target compounds
INAA can
of some
of elements,
and a limited
a
using silica gel or Florisil.
dependent upon the expected concentration
for
INAA is generally expensive
of reactor activation facilities
with
extraction
or an acetone:hexane
clean-up
rapid determination
a large suite
of
of
is similar to the analysis
of pigments
of extraction
The choice
the sample to remove
interferences.
pollutants
Determination
hydrocarbons.
in plants, which involves
by removal
However, lead, for example, is not routinely
determined.
Greater sensitivity and lower detection
limits can be obtained for some elements by
destructive
determine
moderately polar solvent system (e.g.,
the g g"
25-30 elements that can be routinely
compounds
pesticides
any pre- or
without
treatment.
post-irradiation
Plant samples can be
spectrometry.
nondestructively
these
have been used to
chromatography
and quantitatively
qualitatively
irradiated with thermal neutrons and the induced
radioactivity
of
Other forms
a sample is
of
retention times
from standard solutions,
and
results.
has the potential
thus does not necessarily
contribution
Confirmation is typically achieved by using a second
column with a different polarity and retention time,
or by confirmation of some portion of the samples
by GC-MS.
natural and anthropogenic
sources (Jackson and
1989, Krouse 1989). Epiphytic lichens and
Gough
sulfur species have exhibited
similar sulfur isotope ratios (Krouse 1977,
mosses and atmospheric
relatively
Miscellaneous Other Methods of Analysis
Winner et al. 1978, 1989; Case and Krouse 1980;
Taylor and Bell 1983). However, the analysis is
relatively expensive, generally not widely applicable
to nonpoint source pollution studies, and the results
can be complex and difficult to interpret.
Sulfur
digestion
(IC)
cations and anions in plant tissue.
determine
P043-
total S and IC by the combustion/IC
Other anions such as F", Br", NO3-,
are commonly
determined
samples, but their determination
complicated
quantitation.
Jackson
and
by IC in water
of
examined
washing,
in some
grinding,
parameters for the determination
from several different
in plants is
by difficulties in digesting
and titrimetric
et al. (1985)
and
of sulfur
in a variety of plants and lichens by an automated
combustion/IR method. They found that the
combustion/IR method provides rapid and precise
analyses with the analytical variance being much
smaller than the natural variability of S in lichens
Basta
(1985) determined Ca, K, Mg, and Na
in plant ash by IC, and Busman ct al. (1983)
method.
a
acid
incorporating
methods followed by
colorimetric,
analytical
and Tabatabai
determined
or combustion
turbidimetric,
detail the various aspects
has been used to
in plant material by
has been determined
wide variety of techniques
Chromatographic Methods
Ion chromatography
give unequivocal
Various
potentiometry,
the plant
volatilization losses or addition of
interferences from the digestion procedure.
without
polarography
analysis.
107
ecosystems.
electrochemical
Of
coulometry,
techniques
such as
amperometry,
have not found common
these techniques,
and
usage in plant
potentiometric
methods using ion selective electrodes
•
(ISEs) may be
only exception. Anions such as NO3-, CI", and
F" have been most widely determined using ISEs in
extracts of plant tissue. Other ISEs are available for
of
the determination
statistical evaluation,
of
estimates
the
•
ecological
of
Each
several cations and anions.
interpretation.
by the contractor
conform
analysis
results by the
to the contractor/researcher
laboratory
to the contracting
to the standards
of
where legal or regulatory
QUALITY ASSURANCE AND
QUALITY CONTROL
and
agency must
good laboratory
practice.
This requires that the following must be explicitly
defined and appropriately
•
assurance and quality
Quality
widgets, but are an important
e.g. total sulfur,
every environmental
SO42"
•
the reporting
required for a
•
the
to the
•
units, e.g. g g" Pb, wt% S
weight basis, e.g., ash weight, air-dried
at 40o C
the appropriate number of significant figures
•
the analytical
use
analytical
method limits
•
problems
encountered
of
of
QA
of
Censored
the
produced to known levels
Pb) and not
documentation
or "not detected." Questionable results
should be independently verified with the laboratory
to avoid misinterpretation
or inappropriate
conclusions in the final report. Transmittal of results
by the laboratory or the contractor in an electronic
form must be formatted so that no ambiguity exists
with respect to the requirements listed above. This
is especially true for significant figures.
In general,
reporting of more than two or three significant
figures is an egregious misrepresentation of the
data by the laboratory
are the major
studies, the
typically has
•
A QA
QA
documentation
a number
utilized
well.
or contractor/researcher,
and reviewed
database management
for field, laboratory,
and
•
QC
of
summary
agency as
agency,
and taxonomic
control
agency at a
or external verification
of
data,
vouchers.
is defined as the operations
method is in statistical
probability
statistics and
trends,
and outliers
108
control
and
A QC
laboratory
within known
and precision for a
levels of accuracy
given time period.
laboratory,
identification of data distributions,
by the contracting
samples by the contracting
laboratory,
Quality
data
generation
of
analysis,
collection,
of field,
by the contracting
associated data to show that a field or laboratory
steps:
data entry, and
and QC data
• review and validation
studies
to the
but should be
This may include submittal of blind standards
different
lichens or plants
of common
lacking or
is entirely
program should not be limited
laboratory
reanalysis
database construction,
QA
procedures.
data
of
the
inadequate, particularly with respect to
details and descriptions of chemical analysis
DATA ANALYSIS
analysis
of
components
completely
or the contractor.
The task of data analysis for environmental
are
Planning and
quality.
program.
to the laboratory
using the chemical
of
to
to specific
according
This includes planning and documentation
of the study design, the field and laboratory methods,
In many environmental
and data analysis procedures.
as "zero"
analytical
QA
control
quality
standards and data to show that measurements
values (those results determined as
be reported as
value (e.g., g g"
need a good
results.
and is a system that uses documentation
activities,
limit of detection) should
"less than a specified
proportional
or the intended
even those ecological
Quality assurance incorporates
detection
are
of
of
a project
as "pure research"
program to validate
during analysis
part
is commonly
program
However,
the data.
mandatory
show that work is performed
below
(QA/QC)
The level of effort
study.
legal ramifications
studies perceived
method used
•
control
no longer just associated with the production
documented:
determined,
the analyte
water extractable
and should be
these steps is important
issues are involved.
Reporting of Chemical Analysis Results
The reporting of chemical
significance,
levels
well documented, especially
analyzing
of
tests
confidence
program
analysis,
aspects, such as duplicate
sampling
second collector,
of
analysis
incorporates
field
and data analysis
in
replicate
the
field by
samples and
standard reference materials,
and spot checking
data entry and data analysis
results.
of
a
aspects of
Various
QA
design, field methods, laboratory
have been discussed
by Taylor (1987) and Dux (1986). In a laboratory
QA program, documentation of the daily use and
analysis of external or internal calibration
quality control
aspect is the analysis
problematic
NIST
reference materials in
A
botanical
U.S.
is the primary
of an adequate QA program.
In addition to an understandable, concise
description of a project's scientific conclusions,
final report should include clear, thorough
large number of determinations
is no longer available.
et al. (1987)
Gladney
number of other biological, geological,
of:
descriptions
and
The main botanical SRMs
NIST 1515 Apple Leaves, 1547
and study design
•
study objectives
•
field and laboratory
methods
environmental
SRMs.
•
data analysis procedures
now available
are
•
QA
Peach Leaves,
1573
Leaves,
Tomato
1575
Horwitz
References
provides
SRMs.
endless wild-goose-chase
certified,
additional
in
constituents
range as lichens.
Elements
and noncertified
constituents
methods are acceptable
these standards do not always
sufficient
references.
same concentration
the
•
such as S are not
values for various
requirements
reference materials and their suppliers
Without
The cost
of SRMs,
matrix matching,
has
well
frequently
of good
measurements
to the data reporting
outlined
above,
a report's
independent
conclusions
a study without
or future
undetectable
biases
cannot be made.
served as an excuse
inclusion and analysis of SRMs in
studies. For any study currently
involving trace element determinations in lichens,
inclusion of one or more SRMs in the QA program
to avoid
of obscure and incomplete
such documentation,
of
repetition of
validation
as the lack
detail and does not lead one on an
of sample duplicates and SRMs
have been
(1989a,b).
as
of
all QC data pertinent to verifying the
quality of the raw data, such as the analysis
•
by Roelandts
descriptions
the original source
all raw field and laboratory
This latter problem is especially true for organic
pollutants, for which no botanical reference material
exists. Lists of additional international biological
compiled
if
The report should include addenda with:
conforming
have not been determined.
published
to previously
and Albert (1991) review the variability associated
with concentration estimates for several of these
have certified
used
and software
program plan
Pine
Needles, and 2695 Fluoride in Vegetation.
Unfortunately,
a
have
results for these two SRMs and a
analytical
compiled
the absence
constituents
and noncertified
certified
a regular
This is especially true where a study
be
may
repeated in the future by the same or another
contractor.
Minor changes in analytical methods due
to incomplete documentation may artificially
introduce biases or trends in the data, especially in
for
in NBS 1571
Orchard Leaves has been published for many
However, this SRM,
different analytical techniques.
1572 Citrus Leaves,
its
NBS
with
replacement
along
the
of
methods, details, or raw data typical
One
supplier of certified
This should
and lack of
journal article.
of
standard reference materials.
methods, data
interpretation.
not be done with the same brevity
blanks,
and standard reference materials is mandatory.
particularly
analysis, and ecological
standards,
samples, sample duplicates,
address the issues of study
report should thoroughly
programs directed at
laboratories
chemistry
analytical
the
environmental
CASE STUDIES ON THE USE OF
CHEMICAL ANALYSIS OF LICHENS
Taylor (1985) reviews
numerous aspects of the use of SRMs in a QA
should be mandatory.
The use of lichens and bryophytes as biomonitors
of chemical contaminants, including metals, sulfur,
program.
fluoride,
radionuclides,
and organic
pollutants,
has
been reviewed by Burton (1986)(see also Wilson,
Other reviews which focus on lichens as
1991).
FINAL CONTRACT REPORTS
indicators
of metal pollution have been done by
Final contract reports for environmental studies
using chemical analysis of lichens for biomonitoring
James (1973), Nieboer et al. (1977), Nieboer and
should conform
Anderson
to guidelines
Richardson
set by the agency
requesting the report; for example,
(1981), Martin and Coughtrey
and Treshow
(1984), and Puckett
(1982),
(1988).
Most studies involving chemical analysis of
see Harbour
(1987). These guidelines primarily concern style and
format issues, not scientific content. A final contract
lichens have focused on determining
baselines or examining
109
elemental
spatial or temporal
trends
Level
2, 7.2
South Unit
North Unit
Level L, 72 km
km
Level 3, 0.72 km
Level 4, 0.10 km
Level 5, Lab
Replicate
n
n
5- Level,
Unbalanced, Nested Analysis of Variance Design
Figure 2. — Schematic of unbalanced, hierarchical ANOVA sampling design
(data from Gough et al. 1988b).
with respect to pollution sources. Chemical baselines
have been defined in various ways. Usually a
baseline refers to a specific set of conditions and
point in time (i.e. when the samples were collected)
and not to historical or pre-industrial conditions.
One definition of a baseline is the expected 95%
range, which is the mean plus or minus two standard
deviations.
For lognormally distributed data (Tidball
and Ebens
typically
the case when the
of
analytical
methods and/or there was very little natural
between samples, as in the case
with respect
When
range.
range for those
elements in which the laboratory
that the variance
et al. (1988a,b)
Gough
chose not to express a baseline
of
sulfur
variability, it is unreasonable
to natural
50% or more
of
error is large
the laboratory
to calculate a baseline
(GM/GD2) to (GM X GD2)
the
detection for the
variability
concentration.
1976), the expected 95% range is:
in
mean concentration
lichen was very near the limit
error contributed
the total variance.
They also found
attributed to within-site variability
deviation,
(i.e. replicate samples at 10-100m) combined with the
laboratory variance generally represented 70-100% of
the total variance (fig. 4). As these studies point
the latter
out, regional differences
where
GM
and GD are the geometric
mean and
Gough et al. (1988a,b) used
respectively.
definition to determine baseline ranges for
a number of elements in Parmelia sulcata at
Theodore Roosevelt National Park, North Dakota,
and in Hypogymnia enteromorpha and Usnea spp. at
Redwood National Park, California. In both studies,
spatial variability was estimated using an unbalanced
nested ANOVA design. For example, in the
Theodore Roosevelt National Park study, differences
in elemental content in P. sulcata between park units
(i.e. about 70 km apart) and within a park unit were
examined with respect to the analytical measurement
error (fig. 2). As shown in figure 3, for some
elements a large proportion
attributed to laboratory
of
the total variance
measurement error.
in element concentrations
must be large with respect to the variability
attributed to within-site
differences
and laboratory
error to be detectable.
In numerous studies, elemental concentrations
lichens have been clear indicators
However,
lichens will probably
quantitative
estimates
accompanying
of
Empirical correlations
in
pollution.
not give very reliable
deposition
conventional
of
without
atmospheric
monitoring.
between heavy metal levels in
lichens and mosses and actual measured deposition
from bulk precipitation
collectors
have been observed
in several studies (see review by Puckett, 1988). In
monitoring, the use of
the absence of conventional
was
This was
110
Geometric mean, ppr
ppm
100
51-
i
5
2
II
i
8
5
5
>
Hypogymnla tJSJS Parmella
Usnea
Figure 3. — Example of percentage of total variance attributable to laboratory error for the
and Parmelia sulcata
analysis of Usnea spp., Hypogymnia enteromorpha,
(data from Gough et al. 1988a,b).
inferred trapping
efficiencies
to provide
potential
information.
monitoring
of
the percentage
The trapping efficiency
a given
and/or
To estimate
suburban areas.
foliose
the
trapping efficiency for an element, the total time of
from radionuclide
biomonitors
served as
of point and nonpoint source pollutants
for metals, sulfur, and fluoride,
measurements
sites in the
lichen.
Whereas lichens have frequently
(i.e. the lichen age) is needed. The age
of the thallus may be estimated from known growth
or possibly
Their
They also found that the lead was
apparently retained at ion-exchange
accumulation
rates
lead fallout rates.
estimates agreed with levels expected for rural and
is
surface area that are
retained by the lichen.
actually
estimate retrospective
quantitative
metal ions in precipitation
falling on
particulates
from Great Falls, Maryland, and to
baltimoremis,
by lichens has the
retrospective
they have only
recently received attention as suitable monitors
of
such as l37Cs or 210Pb/210Po ratios.
organic pollutants.
Carlberg
Pb is a decay product of naturally
occurring radon which diffuses into the atmosphere
from the soil, and its deposition rate is known for
al. (1984), Thomas
(1986), Villeneuve and coworkers
Because
specific
210
geographic
regions, the maximum
(1984, 1988), and Bacci and coworkers
1986,
theoretical
thickness
of foliose
concentration
area.
its in-situ
of
et al. (1987,
atmospherically
daughter
efficiencies in
Antarctic
for chlorinated
hydrocarbons
and its metabolites,
PCBs, lindane and other
1991) used
210
derived
Pb and
Po to infer trapping
lichen, Flavoparmelia
a saxicolous
111
the
et al.
and other
These studies indicate that
of
per unit of surface
(Bacci
et
1991) have analyzed lichens
Spitsbergen, Sweden, France, and the
organic pollutants.
least ng g"
because the
Thomas
et al.
isomers, and
hexachlorocyclohexane
hexachlorobenzene contaminate lichen
lichens varies with age,
must be determined
Schwartzman
measurements
However,
Focardi
from Norway,
of
Pb in the lichen can be
calculated.
As a first approximation, the inferred
trapping efficiency for lead is the ratio of the
measured
Pb concentration in the thallus to the
Using measurements of
theoretical concentration.
lead in thallus of varying ages, retrospective fallout
concentration
rates can be estimated.
et al. (1983),
DDT
tissue in at
levels even in the remote environments
Arctic and Antarctic.
10 or 100m varlanc*
+ lab
variance
•too
o
©
V
n
3
■o
8
5
1
1
«*-"
r~~ni Itenea
I
-•■*».
Hypogymnla
-^
^S Parmella
Figure 4. — Example of percentage of total variance attributable to spatial variance
(closest spatial stratification level) and laboratory measurement variance
(data from Gough et al. 1988a, b).
ACKNOWLEDGEMENTS
Biological Monitoring of
(Plants). Monitoring
and Assessment Research Centre, King's College
Burton, M. A. S., 1986.
Environmental
The authors thank Fred Rhoades
and Margaret
Wilson for their assistance in preparing the outline
for this chapter at the NPS/FWS Lichen Workshop,
April 9-11, 1991.
The use of trade names is for descriptive purposes
only and does not imply endorsement by any Federal
London,
University
MARC
London.
Technical
Busman, L. M., R. P. Dick, and M. A. Tabatabai,
1983.
Determination
plant materials.
of
in
total sulfur and chlorine
Soil Science Society of America
Journal 47:1167-1170.
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Van Nostrand
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Hypogymnia
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metal fallout pattern from lichen and soil assays.
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handling, and analyzing
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Lessons from
Seven Case Studies
Ken Stolte, Robert Doty, Kathy Tonnessen, Rich Fisher
This chapter provides
information
a conceptual
model for integrating
in the earlier chapters
provided
of
this report.
the
The model is
used to help analyze seven possible air resource management scenarios.
Each scenario is illustrated with
a case example
of an actual air pollution
study using lichens.
Lichen monitoring
intended to directly
support air resources
can be designed and conducted in many ways,
management activities
on the specific air quality issue of interest. For example, the
of plots and the frequency of sampling will differ depending on
whether the air pollution stress is from a particular point source or a large
depending
placement
Despite such differences,
urban area.
designed to follow a conceptual
most lichen monitoring
can be
model.
in this chapter can be used to assist in deciding
The information
whether or not lichens can be used as bioindicators
or bioaccumulators
of
air pollution.
THE CONCEPTUAL MODEL
pollution
and physiology can be
This work is more time consuming and
on lichen chemistry
performed.
The information presented in other chapters of this
report can be interrelated as shown in figure 1, the
Model.
Conceptual
modeling
monitoring
predicted and actual pollutant
This, together with
the problem
a study design.
and 7 for details.
A
to characterize
loadings,
monitoring
of
transplants
active
(Chapter
6)
The identification of indicator
species is only needed when no previous
is used
Chapters 2,4,5,6
characterization
by studying
becomes necessary.
respectively.
description,
See
by air pollution to the
extent that species have been extirpated,
The model uses air quality
and aerometric
to formulate
expensive.
For areas already influenced
information
exists about the native species and their response to
the air pollutant
the site
3) is essential for establishing a foundation
for specific lichen analyses. A community analysis
facilities
(Chapter
of interest.
and well-designed
Elaborate
fumigation
exposure-response
studies
for this work.
are necessary
(Chapter 4) is used to identify the grouping of lichen
species and other environmental variables that define
the terrestrial
ecosystem.
A floristics
sensitive
POLLUTANTS OF CONCERN
study (Chapter
2) determines the presence or absence of species.
many instances, this information is sufficient to
In
The following list includes
can be obtained
air pollutants which
could affect lichens.
5) and to assess
the air pollution effects on lichens.
This information
identify
species (Chapter
from a lichen expert.
Primary
When the above steps do not yield conclusive
results or when no previous information exists about
the native species or their responses to air pollution,
elemental analyses (Chapter 7) of the effects of air
116
of sulfur
•
Oxides
•
Hydrogen
•
Oxides
of nitrogen
Meteorology
Emliilont
Topography
|
air quality
Problam
mod»Hng
Description
L
Domrnunlty |
Analysis
Figure 1.—Conceptual model for lichen-based air pollution monitoring.
Numbers near the boxes refer to pertinent chapters in this report.
fluoride
PM10
Toxics (metals, organics,
from the point source.
Climate
operate principally
heights can result in the fumigation
Hydrogen
(especially
sporadically,
radionucleides)
temperature and
humidity)
Topography
during periods of high demand.
and geography also affect the level of
the
At night and often in winter,
plumes can be channeled along valleys, allowing
Sulfates (wet or dry)
(wet or dry)
pollutants to accumulate in low spots. Vegetation
canopies can absorb plumes, altering the exposure of
lichens located close to the ground.
Ozone
Ammonia
Wind direction is
meteorological
POLLUTANT EXPOSURE FACTORS
source characteristics
concentration
include
most important
affecting
point-source
while temperature, humidity
sunlight contribute
Due to meteorological
tall stacks and high effective
the
variable
pollutant exposure for a given lichen community.
Mixing height and wind speed help determine plume
rates, operating schedules,
and effective plume heights.
conditions,
that intersect
lichens experience.
Nitrates
pollutant
Plumes
elevated terrain increase the amount of pollutant
Secondary
Important
power plants, which
such as "peaking"
lichen exposure to pollutants.
pollutant types, emission
of vegetation far
Some point sources operate
pollutants.
plume
117
to the chemical
and
transformation
of
CASE EXAMPLES
SEVEN AIR RESOURCE
MANAGEMENT SCENARIOS
The case examples describe the use
Most air pollution situations fall into one of
following seven categories:
A) An existing point
expected
Since each past and future situation
that is not
necessarily
source
solutions
the preferred
of
lichens as
each scenario.
is unique, these
they are not
to the scenarios.
lichen biomonitoring projects, some,
When designing
all, or none
that is expected
of
in past studies illustrating
cases are presented only as examples;
emissions.
to change
B) An existing point
to reduce
source
bioindicators
the
the information
from these examples
may be useful along with details from the rest of this
document. The case studies are brief summaries;
emissions.
C) An existing point source that is expected
to increase emissions.
refer to the original
D) A new point source (e.g. through the
Prevention of Significant Deterioration
references for additional
details.
SCENARIO A
Process).
E)
A
Constant Point Source Emissions
region affected by air pollution with
increasing regional loadings.
A
F) A region affected by air pollution with
declining regional loadings.
G)
vitality
region where the loadings are known or
thought to be so low that no adverse
impact to resources has occurred.
monitoring
case example
illustrates
source.
A
X
X
X
B
X
X
X
D
ix
X
of
a phosphorus
(1980) investigated the
plant in Newfoundland,
X
X
E
ix
F
X
X
G
X
X
X
Sidhu and Roberts
7
vegetation damage.
X
of
X
X
X
X
pollutants
1969.
emitted were
(HF) and silicon tetrafluoride
X
the lichen
(1976) described the nature of the
They evaluated the composition
communities
in four damage zones
around the plant and at 12 similar control
X
sites
Two of the most common
species, the terrestrial lichens Cladina rangiferina
(L.) Harm, and Cladina stellaris (Opiz.) Brodo., were
analyzed for fluoride concentrations in the tissues.
The morphological condition of the lichens was
examined in each zone.
The concentration of fluoride in the samples was
found to be inversely correlated with distance from
distant from the plant.
X
X
X
6
X
X
and epiphytic
northeast of the plant due to the prevailing winds.
6
X
X
terrestrial
This facility had been operating since
By late 1974, damaged vascular vegetation
(SiF4).
around the plant covered 1 1 ,400 hectares, mostly
CHAPTER
A
pollutant
The greatest impact from a single source
hydrogen fluoride
figure 1 and pertinent chapters in this report.
(1) is Study Design and not a chapter.
4
gradient of
condition
decreases with distance from the
The primary phytotoxic
air pollution monitoring summarized
in relation to numbered model components of
(D
A
Roberts and Thompson
effects
1. — Scenarios of
c
and morphological
Emissions
lichens.
SCENARIO
from the source.
Canada, on the surrounding
3
in the
number or
Case Study of Constant Point Source
A
shown in table 1.
2
the
would be expected near that source.
summary
of the model components (the other chapters in this
volume) illustrated in each of the seven scenarios is
Table
in this chapter
in
might be expected as the ground-level
concentration
and research and
each scenario.
air pollution may
sensitive genotypes and species due to
species composition
stressor might have on the lichen
methods that might be appropriate.
loss or reduction
distance downwind
of these scenarios is briefly discussed below
in terms of the stressors (e.g., pollutants) of concern,
species and/or communities,
of
of
lichen populations
chronic or acute exposure might be expected at some
Each
the
A
vicinity.
A
the effect
single existing point source
already be affecting
X
X
118
in the direction of the prevailing southwest
Fluoride concentrations ranged from a peak
to remove
the plant
could result from improved
winds.
pollutants from the stack emissions, a switch to
cleaner fuels, or a reduction in activity of the plant.
of 2830 ppm (dry weight) in Zone 1 closest to the
source to an average of 6.4 ppm in the control areas
outside of Zone 4, about 9 km from the source.
Fluoride concentrations in Zone 1 ranged from
525-2830
emissions
downwind of the plant should be
in condition
under these circumstances.
To measure improvement
in lichen condition, we could measure the reduction
in toxic element concentrations, improvement in
morphological condition of existing lichens, and
establishment of new or returning sensitive species.
communities
expected to show some improvement
between the morphological appearance of the lichens
and the accumulation
individuals of
the
of fluoride in
indicator
All
the tissues.
species found in Zone
1
were amorphous
(without structure) and discolored,
while about 25% of the individuals of the indicator
species in Zone 4 were only slightly
some structural
degradation.
discolored
with
Case Studies of Declining Emissions Around
Point Source
Fluoride concentrations
in C. rangiferina ranged from 6.4 - 1338 ppm along
the gradient from Zone 1 to the Control Zone.
The
researchers observed a similar pattern of fluoride
accumulation in the 1974 needles of balsam fir (6.1 -
a
Showman (1975a) mapped the distribution of
lichens in 1973 at 128 sites around the isolated
Muskingum River power plant in
He found that the lichen
Flavoparmelia caperata was a particularly sensitive
species and was absent from many sites near the
power plant. Air quality monitoring in subsequent
years showed a reduction in SO2 emissions as a
result of design changes to the power plant in 1972.
coal-fired
237.0 ppm), soil humus (2.1 - 24.2 ppm), terrestrial
bryophytes (11.3 - 2255.0 ppm), and air (0.1 - 5.2 ug
southeastern Ohio.
F/m*).
Comparison with Conceptual Model
A) Pollutant Exposure:
Ambient Fluoride (ug
F/m ); meteorological variables
Researchers
caperata
had recolonized
a
few sites, evidenced by
at tree
with this species and there was no longer a
recognizable void of this species near the power
plant. A community-type
analysis showed similar
results: the number of corticolous species at sites
within the 1973 void zone increased from 2.0 in
1973 to 3.8 in 1976 to 4.7 in 1980 (Showman,
1981). In this study the distribution of sensitive
4 damage
lichen species and overall
lichen community
were useful measures to delineate areas
of
structure
poor air
improvement in air
in
reductions
stack
emissions.
following
quality
quality in 1973 and to monitor
No
Comparison with Conceptual Model
A) Pollutant Exposure:
No
SCENARIO
sites around the
bases. By 1980, many more sites were recolonized
total fluoride in the
F) Chemistry/Physiology:
two lichen indicator species and other
epiphytic lichen species.
G) Transplants:
128
by 1976, Flavoparmelia
small pieces of thalli in sheltered bole fissures
Cladina rangiferina
D) Indicator Species:
and Cladina stellaris
E) Community Analysis:
reevaluated the
power plant annually;
B) Site Characterization: wind speed and
direction; temperature and rainfall
patterns; vegetation communities; tree
species associated with forest types; other
vegetation communities; and parent
material and soil types.
Control sites
comparable to damaged sites.
C) Floristics: lichen floras from
zones; 1 control site
consequences of reduced stack
should be favorable, and lichen species and
The ecological
There was a strong correlation
ppm.
technology
around
B) Site Characterization:
selected; elevation
B
SO2 monitoring
power plant
Unshaded trees
C) Floristics: Initial flora to select sensitive
species
Decreasing Point Source Emissions
D) Indicator Species:
Lichens have been used to indicate improvement
in air quality as emissions of toxic elements from a
point source decline. This reduction in emissions
SO2 sensitive
119
Parmelia caperata L. -
1981-1982, and that Zn and Cu were higher in
E) Community Analysis: Quantified
community analysis
No,
recolonization
natural
SCENARIO
An increase in emissions
sensitive genotypes
increase.
of
decreased number
only
species, increased mortality
of
B) Site Characterization: little information;
host tree, number of replicates, and
and Tel Aviv, Israel.
air pollution.
in heavily
use
of
Some
C) Floristics: no community analysis; only
indicator species identification
MW
Sharonim,
This
active monitoring
D) Indicator Species: one common species
{Ramalina duriaei (De Not.) Bagl.)
selected for transplants
of
sites were
the biomonitoring
areas near Tel Aviv and other
urbanized
small cities; one was a control
a 1400
of
began operation
in
1981.
of their
sizes of the
distance
from affected areas, the small
urban
of
purpose
These were
regarded as point sources because
areas, and the nature
E) Community Analysis: not done (not
site. The Maor David,
power station located near
coal -burning
1982, Fuchs
July 1982.
absorption
1983), and then exposed a
the same species from July 1981 -
and Garty
second group
of
He used X-ray fluorescence
spectrometry
concentrations
of
Pb in the lichens
and atomic
SCENARIO
to compare the
among the sites and between the
New Point Source Emissions
pollutants
near the urban centers and the power plant than in
from relatively
between
indicated
lichens
D
the elements Ni, Cr, Cu, Zn, and
The thalli of R. duriaei nearest the urban centers
in 1981-1982 were chlorotic after one year of
Metal concentrations were higher at sites
exposure.
of
chemical analysis of
data.
two time periods.
tissues
the study)
G) Transplants: This method illustrates the
effective use of transplants and the value
of corroborative pollutant accumulation
twigs from
1979 to March 1980 (Garty and Fuchs
February
of
F) Chemistry /Physiology:
toxic metals
their wind plumes.
Garty exposed lichens on transplanted
on the host tree
transplants
were given
Garty (1987) transplanted the common epiphytic
species Ramalina duriaei (De Not.) Bagl., to 22 sites
effective
of
placement
Case Study of Increasing Point Source
Emissions
study illustrates
the coal burned at the
in orchards
of
the
affected area around the point source.
located near Sharonim
of
A) Pollutant Exposure: metal content of fuels;
amount of vehicular traffic; Zn spraying
We can also
less tolerant individuals, and expansion
vehicles
Comparison with Conceptual Model
will be
deterioration,
expect to observe morphological
of
plant during that period.
from the
exposure to pollutants,
present after the emissions
They attributed Pb
1979-1980.
were linked to the content
If
lichens.
species and individuals
the more tolerant
Pb/Ni,
the associations
which still used leaded gasoline and an increase in
miles driven during 1981-1982.
High Cr and Ni
concentrations found in lichen tissues in 1981-1982
from existing point
are eliminated
analyses among metal
patterns to a 16% increase in the number
affect existing
by chronic
population
indicated
orchards during
C
Increasing Point Source Emissions
sources can negatively
Correlation
For
Pb/Cr, Pb/Mn, and Zn/Mn to be significant.
example, the Pb/Zn ratio increased from 0.19 in
1979-1980 to 2.11 in 1981-1982.
Researchers
attributed the reduction in Zn concentrations to
reduced spraying of Zn in the citrus and pecan
No
F) Chemistry/Physiology:
G) Transplants:
occurred
1970-1980.
concentrations
from
1970-1980
the control
site.
Lichens can
be used to map the distribution
emitted from a newly
unit in a previously
undisturbed
good air quality
installed
of
A
change
conditions
to an
area.
air
industrial
in ambient air to phytotoxic
can be observed in lichen communities
increase in pollutants
Comparisons
concentrations
and 1981-1982 study periods
that Pb, Ni, and Cr were higher in
through increased mortality,
120
decreased morphological
of
condition
survivors,
elevated element
the stack.
The severity
of
effects usually
with increasing distance from
the
to the air pollutants,
sensitivity
in tissues, decreased cover of sensitive
species, and a general decline in species diversity.
Under conditions of low moisture the magnitude of
the impact will be lessened.
The most severe effects
usually occur within 1-5 kilometers of the source,
depending on the size of the source and the height of
concentrations
Parmelia
included:
Usnea spp., Ramalina spp., Parmelia
perlata,
Parmelia glabratula, Parmelia caperata,
Parmelia sulcata, Evernia prunastri, Hypogymina
The first two
physodes, and Lecanora conizaeoides.
subrudecta,
species were particularly
sensitive,
and their
with the 40
'contours'
pollution
ug/m3 SO2 (annual average)
recorded by SO2 monitors in the area. Only
Lecanora conizaeoides seemed relatively unaffected
by the emissions and was still present at all sampling
distribution
declines
source.
Case Study of New Point Source Emissions
Lichen species were evaluated on lands
the Esso oil refinery at Fawley,
Hampshire, England (Morgan-Huws and Haynes
A small refinery was built in 1921; a larger
1973).
unit was added in 1952. Sulfur dioxide and other air
pollutants were emitted from the furnace stacks from
30-50 meters in height into the surrounding low-lying
Little
landscape of the Southampton Water area.
coincided
approximately
sites, even those close to the emission
source.
surrounding
Comparison with Conceptual Model
A) Pollutant Exposure: information on
addition of SO2 sources; isopleths of
around
B) Site Characterization: general landscape
description; collection methods detailed
smoke was emitted due to the high combustion
The areas surrounding
temperatures.
the
refinery
C) Floristics: detailed
were a mixed agricultural-heath-forest-residential
No pre-1921 lichen records existed, but
around the Fawley refinery in
1921 is presumed to have been similar to floras
found in nearby areas of similar vegetation and
habitats but without air pollutants.
The town of
Fawley is at low elevation on the Southampton
maritime air-masses,
95-100%
common.
the area is influenced
with relative humidities
E) Community Analysis: no abundance
cover of species recorded
by
F) Chemistry/Physiology:
from
There was some background
G) Transplants:
of SO2 and smoke from the ribbon-like
urban development in the area.
Lichens were sampled from the trunk of a
common tree in the area (Quercus robur) for
comparison of sulfur dioxide effects. They examined
influence
the southwest-facing
half-sector
of
the
Remote clean air areas
becoming
Trees selected were well-established
(open-grown).
(dbh 0.5
and free-standing
Many sites included only
transport
a few trees
years
of
of
increasing
important to
from continually
Small communities
growing large
and rural areas in
growing and adding to the
loading in remote areas. It is
establish floristic and community
soon as possible for trend comparison
The transport
of
secondary
45
resulted in pollutant
relatively
There was a
loadings
such as
pollutants,
sulfates, nitrates, ozone, and particulate
exposure to the small source and 14 years
site.
pollutants
with future studies and analyses.
metals, has
in areas of the country
remote from large point sources
of
pollution such as power plants, refineries, or
smelters. To determine the impact of these regional
marked decrease in most lichen species around the
emission
the western US are
pollutant
baselines as
was more dense in areas where ambient
exposure to the larger source.
of
of
the West are also
carefully devised to obtain statistical correlations
relative to the pollution gradient.
As a result, site
pollution exposure gradients changed abruptly.
Lichen floras were evaluated in 1966, after
E
dirty from the long range
increasingly
urban areas.
each ( 4 trees), with the general approach patterned
after Skye (1968).
The sampling design was
replication
No
Increasing Regional Air Emissions
the trunks to a
This part of the trunk
entire macrolichen flora for the area.
meters), erect, undamaged,
or
No
SCENARIO
height of 2 m above ground.
included
around
D) Indicator Species: relative sensitivity of
species inferred from field gradient
analysis
vegetation
Water, and consequently
flora centered
one host species
landscape.
the lichen
SO2
sources
Species present at the site, in order of
emission
121
sources, lichens may be used as
bioaccumulators
concentrations
of certain
of pollutants
Historical
elements.
C) Floristics: Only a single "indicator"
species, Flavoparmelia baltimorensis, was
used as a bioaccumulator of lead and
sulfur.
such as lead or sulfur
can be measured in herbarium
tissue samples of
lichens from the same general area as the
This is a situation in which
specimen.
information is collected to determine the
contemporary
"baseline"
rate
of
of
degradation
increasing
inputs
of
regional
D) Indicator Species: Researchers selected an
indicator species that was common to the
park to ensure that repeat surveys would
yield specimens for chemical analyses.
in response in
an ecosystem
pollutants
to wilderness
areas.
E) Community Analysis: No, not part of the
One would not expect
study plan.
detectable change to communities in
Case Study of Increasing Regional Emissions
In the Mid-Appalachian Mountains of western
Virginia, Shenandoah National Park was created in
1936 to preserve second-growth
Due to the proximity
ecosystems.
area to planned and projected
both point and non-point
the differences
concentrations
in
the
of
sources
F) Chemistry/Physiology:
this class 1
of
emissions,
commissioned
analyzed
tissue
a study
use
between contemporary
of
herbarium
SCENARIO
Lawrey and Hale (1988) located eleven sites
within the park that had been sampled during the
period of 1933-58. Tissue samples of the lichen
Flavoparmelia baltimorensis found in the herbarium
collection at the Smithsonian Institution were large
enough to allow elemental analysis for lead and
using inductively
coupled
States, from the Ohio River
coast,
Atlantic
is heavily populated and
valley
includes much of the nation's industrial development,
both of which contribute to air pollution in this
region. While the population continues to grow,
to the
plasma techniques.
presumably
traditional
in regional emissions
are being
replaced by
forms of development.
Furthermore,
EPA
sections of the
states
and the
Clean
Air Act Amendments of 1990 that call for the
of the total amount of sulfur dioxide and
are
implementing
reduction
nitrogen oxide emissions.
Lichen analyses can be
of
used to measure the progress
activities.
Documentation
general health
of
of
the
these control
flora and
lichen communities
the
is needed to
establish a baseline so we can detect vegetation
improvements
Comparison with Conceptual Model
Elemental analysis
lichen samples was used as a surrogate
for instrumental monitoring. Gaseous
pollutant and precipitation monitoring
data are also available for this park.
industries
less-polluting
sulfur.
A) Pollutant Exposure:
F
The eastern United
as the result of
of lead linked to
changes in the burning of leaded gasoline in vehicles.
However, sulfur concentrations in lichen tissues
increased significantly at all but three of the sites,
with those at high elevations (over 1000 m in
elevation) showing the largest percentage change in
sulfur (ug/g dry weight). This points to
long-distance transport of sulfate as the source of the
additional
the
Decreasing Regional Emissions
decreased at all sites during the
time between collections,
the change
of
specimens.
No
G) Transplants:
1988).
sulfur,
for lead and
Physiological
were not noted because
changes
lead and sulfur
specimens
Lead concentrations
Researchers
samples
sulfur concentrations.
Park
(1984-86) and
(1933-58) of the lichen
Flavoparmelia baltimorensis (Lawrey and Hale
historical
air" areas.
"clean
forest
sources, the National
Air Quality Division
Service
of
hardwood
as air emissions
are reduced.
Case Study of Decreasing Regional Emissions
Recolonization by epiphytic lichen species around
the northwest areas of London, England was
of
determined by evaluating
species at 29 sites in 1980
(annual SO2 averages around 130 ug/m3), and
re-evaluating the same sites in 1988 (annual SO2
B) Site Characterization: No information on
site characteristics; the same sites were
visited at different times to make the
comparison.
averages 29-55 ug/m ) (Hawksworth and McManus
1989). This area includes parks, reserves, gardens,
cemeteries, woods,
additional
122
An
in 1988 to more
and other natural areas.
22 sites were added
document
thoroughly
patterns of lichen
the spatial
The lichen flora occurring
recovery.
site was determined
lists were probably
host trees, the relative
F) Chemistry/Physiology:
not complete.
recorded the species identified,
Researchers
frequency
of
types of
occurrence
site;
G) Transplants:
no
no
on a
1-5 scale, and the diameter or length of the largest
foliose and fruticose species at each site.
The lichen floras for both years were classified
SCENARIO
to the species classes devied by
according
at each
species
flora relative to Hawksworth
and Rose qualitative scale (1970)
therefore,
1988;
of
characterized
during a .5-1.0 hour collection
period per site from April-October
the species
and size
on trees at each
G
Regional Air Pollution Emissions-Clean
Areas
Hawksworth and Rose (1970). By 1980, the lichen
flora had shown improvement in diversity and vigor,
compared to London floras established earlier (e.g.,
Laundon 1970), and it was postulated then that
further improvements in lichen flora could be
expected if the annual SO2 levels fell to 40-50 ug/m3
(Rose and Hawksworth 1981). [For reference, the
of the United States,
Basin of Colorado and the
Sublette Basin of Wyoming, have experienced sudden
and dramatic increases in development, either through
increasing industrial activity or related secondary
Because of their topographic locations and
growth.
annual average SO2 concentrations
proximity
found in class I
of
areas in the eastern U.S. are in the range
3.5-21.0
subject
ug/m3 for the period
of London
and additional
similar
were found that had not
to note that the recolonization
S02-sensitive
species occurred
rapid
The potential
effects on
in regions such as these are
those anticipated in Scenario D, where we
to
the introduction
hypothesize
been recorded by Laundon (1970). Eight species had
not been recorded in the area for over 200 years. It
is interesting
of
concern about the effects
lichen populations
that had not been observed in 1980;
1 1 species
areas, these basins are the
to wilderness
of
increases in air pollution.
1989-91.]
The authors observed 26 species in the northwest
area
isolated regions
Recently,
such as the Piceance
previously
clean area.
expected area ultimately
of
energy development
of
a new source
In this case, however,
to a
the
affected by this regional
will
be much larger than in
Scenario D.
at a faster rate than
would have been expected from the Hawksworth and
Rose species classes. This was attributed to the
rapid improvement in air quality from 1980 to 1988.
This study illustrates that extremely high ambient
Ryan (1990) surveyed lichens in a number of
wilderness areas in California. The Marble Mountain
SO2 levels exert a pronounced
wilderness
occurrence
of
lichen
microenvironmental
species.
conditions
influence
probably
on the
of temperature and
are less
did not change dramatically
of
within
conditions
The largest point source
California is thought to be
of
a
is a pulp mill
boundary.
the city
Comparison with Conceptual Model
in the last 20
A) Pollutant Exposure:
report,
Comparison with Conceptual Model
A) Pollutant Exposure: characterized
mean winter SO2
emissions
located over 100 km from the wilderness
a
years.
B) Site Characterization:
in Northern
clean area, with few air pollution sources upwind.
Changes in
moisture due to urban development
factor, since environmental
Case Study of Clean Air Areas
of the
of emissions
this is not part
but a state inventory
does exist; no sources
are identified in
this inventory.
ambient
B) Site Characterization: a detailed mapping
of the plant communities or ecotones was
not performed, but the major land forms
of the wilderness have been identified
and the wilderness Air Quality Related
Values (AQRVs) were listed for this
This list included lichen species
region.
among the potentially sensitive AQRVs.
host trees; time at
each site
C) Floristics: intensive floristic characterization
D) Indicator Species: characterized species by
sensitivity to SO2
E) Community Analysis: relative abundance
C) Floristics:
123
Most of Ryan's work was
compilation
of
an inventory
of lichen
geologic
species; there was no existing information
on the lichen flora of this wilderness area.
of
of sulfur
Lichen surveys in wilderness areas often result in
of new species or morphological
variations of previously-named
species. In recent
years, floristic surveys and other lichen inventory
efforts have also resulted in the identification of new
species. In addition to the lack of good keys or
guides to lichens in various areas, such discoveries
result in difficulties in identifying lichens in the field
for novice lichenologists or others who may have
only a general background in botany.
the discovery
were
No
Lack of Dose-Response Information
Most information
on species sensitivities
in the literature is based on the exposure
Using lichens to monitor air quality is not often
and usually
straightforward
lichens to sulfur dioxide.
requires considerable
Here are a few potential problems
should be kept in mind when conducting
quality
on the susceptibility
that
Nash
lichen air
rigorously
for
lichen communities,
of
it may be difficult
pollutants.
in future studies.
to be combined
exposure.
Natural Variability in Characteristics
The use of the morphological status of lichens as
an indicator of pollution stress is problematic.
The
natural variability in thallus geometry and chemistry
should be measured in both baseline and treatment
information
significant
to air pollutants
designed
air
has been inferred
fumigation studies.
from other sources
of
monitoring
results are suggestive but not
or 5% levels. For example,
of a single lichen species
may suggest a pollution-related
significant.
causes may be possible
concentrations
of
changes in the physiology
Anthropogenic vs. Natural Sources of
Elements
When chemical analysis of lichen tissue is
or drought.
community
cause, but may not
Other, non-pollution
explanations,
such as aging
information on changes in
floristics coupled with known increased
However,
pollution loadings in the area can help support
weak inference
the
due to (1) contamination
and analysis, (2) natural
and
subspecies
into the same effort.
at 1%
be statistically
it may be difficult to distinguish
of
may be necessary to make decisions
when statistical
areas.
background
and
To address these gaps in knowledge, monitoring
pollution and research on dose responses may need
It is also
of
the spatial distribution
Corroboration
handling,
toxics,
Much of what has been asserted about
lichen sensitivities
must be done very carefully
in a given area relative to pollutant
relative contributions
ambient
to known concentrations
from a few statistically
collection,
fumigation experiments
between the genetic differences
essential to monitoring,
performed,
(Sigal and
have never been
There is also little information on the relationships
that support
their susceptibilities
lichens
to ozone
gaseous air pollutants,
field researcher to devise a sampling scheme
that allows a statistically robust estimate of species
cover. Mapping community cover, if considered
necessary to consider
common
wet deposition.
a
to assure repeatability
lichens
many lichens
tested in controlled
concentrations
often-diverse
of
however,
1983);
available
of
is some information
There
to determine their responses to different
studies.
Mapping Community Cover
Because of the varied habitats
using
Taxonomy Update
CONCERNS
expertise.
of
this element
Nriagu and Glooschenko (1992)
accumulation in mosses
in different locations in Canada.
collected and sent to a lab for analysis of
in the
the baseline chemical constituents
indicator species.
G) Transplants:
the source
has been raised by
during their study
E) Community Analysis: As part of the
floristic survey, Ryan described species
and their abundances.
Samples
and (3)
The possibility
air pollutants.
sulfur isotopes to identify
D) Indicator Species: Ryan identified
sensitive and tolerant species based on
other dose-response work. His analysis
showed the need for additional work on
the identification of indicator species.
F) Chemistry/Physiology:
substrate, and marine aerosols,
anthropogenic
in
combination
due to windblown dust,
124
of
from the fumigation
evidence
the
study, and the
may be persuasive.
—
Table 2.
Cost
Estimates
for Conducting
Various
Lichen
Studies.
COST ($)
(per unit)
# of SITES
METHOD
UNIT
(per veg type)
study
-
site
area
300
30+
all species
200-400
site
20-60+
all species
300-500
study
site
1-10 species
many species
20,000-50,000
10+
site
6-20
1-few species
1,000-3,000
sample
20-100
Site characterization
Floristics
Community
analysis
Indicator species
fumigation
gradient
Transplants
Chemical
analysis
SCALE
15,000-25,000
multiple elements/sample
analysis: 25-200
collection: 50-300
SUGGESTED FURTHER RESEARCH
COST ESTIMATES
The previous
chapters describe methods and
Table
quality.
2 gives a range
with each method.
method is applied,
the specific
factors
is historical,
costs associated
of
of
of
metals, nitrogen oxides,
level
of
detail
among methods or specifying
of
analysis
are not necessarily
very
possible
the
Rather, the scientifically best method(s) should be
chosen and scaled to an affordable level. Authors
in this manual
studies are worthless
state
our knowledge
pollution.
of
worth the cost.
1.
They also
about how many sites are needed, which
indicator
species are appropriate
found.
If
preliminary
a community
analysis
2.
and where they are
is planned, a
3.
study is essential to determine the
number and location
of
homogeneous
sites.
Costs
4.
can be cut by combining data collection efforts. For
example, floristics data, community
specimens can be collected
data and tissue
simultaneously
in
and
Howeve,r lichens can
of
pollutants;
it is
in lichen
with ambient
concentrations
There are many gaps in
about how lichens
react to air
We suggest the following areas for further
Fumigation studies to determine if there
and bioaccumulation
potential for specific pollutants: nitrogen
oxides, ozone, carbon dioxide, and toxic
air contaminants.
Video/photographic techniques that can be
used for community analyses.
are indicator species
emphasize that in most cases, a preliminary study
should be done before costs are estimated, to make
decisions
PAN,
research to begin to full those gaps:
that monitoring
unless they are done well, and
a good study is usually
to measure pollutant
air pollution concentrations.
program.
a monitoring
to
to which lichens
tissues and correlate this information
Cost should not be the sole or even the major
the chapters
sensitive.
still be used as accumulators
to be applied.
determinant for developing
radionculeides,
ozone are major modern pollutants
rough estimates and should be used only as a guide
when choosing
sensitive
and most air pollution was
by SO2 emissions earlier in this century
and in the last century. Organic air toxics, heavy
area,
projected costs.
The costs presented here should be considered
to air pollution
accompanied
the project
the study, and many other
result in a wide range
since lichens are mostly
SO2 pollution,
The detail with which each
the remoteness
objectives
will
of
The idea that lichens are sensitive
of air
techniques for using lichens as bioindicators
Standard
methods
and
QA/QC
procedures
for chemical analyses.
Development of herbaria/libraries of useful
lichen species that are accessible to
researchers and relevant to different
habitats.
the
field.
5.
125
Development of easy-to-use field and
laboratory guides to lichen taxonomy that
can be used for routine floristic surveys
and are kept up-to-date as new species
are identified.
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Increasing Point Source Emissions
Garry, J. 1987.
Constant Point Source Emissions
(De Not.) Bagl. transplanted at
biomonitoring sites around a new coal-fired
L.K Thompsoa 1980. Lichens as
of fluoride emissions from a phosphorus
Roberts, B.A;
indicators
plant, Long Harbour,
Newfoundland,
Journal of Botany
Canadian
L.K.
Roberts, B.A.;
emissions
from a phosphorus
Newfoundland,
Botany
as indicators
Canada.
Research 43:
Canada.
of
17:175-183.
Fuchs, C.; J. Garty. 1983.
of
Ramalina
lichen
57:1583-1590.
Sidhu, S.S.; B.A. Roberts.
of
vegetation in the vicinity
a phosphorus
Long Harbour, Newfoundland, Canada. Canadian
Forest Service, Bi-monthly Research Notes,
Environment
content
of
Fluorine
1978.
of
a
Ann. Bot. Fenn. 15: 158-166.
B.A.; S.S. Sidhu; B.A. Roberts. 1979.
in soil and vegetation in
Thompson,
Fluoride
D.I.; F.N. Haynes. 1973. Distribution
of some epiphytic lichen around an oil refinery at
Fawley, Hampshire, Chapter 5, In Ferry, B.W., M.S.
Baddeley and D.L. Hawksworth, (Eds), Air
Pollution and Lichens. London, Athlone Press of the
Morgan-Huws,
H. Olkkonen.
two lichen species in the vicinity
fertilizer factory.
accumulation
vicinity of a phosphorus plant
Pollution 18:221-234.
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of London.
University
the
Skye, E. 1968.
Environmental
A.; M.R.D. Seaward. 1979.
Monitoring lichen reinvasion of ameliorating
Henderson-Sellers,
Seaward, M.R.D.
R
Schubert
1980.
of
bioindicators
Pollution,
19:207-213.
and Schuh
J
environments.
Lawrey,
1975a.
p.
Uppsala,
Sweden,
123 pp.
1988.
Lichen evidence for
Park.
The Bryologist 9: 21-23.
Decreasing Regional Emissions
17-23.
Coppins, B.J. 1973. The "Drought
Ferry, B.W.; Baddeley,
The foliose and fruticose lichen
Hypothesis",
M.S.; Hawksworth,
(Eds), Air Pollution and Lichens,
of around the Muskingum River Power Plant,
Morgan Count, Ohio, Ohio Journal of Science,
London,
75:33-35.
Athlone
Press
of
In
D.L.
pp. 124-142,
the University
of London.
D.L.; P.M. McManus. 1989. Lichen
in London under conditions of rapidly
falling sulphur dioxide levels, and the concept of
zone skipping. Botanical Journal of the Linnean
Hawksworth,
R.E. 1975b. Lichens
quality around a coal-fired
Bryologist,
Showman,
in the
Acta Phytogeerographica
J.D.; M.E. Hale.
National
In:
flora
Showman,
- a study of
changes in atmospheric pollution in Shenandoah
Martin-Luther-Universitat Halle-Wittenberg.
R.E.
Air pollution
(eds.): Bioindikation auf
der Ebene der Individuen,
Showman,
and
Increasing Regional Emissions
The use of lichens as
ameliorating
region.
Suecica, 52.
Decreasing Point Source Emissions
Environmental
Lichens
epiphytes and environment
cryptogamic
Stockholm
environments.
(De Not.) Bagl. at air
stations. Environmental
New Point Source Emissions
Canada 31: 41-42.
Takala, K.; P. Kauranen;
content in the
Botany 23:29-43.
Experimental
plant.
Elemental
duriaei
quality biomonitoring
Damage to
1975.
(De Not.) Bagl. transplanted in
stations. Water, Air, Soil Pollution
biomonitoring
plant, Long Harbour,
Canadian Journal
Heavy metals in the lichen
1982.
duriaei
Ramalina
fluoride
power
Environmental
operation.
104-116.
Garty, J.; C. Fuchs.
S.S. Sidhu. 1979.
Thompson;
bryophytes
of
station after 1 year
58: 2218-2228.
Terrestrial
Metal amounts in the lichen Ramalina
duriaei
as indicators
of air
recolonization
power generating plant.
78:1-6.
R.E.
1981.
Lichen recolonization
air quality improvement.
Bryologist,
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following
D.L.; F. Rose. 1970. Qualitative scale
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Hawksworth,
84:492-497.
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227:145-148.
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JR.
Laundon,
Naturalist
Rydzak,
J.
London's lichens.
1970.
Ryan, B.D. 1990
London
49: 20-69.
1969.
Marble
Lichens
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conditions
ecological
Universitatis
Mariae
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of
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Curie-Sklodowska,
Baseline Study.
the
San Francisco,
Annales
CA,
46 pp.
Concerns
Nriagu, J.O.; W. A. Glooschenko.
Lichens and Air Quality In
Areas in California: A Series of Baseline
Ryan, B.D. 1990
composition
Environ.
Report to the U.S. Forest Service, San
CA, 28
Report to the U.S. Forest Service,
CA,
54 pp.
Clean Air Areas
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the
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Report to the U.S. Forest Service, San Francisco,
289:
289-292.
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In
Quality
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Lichens and Air Quality In
A Series of Baseline
Studies. Supplement, Element Analysis of Lichens.
Rose, C.I.; D.L. Hawksworth.
Wilderness
Air
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Wilderness
1981.
Lichen
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and
Wilderness
Ryan, B.D. 1990
C,
23:131-164.
recolonization
Lichens
Mountains
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Scien. and Technol.
Sigal, L.; T. Nash. 1983.
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1992.
Isotopic
sulfur in mosses across Canada.
26:
85-89.
Lichen communities
on
conifers in southern California mountains: an
ecological
Ecology
127
survey relative to oxidant air pollution.
64:
1343-1454.
Appendix I: List of
Participants
William Anderson
National Park Service
National Capital Region
1100 Ohio Drive, S.W.
Washington, D.C. 20242
Jesse Ford
Oregon
University
State
Department
of
Fisheries
Tamara Franklin-Blett
U.S. Forest Service
P.O. Box 25127
Lakewood, CO 80225
Jayne Belnap
National Park Service
125 W. 2nd South
Moab, Utah 84532
Linda Geiser
U.S. Forest Service
P.O. Box 309
James Bennett
National Park Service CPSU
University of Wisconsin
1040 WARF Building
610 Walnut St.
Madison, WI 53705
Petersburg,
AK
99833
Larry Gough
U.S. Geological Survey
Box 25046, MS 973
Charis Bratt
Denver Federal
Santa Barbara
and Wildlife
Corvallis, OR 97333
Museum of Natural History
Center
CO 80225
Denver,
2559 Puseta Del Sol Rd.
Santa Barbara,
CA
Larry Jackson
U.S. Geological Survey
Box 25046, MS 973
93005
William Denison
Dept.
of
Oregon
and Plant Pathology
Botany
Denver Federal
State University
Center
CO 80225
Denver,
Corvallis, OR 97331
Martha Makholm
Wisconsin Dept. of Natural Resources
P.O. Box 7921
Robert Doty
Air Quality
Program
Manager
U.S. Forest Service
Madison,
WI
53707
630 Sansome St.
San Francisco,
CA
94111
Deborah
Mangis
EPA-AREAL
Mail Drop 75
RTP, NC 27711
Sharon Eversman
Biology Department
Montana State University
Bozeman,
MT
59717
Tonnie
Air
Rich Fisher
U.S. Forest Service
240 W. Prospect St.
Ft. Collins, CO 80526
Maniero
Division
Quality
National
Park Service
P.O. Box 25287
Denver, CO 80225
128
Bruce McCune
of
Department
Bruce Ryan
General
Science
Fremont
National
Forest
P.O. Box 25
Bly, OR 97622
State University
Oregon
Weniger Hall 355
Corvallis, OR 97331
David Schwartzman
Will Moir
Department
U.S. Forest Service
240 W. Prospect St.
Ft. Collins, CO 80526
Howard University
Washington, D.C. 20059
of
and Geography
Geology
Ray Showman
American Electric Power Service Corp.
Barbara Murray
University of Alaska Museum
Fairbanks,
AK
99775
of
Botany
Tom Nash
Lorene
Department
AZ
85281
Lorenz Pearson
Biology Department
Rick's College
Rexburg,
ID
Sigal
Clifford Smith
Park Service
National
CPSU
of Botany
University of Hawaii
Department
83460
3190 Maile Way
Honolulu, HI 96822
Sherry Pittam
Botany
OH 43215
Oak Ridge National Laboratory
P.O. Box 2008
Oak Ridge, TN 37831
and Microbiology
Arizona State University
Tempe,
Plaza
1 Riverside
Columbus,
/NHB
166
Smithsonian
Institute
Larry
Washington,
D.C. 20560
1076
Clair
St.
West 550 South
Orem,
UT
84058
Fred Rhoades
4320 Dumas Avenue
Kenneth
WA
Bellingham,
Stolte
U.S. Forest Service
Southeast Forest Experiment
Al
P.O. Box 12254
RTP, NC 27709
Riebau
Colorado
State
2401 Research
University/BLM
Blvd.
Mike Treshow
Biology Department
Suite 205
Fort Collins, CO 80526
University
of
Utah
Salt Lake City,
Barry Rock
UT
84112
Science and Engineering Research
University of New Hampshire
Durham, NH 03824
Clifford Wetmore
Botany
Roger Rosentrcter
Bureau of Land Management
Idaho State Office
3380 Americana
Boise,
ID
Department
University
St. Paul,
Maggie
of Minnesota
MN 55108
Wilson
Tetra Tech, Inc.
Terrace
10306 Eaton PI. Suite 340
83706
Fairfax,
129
VA
22030
Station
Appendix II: Reviewers
Steve Link
Jayne Belnap
National
Canyonlands
Battelle Northwest
Parte
125 W. 2nd South
Moab, UT 84532
P.O. Box 999
Bob Egan
Dept. of Biology
University of Nebraska
Tonnie
Omaha,
Randy
NE
99352
Maniero
Air Quality Division
National
at Omaha
Park Service
P.O. Box 25287
Denver, CO 80225
68182-0040
Ferrin
Air Quality Division
Tom Moeser
c/o U.S. EPA Environmental
200 SW 35th St.
Corvallis, OR 97333
National Park Service
P.O. Box 25287
Denver, CO 80225
Research
William Moir
Rich Fisher
USDA
WA
Richland,
Forest
USDA
Service
Station
Experiment
Forest Service
Rocky Mountain Forest and
Rocky Mountain Forest and Range
Station
Range Experiment
240 W. Prospect
240 W. Prospect
Fort Collins, CO 80526
Fort Collins, CO 80521
JoAnn Flock
Bob Musselman
University of Colorado Herbarium
Campus Box 350
Boulder, CO 80309
USDA
Forest Service
Rocky Mountain Forest and Range
Experiment
Station
240 W. Prospect
Fort Collins, CO 80526
Linda Geiser
USDA
Forest
Stikine
Area
#15
Service
Louise O'Deen
USDA
12th St.
Petersburg,
AK
Forest Service
Rocky Mountain Forest and Range
P.O. Box 309
99833
Experiment
Station
240 W. Prospect
Fort Collins, CO 80526
Mark Jensen
USDA
Forest
Service
Phil Rundel
UCLA-Laboratory
Northern Region
Building
P.O. Box 7669
Missoula, MT 89807
Region
1,
Federal
Environmental
of Biomedical
Science
900 Veteran Ave.
Los Angeles,
130
CA
90024
and
Lab
Susan Sater
Sam Shushan
USD A Forest Service
Pacific NW Region
533 SW First Ave.
P.O. Box 3623
2010 Mariposa Ave.
Portland,
OR
Boulder,
CO 80309
John Thompson
Dept. of Botany
97208
University
Mark Scruggs
Air Quality Division
National Park Service
P.O. Box 25287
Denver, CO 80225
Madison,
131
of Wisconsin
WI 53706
Descriptions of back-cover photos, clockwise from the upper left corner:
Bl.
Lobaria oregana, found on tree branches and
winter forage for deer and mountain
trunks, provides
goats. Internal
cephalodia
contribute
amounts of fixed nitrogen.
Collected for tissue
in Tongass Air Quality Biomonitoring
Stephen
B2.
twin, P. elegantula
(Zahlbr. ) Szat.
to
Melanella
by
(transferred
Essl.) are not sufficiently
to be useful as indicators; they are
discriminating
exasperatula
and they found it exasperating.
B3.
Hypogymnia
B5.
Photo by
Hypogymnia
enteromorpha
(Gough et. al. 1988).
enteromorpha
(Ach.) Nyl. in
and Usnea spp. in the background
by-
(Ach.) Nyl. This
species was used to measure baseline element
concentration ranges in Redwood National Park
1990,
Lorenz Pearson.
foreground
Photo
Cladina rangiferina, reindeer or caribou
This and other Cladina species are the
primary food of caribou and occasionally of people
C. rangiferina is common
during times of famine.
This species was
among mosses in muskegs.
collected for tissue analysis by Tongass Air Quality
Biomonitoring Project. Photo by Sylvia Duran
Sharnoff and Stephen Sharnoff.
P.
was used by Rope and Pearson
Park.
lichen.
Parmelia exasperatula Nyl. on Juniperus
bark. This species and its nearly
also difficult to remove from the bark.
National
B4.
Photo by Sylvia Duran Sharnoff and
Sharnoff.
scopulorum
identical
ranges in Redwood
Larry Jackson.
significant
analysis
Project.
used to measure baseline element concentration
B6.
the
on
Usnea spp.
Photo by Larry Jackson.
This species was used
measure baseline element concentration
Douglas-fir in the Little Bald Hills ultramafic region
of Redwood National Park. These two species were
USD A policy prohibits discrimination
Redwood
by
National
Park (Gough
to
ranges in
et. al 1988).
Photo
Larry Jackson.
because of race, color, national
condition. Any person who
believes he or she has been discriminated against in any
USDA-related activity should immediately contact the Secretary of
Agriculture,
Washington, DC 20250.
origin, sex, age religion,
or handicapping
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