URGE Application
- Cover Sheet
Deadline: January 27, 2011
Must be typed
College or University: Western Michigan University
Student 1
Name:
Student #:
Local Address:
Permanent Address:
Phone:
E-mail:
Major: Biology
Emphasis (if applicable): Environmental Science
Anticipated graduation date: Summer 2011
Current GPA: 3.49
Student 2 (if applicable)
Name:
Student #:
Local Address:
Permanent Address:
Phone:
E-mail:
Major: Environmental Studies and Geography Emphasis (if applicable): Environmental Resource Management
Anticipated graduation date: May 2012
Current GPA: 3.94
Faculty
Name: Stephen B. Malcolm
Department: Biological Sciences
Address: 3151 Wood Hall, Western Michigan University, 1903 West Michigan Avenue, Kalamazoo, MI 49008
Phone: 269-387-5604
E-mail: steve.malcolm@wmich.edu
Preferred Meeting topic: GIS/niche modelling
1. Title of Research Project: Temporal and spatial changes in monarch butterfly population synchrony in Michigan
and density-dependent influences on the incidence of the protozoan parasite, Ophryocystis elektroscirrha
Dates of Project: 20 May 2011 to 30 September 2011
Institute On-site Housing and Meal Support (Estimated number of days on property)
Faculty: 20 days 5 overnights
Student(s): 60 days 60 overnights
2. Does this proposal require approval from:
Human Research Review Committee
Institutional Animal Care and Use Committee
(If either of these is required, a copy of the approval must be submitted to Pierce Cedar Creek Institute.)
3. Budget Summary
a. Student Stipend Request
$7,000 (not to exceed $3,500 per applicant)
b. Faculty Stipend Request
$1000
Project Support (i.e. supplies, equipment, travel, etc):
Total
$2,500 (Faculty stipend and project support not to
exceed $3,500 combined)
$10,500 (not to exceed $10,500)
Signatures
_________
Student Researcher
________
Faculty Mentor
1/26/2011
Date
1/26/2011
Date
________
Student Researcher
_
Department Head
1/26/2011
Date
________
Date
URGE Application
1. Proposal: Temporal and spatial changes in monarch butterfly population synchrony in Michigan and
density-dependent influences on the incidence of the protozoan parasite, Ophryocystis elektroscirrha
a. Synopsis:
We propose to determine how synchrony of monarch population density in southwest Michigan changes with
time and the influence that such changes might have on the incidence and prevalence of a protozoan pathogen specific to
monarch butterflies. While much is known about monarch migration, reproductive behavior, hostplant use and defense
against natural enemies, we know little about the most important phase of monarch life history when most population
increase is thought to occur on the common milkweed, Asclepias syriaca, in the Great Lakes region.
b. Introduction:
The monarch butterfly, Danaus plexippus, is an iconic example of insect migration by virtue of its predictable,
long-distance annual migration in North America between overwintering locations in Mexico and breeding habitat
distributed across the USA and southern Canada east of the Rocky Mountains (Figure 1; Urquhart, 1960; Johnson, 1969;
Urquhart and Urquhart, 1976, 1978; Baker, 1978; Brower, 1985; Calvert and Brower, 1986; Malcolm, 1987; Brower and
Malcolm, 1991; Brower, 1995, 1996).
Figure 1. Distributions of monarch butterfly overwintering locations in Mexico (A) and 4 common
milkweed larval hostplants (Asclepias spp.) in North America east of the Rocky Mountains (Malcolm et
al., 1993), with a photograph of 2 adult monarchs basking on leaves at the Sierra Chincua overwintering
location near Angangueo, Michoacán, Mexico (photograph by L.P. Brower).
From early wing-tagging studies by Nora and Fred Urquhart in Canada (Urquhart, 1960; Urquhart and Urquhart,
1976, 1978) and later studies coordinated by Orley Taylor of Monarch Watch at the University of Kansas
(http://www.monarchwatch.org/index.html) we know that adult monarchs fly each autumn to 12 locations in oyamel fir
forests above 3,000 m altitude in the Sierra Transvolcanica mountains of central Mexico, west of Mexico City (Calvert
and Brower, 1986; Brower, 1985, 1995, 1996). There the butterflies remain for up to 5 months over winter in tightly
aggregated clusters until their return migration north in spring at the end of March. We also know that the same
butterflies that left the USA and southern Canada in the autumn, and spent the winter in Mexico, return to the southern
USA where they mate, lay eggs and die (Cockrell et al., 1993; Malcolm et al., 1993). The offspring of these migrants
from Mexico then feed on southern hostplant species and upon emergence as adults continue the migration north to
recolonize the full spatial extent of their milkweed hostplants across North America, east of the Rocky Mountains
(http://www.learner.org/jnorth/monarch/index.html).
In addition to their spectacular annual migration in North America, monarch butterflies are well known for their
specialized larval feeding on milkweeds in the genus Asclepias and their ability to sequester toxic steroids known as
cardenolides for use in defense against natural enemies such as birds (Brower et al., 1967, 1968, 1972; Brower, 1969;
Brower and Moffitt, 1974; Fink and Brower, 1981; Malcolm and Brower, 1989; Glendinning and Brower, 1990;
Malcolm, 1990; Alonso-Mejia and Brower, 1994; Malcolm, 1995). Interestingly, each Asclepias hostplant species
generates a different pattern of sequestered cardenolides in adult monarchs and Malcolm et al. (1993) used these
chemical “fingerprints” as indicators of migratory strategy in spring migrants. Malcolm et al., hypothesized that
monarchs either migrate using a “single sweep” strategy or a “successive brood” strategy (Figure 2). In “single sweep”
migration the entire breeding distribution of monarchs would be accessed by overwintered migrants that move north and
lay eggs as they progress through both space and time. In “successive brood” migration the migrating spring butterflies
would reach the southern USA from Mexico, mate, lay their eggs on southern Asclepias species (Figure 1) and die,
leaving their offspring to continue the migration north. Evidence from both cardenolide fingerprints (Malcolm et al.,
1993) and isotopic signatures (Wassenaar and Hobson, 1998; Hobson et al., 1999) show that spring migration does
occur via successive broods.
While much is known about migration, mating behavior and physiology, hostplant use and the operation of
defense against natural enemies in monarch butterflies, we still know little about the dominant phase of their life history
when their annual populations increase during the summer. Michigan is in the center of the extensive distribution of the
common milkweed, Asclepias syriaca, which is the most important hostplant for monarchs (Malcolm et al., 1989), out
of the 108 species of Asclepias milkweeds known from North America (Woodson, 1954) and it is in their northern range
that monarchs generate up to three successive generations (Borkin, 1982; Malcolm et al., 1987), to establish the large
numbers of butterflies that fly to Mexico each autumn.
Figure 2. Hypothesized spring migration strategiesfor monarch butterflies that recolonize North American
milkweed breeding habitat east of the Rocky Mountains (Malcolm et al., 1993)
In addition to characteristic patterns of hostplant use facilitated by migration, monarch butterflies are also
subject to attack by a highly specific protozoan pathogen called Ophryocystis elektroscirrha and both incidence and
prevalence of the parasite vary through the annual cycle of monarchs (Altizer et al. 2000) and also with host plant
species (De Roode et al., 2008). However, nothing is known about possible changes in the incidence or prevalence of
this protozoan in relation to increasing density of monarchs across the three generations that characterize their life
history in the Great Lakes region (Malcolm et al., 1987).
Here we propose to research monarch breeding ecology during the summer throughout southwest Michigan in
order to determine (1) the changing nature of spatial and temporal synchrony of monarch generations through the
summer, and (2) the relationship between generation synchrony and both monarch density and the incidence of a
parasitic disease. Thus we propose the following null hypotheses and their associated alternate hypotheses:
Null hypothesis 1: Adult monarch arrivals to Michigan and their occurrence throughout the summer are random.
Alternate hypothesis 1a: Adult monarch arrivals to Michigan are highly synchronized and predictable
in May and June and the consequent incidence of larvae is initially highly synchronized.
Alternate hypothesis 1b: Both adult and larval monarch distributions lose synchrony in space and time
as the summer progresses.
Alternate hypothesis 1c: Synchrony is restored by changing temperature and daylength as the autumn
equinox is approached and monarchs leave Michigan in September.
Null hypothesis 2: The incidence of spores of Ophryocystis elektroscirrha on monarch adults and eggs in the
field is random and density independent and does not change with time.
Alternate hypothesis 2a: The incidence of O. elektroscirrha spores on monarch adult abdomens is
positively density dependent and increases with time through the summer.
Alternate hypothesis 2b: The incidence of O. elektroscirrha spores on monarch adult abdomens is
influenced by larval host plant species.
c. Methods:
1. Searching by Andrew Johnson, Joshua Armagost and Stephen Malcolm on a continuous cycle throughout
southwest Michigan for monarch adults, eggs and larvae from late May through to early September in a
variety of habitats, with an emphasis on Pierce Cedar Creek and surrounding farmland.
2. The milkweeds that occur in Michigan include the following Asclepias species ranked according to abundance:
1)
2)
3)
4)
5)
6)
7)
8)
9)
Asclepias syriaca (extremely abundant and highly modular)
A. incarnata (common)
A. tuberosa (common)
A. amplexicaulis (uncommon)
A. verticillata (uncommon but can be locally abundant)
A. exaltata (uncommon)
A. viridiflora (rare)
A. purpurascens (rare)
A. sullivantii (rare)
3. ARCview monarch mapping by Andrew Johnson of latitude and longitude coordinates for different time
intervals, collected from stored adult monarchs in US collections.
4. Field surveys of milkweed distribution and abundance in southwest Michigan and ARCview mapping to show
densities of different milkweed species and relative biomasses. Personal vehicles will be used by Joshua
Armagost, Andrew Johnson and Stephen Malcolm to search milkweeds over an area that stretches from Lake
Michigan east to Pierce Cedar Creek and south to Battle Creek, Schoolcraft (immediately south of Kalamazoo)
and Benton Harbor, including protected areas such as the Allegan State Game Preserve and Fort Custer.
5. ARCview mapping by Andrew Johnson of larval densities on milkweeds and adult observations from the field
and determination of statistical relationships for each milkweed species between milkweed and monarch
distributions and abundances (spatial and temporal synchrony determinations) using methods of Borkin
(1982), Malcolm et al. (1987) and Cockrell et al. (1993).
6. Joshua Armagost will perform abdomen and egg washes for Ophryocystis elektroscirrha spores and
comparison of data with the density data above using methods of Altizer et al. (2000) and De Roode et al.
(2008). The monarch abdomens and eggs will be from material located in the field associated with naturally
occurring milkweeds. Digital imaging and counting equipment is available in the Biological Imaging Center at
Western Michigan University.
7. Josh Armagost will perform rearing experiments with monarchs on common Michigan milkweeds and the
southern milkweed A. viridis at PCCI to determine possible effects on O. elektroscirrha. A. viridis will be used
because it is the primary host plant species used by migrant monarchs that arrive in Michigan in June
(Malcolm et al., 1993).
d. Research conduct:
Milkweeds are potentially toxic plants because they produce copious latex and their cardenolides target the Na +,
K+-ATPase enzymes of potential consumers, including humans (Malcolm, 1995). Thus care will be taken when
handling plants and insects by wearing disposable gloves. The protozoan pathogen is host restricted to monarch
butterflies and poses no risk to the researchers.
e. References:
Alonso-Mejia, A., and Brower, L. P. 1994. From model to mimic: age-dependent unpalatability in monarch butterflies.
Experientia 50:176-181.
Altizer, S. M., Oberhauser, K. S., and Brower, L. P. 2000. Associations between host migration and the prevalence of a
protozoan parasite in natural populations of adult monarch butterflies. Ecological Entomology 25:125-139.
Baker, R. R. 1978. The Evolutionary Ecology of Animal Migration. New York: Holmes and Meier.
Borkin, S. S. 1982. Notes on shifting distribution patterns and survival of immature Danaus plexippus (Lepidoptera:
Danaidae) on the food plant Asclepias syriaca. Great Lakes Entomologist 15:199-206.
Brower, L. P. 1969. Ecological chemistry. Scientific American 220:22-29.
Brower, L. P. 1985. New perspectives on the migration biology of the monarch butterfly, Danaus plexippus L.
Contributions in Marine Science Supplement 27:748-785.
Brower, L. P. 1995. Understanding and misunderstanding the migration of the monarch butterfly (Nymphalidae) in
North America: 1857-1995. Journal of the Lepidopterists' Society 49:304-385.
Brower, L. P. 1996. Monarch butterfly orientation: missing pieces of a magnificent puzzle. Journal of Experimental
Biology 199:93-103.
Brower, L. P., and Brower, J. V. Z. 1964. Birds, butterflies and plant poisons: a study in ecological chemistry. Zoologica
49:137-159.
Brower, L. P., and Malcolm, S. B. 1991. Animal migrations: Endangered phenomena. American Zoologist 31:265-276.
Brower, L. P., and Moffitt, C. M. 1974. Palatability dynamics of cardenolides in the monarch butterfly. Nature 249:280283.
Brower, L. P., Brower, J. V. Z., and Corvino, J. M. 1967. Plant poisons in a terrestrial food chain. Proceedings of the
National Academy of Sciences of the United States of America 57:893-898.
Brower, L. P., Ryerson, W. N., Coppinger, L. L., and Glazier, S. C. 1968. Ecological chemistry and the palatability
spectrum. Science 161:1349-1350.
Brower, L. P., McEvoy, P. B., Williamson, K. L., and Flannery, M. A. 1972. Variation in cardiac glycoside content of
monarch butterflies from natural populations in eastern North America. Science 177:426-429.
Calvert, W. H., and Brower, L. P. 1986. The location of monarch butterfly (Danaus plexippus L.) overwintering colonies
in Mexico in relation to topography and climate. Journal of the Lepidopterists' Society 40:164-187.
Cockrell, B.J., S.B. Malcolm and L.P. Brower. 1993. Time, temperature, and latitudinal constraints on the annual
recolonization of eastern North America by the monarch butterfly. Pages 233-251 In, S.B. Malcolm and M.P.
Zalucki (editors), Biology and Conservation of the Monarch Butterfly. Natural History Museum of Los Angeles
County, Science Series 38, 425 pp.
De Roode, J. C., Pedersen, A. B., Hunter, M. D., and Altizer, S. 2008. Host plant species affects virulence in monarch
butterfly parasites. Journal of Animal Ecology 77:120-126.
Fink, L. S., and Brower, L. P. 1981. Birds can overcome the cardenolide defence of monarch butterflies in Mexico.
Nature 291:67-70.
Glendinning, J. I., and Brower, L. P. 1990. Feeding and breeding responses of five mice species to overwintering
aggregations of the monarch butterfly. Journal of Animal Ecology 59:1091-1112.
Hobson, K. A., Wassenaar, L. I., and Taylor, O. R. 1999. Stable isotopes (δD and δ13C) are geographic indicators of
natal origins of monarch butterflies in eastern North America. Oecologia 120:397-404.
Johnson, C. G. 1969. Migration and dispersal of insects by flight. London:. xxii + 763 pp.
Malcolm, S. B. 1995. Milkweeds, monarch butterflies and the ecological significance of cardenolides. Chemoecology
5/6:101-117.
Malcolm, S. B., and Brower, L. P. 1986. Selective oviposition by monarch butterflies (Danaus plexippus L.) in a mixed
stand of Asclepias curassavica L. and A. incarnata L. in south Florida. Journal of the Lepidopterists' Society
40:255-263.
Malcolm, S. B., and Brower, L. P. 1989. Evolutionary and ecological implications of cardenolide sequestration in the
monarch butterfly. Experientia 45:284-295.
Malcolm, S. B., Cockrell, B. J., and Brower, L. P. 1987. Monarch butterfly voltinism: Effects of temperature constraints
at different latitudes. Oikos 49:77-82.
Malcolm, S. B., Cockrell, B. J., and Brower, L. P. 1989. The cardenolide fingerprint of monarch butterflies reared on the
common milkweed, Asclepias syriaca L. Journal of Chemical Ecology 15:819-853.
Malcolm, S.B., B.J. Cockrell and L.P. Brower. 1993. Spring recolonization of eastern North America by the monarch
butterfly: successive brood or single sweep migration? Pages 253-267 In, S.B. Malcolm and M.P. Zalucki
(editors), Biology and Conservation of the Monarch Butterfly. Natural History Museum of Los Angeles County,
Science Series 38, 425 pp.
Urquhart, F. A. 1960. The monarch butterfly: International traveler. Toronto: University of Toronto Press. 232 pp.
Urquhart, F. A., and Urquhart, N. R. 1976. The overwintering site of the eastern population of the monarch butterfly
(Danaus p. plexippus; Danaidae) in southern Mexico. Journal of the Lepidopterists' Society 30:153-158.
Urquhart, F. A., and Urquhart, N. R. 1978. Migrations of the eastern population of the monarch butterfly in North
America to the overwintering site in the neo-volcanic plateau of Mexico. Atalanta (Munnerstadt) 9:133-139.
Wassenaar, L. I., and Hobson, K. A. 1998. Natal origins of migratory monarch butterflies at wintering colonies in
Mexico: new isotopic evidence. Proceedings of the National Academy of Sciences of the United States of
America 95:15436-15439.
Woodson, R. E. Jr. 1954. The North American species of Asclepias L. Annals of the Missouri Botanical Garden 41(1):1211.
f. Timetable:
20 May to 30 September 2011.
dates
March - May 2011
20 May - 20 June
15 - 20 June
20 June - 20 August
21-31 August
October
April 2012
activity
start growing Asclepias viridis from seed in WMU glasshouses
searches across SW Michigan for first arrival monarchs
place potted milkweeds in common garden plot at PCCI
students resident at PCCI
Prepare presentation for wrap-up meeting in September
submit final research reports.
Present results at Midwest Ecology and Evolution Conference
g. Budget:
Item
1. Hausser bright-line hemacytometer
2. Hemacytometer cover glass
3. Insect nets (3)
4. Collapsible field cages (10)
5. Plant growth supplies (pots + medium)
6. Waterproof field notebooks (3)
7. All weather pens (3)
8. Mileage (3 cars x 1,000 miles x $0.51)
Source
Fisher Scientific
Fisher Scientific
BioQuip
BioQuip
WMU glasshouses
BioQuip
BioQuip
mileage
total
cost
$210.00
$22.00
$114.00
$500.00
$70.00
$24.00
$30.00
$1,530.00
$2,500.00
Budget justification:
1 and 2. A single hymacytometer and cover slips are required to count spores of Ophryocystis elektroscirrha
that are washed from the abdomes of wild-caight monarch butterflies (both males and females) and the
eggs found on milkweeds in the field.
3. Insect nets (18” diameter) are required to collect adult monarchs across SW Michigan and sample for
O. elektroscirrha spores.
4. Field cages are required to enclose feeding monarch larvae on native Asclepias species and on the
southern milkweed, Asclepias viridis, at PCCI to examine O. elektroscirrha spore impact on growing
larvae and the influence of host plant.
5. Pots and soilless medium are required to grow milkweeds.
6. Notebooks required to record field data for larval and adult presence from May to September.
7. Waterproof pens needed for data recording.
8. Much of the project will be spent driving on multiple occasions to the same milkweed locations to
sample for monarchs through time from approximately Saugatuck on Lake Michigan east to PCCI,
south to Battle Creek and west to Benton Harbor.
a. Plan for the presentation or dissemination of results:
We propose to prepare results for presentation at the annual Midwest Ecology and Evolution
Conference planned for April 2012, such as the meeting in Carbondale, Illinois to be held April 1-3, 2011
(https://www.dce.siu.edu/index.php/Conferences/2011-Midwest-Ecology-and-Evolution-Conference). We
also wish to post the results of undergraduate research on our departmental website at Western Michigan
University (http://www.wmich.edu/bios/) as part of our university mission of a “student centered research
university.”
b. Statements of commitment:
Both Josh Armagost and Andrew Johnson will be responsible for field searches and collection of
monarch adults, eggs and larvae. Josh Armagost will take primary responsibility for the Ophryocystis
elektroscirrha research and Andrew Johnson will take primary responsibility for all GIS mapping using
ARCview for which he has the necessary skills. Steve Malcolm will act as mentor throughout all phases of
the research and will be resident as necessary at PCCI to help with experimental work. In May and June Steve
Malcolm will spend 2 weeks in the field with the students to train them in milkweed searching techniques and
the identities of different milkweeds. Steve Malcolm has 28 years of experience working with monarch
butterfly migration and host plant use and the chemical ecology of defense. Both Josh Armagost and Andrew
Johnson are fully committed to this research and have no other commitments. Steve Malcolm has applied for
funds for a 2-week research trip to Argentina to work on the southern monarch butterfly, Danaus erippus, in
July, but otherwise he will be available in Kalamazoo and PCCI and for field searches in southwest Michigan
throughout the project.
As indicated above we plan to present results of this research at the annual Midwest Ecology and
Evolution Conference planned for April 2012, but as yet we do not know the venue for this annual event at
which student ecological and evolutionary research is showcased.
Steve Malcolm has no other commitments from May 20 until 3 September and is not teaching during
the summer.
9. Attachments
a. Student’s unofficial transcript reflecting courses completed and grades received.
b. Student’s one-page Resume (work, educational, volunteer, or other experiences that support the
student’s ability to carry out this project).
c. Letter of recommendation from the faculty mentor for the student researcher(s).