NA09NOS4260242
: 9/1/09 to 8/31/10
Phase shifts on coral reefs have been reported with increasing frequency worldwide over the past 40 years. These benthic communities shifted to unusually low levels of both coral and turf algae cover coincident with a persistent dominance of macroalgae. On
Hawaiian reefs this problem has been exacerbated further by introduced macroalgae.
These invasive species have replaced not only coral and turf algae but also native seaweeds, such that the benthic cover of some Hawaiian reefs is dominated by species unknown in the islands <60 years ago. The green alga Avrainvillea amadelpha
(Bryopsidales) is one example of this phenomenon. First reported in 1981, A. amadelpha has become the dominant cover across a wide range of water depths and substrate types. The coral reefs of Kalaeloa, on the Leeward Coast of Oahu, are located close to the first report of this species on Oahu. For about 15 years scientists at the US Fish and
Wildlife Service have been observing this area, concerned about the increasing presence of A. amadelpha on the Kalaeloa reefs. This study was undertaken to provide managers with baseline information on A. amadelpha within subtidal, hard bottom communities of Kalaeloa. We took a two-fold approach to understanding the distribution of A. amadelpha . First, we conducted rapid ecological assessments of sites across the area of interest. The results provided a spatial contrast of habitat characteristics for invaded versus non-invaded habitats. Then, we conducted experimental manipulations to test whether grazer assemblages contribute to the distribution of the standing crop of
A. amadelpha recorded at Kalaeloa.
A.
Detailed description of the resource management problem(s) to be addressed . This project was requested to address the status of an invasive alga Avrainvillea amadelpha around the coral reefs of Kalaeloa, Oahu. Resource managers have reported that this species has been increasing in coverage around this site over the last 15 years and they requested that we map this species’ distribution and characterize the invasive species in this area.
B. Overarching goal(s) of the project : Characterize the marine habitats around Kalaeloa
– Campbell Industrial Park colonized by the invasive alga Avrainvillea amadelpha .
C. Detailed description of the question(s) asked to answer the resource management problem(s)
A.
Where is A. amadelpha found around Kalaeloa? Provide GPS data on its distribution as baseline information on this species. Are there any patterns associated with A. amadelpha distribution at Kalaeloa?
B.
Is the physical removal of A. amadelpha a management option for this site?
Is there any alternative management options to be considered?

C.
What habitats have been invaded by A. amadelpha ?
D.
Because excessive nutrient loads in coastal waters have been linked to algal invasions elsewhere, is there any evidence that nutrient loading is contributing to the invasion of A. amadelpha around Kalaeloa?
E.
Is there any evidence of heavy metals accumulation by A. amadelpha around
Kalaeloa?
Objectives to answer each question
A. Map the areal extent of the invasive alga Avrainvillea amadelpha along the coastline from Kalaeloa
B. Monitor the fate of open gaps created by the physical removal of invasive algal biomass
C. Quantify and compare water quality inside and outside Avrainvillea amadelpha canopies
D. Quantify stable isotopes accumulated by Avrainvillea amadelpha canopies
E. Quantify and compare C:N:P of Avrainvillea amadelpha canopies
F. Quantify and compare heavy metals accumulated by Avrainvillea amadelpha canopies
G. Organize a workshop for managers to discuss the results and next steps
A. Map the areal extent of the invasive alga Avrainvillea amadelpha along the coastline from Kalaeloa. Are there any patterns associated with Avrainvillea
amadelpha distribution at Kalaeloa? i. List individuals and organizations actually performing the work: F. Thomas and K. Peyton planned the work; K. Peyton and K. Groom collected data in the subtidal sites; K. Peyton and L. Valentino collected data in the intertidal sites; L. Valentino assisted on the dive boat; K. Peyton analyzed the data. ii. Material list: photoquad; digital cameras with underwater housings; underwater strobe; chain for estimating habitat heterogeneity index; transect tape; GPS; ruler; collection bags; underwater paper; clipboard; 10 cm 2 quadrat iii. Construction instructions for anything used to accomplish the III(D) objectives: photoquads see Preskitt et al. (2004); habitat heterogeneity index see Risk (1972) iv.
o For sites sampled by scuba diving, two independent 15 m transects were laid perpendicular to the shore (shore normal) o Water depth o Latitude/longitude (GPS unit towed on a surface float) o Habitat type:
 Deep reef (along15-20 m escarpment)
 Pavement near reef <30 m from reef structure
 Pavement >30 m from reef structure
 Shallow reef (inshore 6-12 m)
 Seagrass meadow o Benthic contour complexity/habitat heterogeneity index = measure of fine-scale habitat heterogeneity (Risk 1972) o Urchin counts (along 15 x 2 m belt transect measuring urchin density by species)

o Fish species checklist (record species observed) o Percent cover of functional groups (coral; turf algae: coralline algae; invasive algae; urchins; etc.) was estimated from 6 photoquad images taken at random points along each transect (Preskitt et al. 2004) o Within the belt transect area the target species, A. amadelpha, was recorded as either present or not observed. Additionally, on the dives this species was observed at the site but not within a belt transect area, it was noted as rare. When A. amadelpha was present within a belt transect area its standing crop was estimated as:
 Biomass per unit area (blotted wet weight for some samples and dry weight for all samples) collected by hand from a 10 cm 2 quadrat (n = 3-5 per transect)
 In the laboratory biomass per unit area was partitioned as:

Total host biomass ( A. amadelpha )

Epiphyte biomass (removed by scraping with a glass slide)

Total host + epiphyte biomass

Host frond biomass

Host holdfast biomass

Percentage of fronds with grazing scars
 Dry weight was recorded once the samples reached a constant weight after being placed in a drying oven at 60 o C
(48-72 hours)
 A. amadelpha canopy height was estimated using a ruler and measuring to the nearest 0.5 cm v. Data analysis techniques: Coral Point Count; ANOVA vi. Photos of the research at each stage:
Photoquad with diver changing numbered tags that correspond to transect and photo number.
Example – screen capture of photoquad image in
Coral Point Count.
B. Monitor the fate of open gaps created by the physical removal of invasive algal

biomass. Is the physical removal of the invasive species an option? Is there an alternative approach ? i. List individuals and organizations actually performing the work: F. Thomas and K.
Peyton designed the experiment; K. Peyton and L. Valentino constructed the cages and tethers; K. Peyton, L. Valentino and K. Groom set up the experiments with K.
Peyton and K. Groom doing the deployment. K. Peyton analyzed the data. ii. Construction instructions for anything used to accomplish the III(D) objectives

Cages were constructed out of plastic coated metal 1.3 cm 2 mesh and measured 15 x 9 x 10 cm

Cable ties were used to seal the cages

Tethers were constructed out of twisted polypropylene rope

Cages and tethers were anchored to the substrate using cable ties iii. Deployment steps

Collect A. amadelpha ramets from an area with a mature canopy and place in mesh bags

Rinse ramets in saltwater to remove sediments and dislodge mobile invertebrates

On land (stable platform), recorded a blotted wet weight for each replicate

Replicates were randomly assigned to treatments and sites

Set out all replicates on the same day in two habitats: collection site (=donor site) and coral reef site (= grazers)

For each habitat A. amadelpha was tethered in caged and uncaged treatments to obtain an estimate of grazing intensity (=difference between the specific growth rates of caged and uncaged)

Sample size was six

One preliminary experiment was run to estimate the approximate grazing rate and determine each experiment ’s duration

For the first experiment the duration was 2 days

Second experiment was conducted as described above with the exception that only the donor site was used

For the second experiment the duration was 68 days iv. Data collection procedures

Alga was blotted and weighed

Percent change in biomass was calculated for each replicate v. Data analysis techniques

Student t-test and ANOVA vi. Photos from research during each stage (construction, in situ, lab)

C. Quantify and compare water quality inside and outside Avrainvillea amadelpha canopies i. List individuals and organizations actually performing the work: K. Peyton and K.
Groom did the diving. K. Peyton collected samples.
D. Quantify stable isotopes accumulated by Avrainvillea amadelpha canopies i. List individuals and organizations actually performing the work: F. Thomas and K.
Peyton designed the experiment; K. Peyton collected the samples; K. Peyton and
L. Valentino processed samples; Popp Lab (SOEST) analyzed samples ii. Material list:

Balance

Drying oven (60 o C)

Wig-L-Bug amalgamator using stainless steel liners inside threaded screw vials (plastic)

Scintillation vials

Desiccator iii. Data collection procedures

Samples A. amadelpha were collected from each site and transported in seawater to laboratory

Because A. amadelpha was not observed at each site, a second macroalga found at all sites, Neomaris vanbosseae, was added to the study. Where both species co-occurred they were collected together.

Samples were collected in triplicate

At the lab, all samples of N. vanbosseae were scraped with a glass slide to remove epiphytes

A. amadelpha was divided in three sample types o A. amadelpha - new fronds with no visible epiphytes o Whole plant A. amadelpha – all fronds with epiphytes o Whole plant A. amadelpha – holdfast

Rinsed samples in Nanopure water to remove sediments and salts

Samples were dried to a constant weight at 60 o C

Weighted on a Mettler Toledo AB204-S balance

Ground to a fine power in the Wig-L-Bug amalgamator using stainless steel liners

Sent to Popp Lab for measurement of tissue
 15 N and
 13 C using a Carlo
Erba NC2500 Elemental Analyzer interfaced to a Delta S via a
ThermoFinnigan ConFloll (mass spectrometer) iv. Data analysis techniques: ANOVA v. Contact information for companies used to purchase items unique to your project
(if applicable): various companies sell Wig-L-Bug amalgamators – use a search engine to find suppliers (i.e. Goggle). The Popp Lab can be contacted at: http://www.soest.hawaii.edu/gg/isotope_biogeochem/aboutourlab.htm
E. Quantify and compare C:N:P of Avrainvillea amadelpha canopies
List individuals and organizations actually performing the work
F. Thomas and K.

Peyton designed the experiment; K. Peyton and K. Groom did the diving; K. Peyton collected samples; K. Peyton and L. Valentino processed samples; Popp Lab analyzed the samples ii. Material list:

Balance

Drying oven (60 o C)

Wig-L-Bug amalgamator using stainless steel liners inside threaded screw vials (plastic)

Scintillation vials

Desiccator
Data collection procedures

Collected samples of A. amadelpha from each site and transport in seawater to laboratory

Because A. amadelpha was not observed at each site, a second macroalga found at all sites, Neomaris vanbosseae, was added to the study. Where both species co-occurred they were collected together.

All samples were collected in triplicate

At the lab all samples of N. vanbosseae were scraped with a glass slide to remove epiphytes

A. amadelpha was divided in three sample types o A. amadelpha - new fronds with no visible epiphytes o Whole plant A. amadelpha – all fronds with epiphytes o Whole plant A. amadelpha – holdfast

Samples were rinsed in Nanopure water to remove sediments and salts

Samples were dried to a constant weight at 60 o C

Weighted on a Mettler Toledo AB204-S balance

Ground to a fine power in the Wig-L-Bug amalgamator using stainless steel liners

Sent to Popp Lab for measurement of tissue N and C using a Carlo Erba
NC2500 Elemental Analyzer interfaced to a Delta S via a ThermoFinnigan
ConFloll (mass spectrometer)
Data analysis techniques: ANOVA
Contact information for companies used to purchase items unique to your project (if applicable
various companies sell Wig-L-Bug amalgamators – use a search engine to find suppliers (i.e. Goggle). The Popp Lab can be contacted at: http://www.soest.hawaii.edu/gg/isotope_biogeochem/aboutourlab.htm
F. Quantify and compare heavy metals accumulated by Avrainvillea amadelpha canopies i. List individuals and organizations actually performing the work: K. Peyton and K.
Groom were the divers; K. Peyton collected samples; K. Peyton processed samples;
Centre National de la Recherche Scientifique analyzed the samples. ii. Material list

Balance

Drying oven (60 o C)

15 ml conical tissue grinders (VWR)

15 ml centrifuge tubes (VWR)


Desiccator iii. Data collection procedures

At each site Neomaris vanbosseae was collected and, when found, A. amadelpha was collected

Collected samples in triplicate

Samples were transported in seawater to the laboratory

Gloves were used when handling samples

Any epiphytes were removed by scraping the thalli with a glass slide on a plastic cutting board

For A. amadelpha only new fronds were used

New fronds were cut from the holdfast using a plastic knife to avoid introducing metals

Salts and sediments were rinse from samples using filtered seawater

Labeled 15 ml tube (VWR) were weighed

Blotted wet weight of sample was recorded

Dry sample at 60 o C for 3-5 days (constant weight)

Record dry weight of the sample to calculate wet to dry weight ratio

Use a tissue grinder to homogenize samples

Cleaned tube grinder in 1 molar acid with a clean water rinse and dry between samples/sites

desiccator

Samples were shipped in sealed tubes iv. Data analysis techniques: ANOVA
Contact information for companies used to purchase items unique to your project (if applicable
Use 15 ml centrifuge tubes and tissue grinders from VWR International
(vwr.com) because they do not introduce metals into the samples
G. Organize a workshop i. List individuals and organizations actually performing the work: Kevin Foster
(USFWS), F. Thomas and K. Peyton organized the workshop with 24 participates in attendance
A. Map the areal extent of the invasive alga Avrainvillea amadelpha along the coastline from Kalaeloa a. Accomplishments: A total of 66 sites were surveyed in Kalaeloa. Fifty-two of the sites were surveyed using SCUBA within a depth range of 25 to 90 feet. Sites are situated both north and south of the Barbers Point Harbor channel and included 66 transects. Fourteen sites were surveyed in the intertidal area during spring minus tides. Although we were requested to map the distribution of the invasive alga, we took further steps to provide managers with insights on this invasion and its status at Kalaeloa by characterizing sites with that alga as well as site were the alga was rare or not observed. b. Site specific results:

Photoquad mean percent cover across all transects:
1. Coral cover - 8.57 % (

10.1 SD)
2. Turf algae – 80.1 % ( 
14.9 SD)

3. Avrainvillea amadelpha – 6.2 % ( 
9.7 SD)
4. Native macroalgae – 0.7 % ( 
1.06 SD)
5. Seagrass – 0.14 % ( 
1.07 SD)
6. Sand (uncolonized) – 3.75 % ( 
12.7 SD)
7. Other
– 0.56 % ( 
0.7 SD)

Urchin density was positively correlated to benthic contour complexity index (1
– 10 scale with decreasing numbers indicating increasing spatial relief as measure of

A standing crop of Avrainvillea amadelpha was not detected in sites with a benthic contour complexity score of < 8 (for comparison, scores of 9-10 would be characteristic of pavement or deep-water sand plains)

A standing crop of Avrainvillea amadelpha was not detected in sites with urchin densities ca.

1 urchin m -2 (all urchin species pooled)
B. Monitor the fate of open gaps created by the physical removal of invasive algal biomass: Is the physical removal of the invasive species an option? Is there an alternative approach? a. Accomplishments: Physical removal of the invasive species by hand was a management option considered because it is being done in Maunalua Bay,
Hawaii concurrent with this project. However, during the mapping exercise from the proceeding objective (A), we interpreted the distribution patterns of A. amadelpha to be potentially influenced by macro-grazers such as urchins and fish. The opportunity was undertaken to test the hypothesis that grazers were influencing the distribution of this alga. Thus grazing experiments were substituted for the initially proposed physical removal experiment. After consulting with other researchers who have worked with grazing in Hawaii, we concluded that their cage and tether designs (used in Kaneohe Bay) would not be durable enough for Kalaeloa. Therefore, after working with several prototypes, we designed experimental cages and tethers that were able to withstand the conditions when a South Pacific swell affects the site. Given that we have experienced considerable North Pacific swell events this season, which has affected Kalaeloa significantly; we deployed the experiment in early May.
We completed one preliminary experiment and two independent experiments. b. Site specific results: Our grazing experiments demonstrated that:

Avrainvillea amadelpha is palatable to herbivores around Kalaeloa

Herbivores likely contribute to the very low standing crop of A. amadelpha on the many reefs around Kalaeloa

At the grazing rates measured during the experiment at the coral reef site an A. amadelpha plant weighting ca. 30 grams would be consumed in < 6 days

In contrast, grazing rates did not exceed growth rates at the donor site in the 68-day experiment

A standing crop of A. amadelpha can only develop at sites where grazing rates are low
C. Quantify and compare water quality inside and outside Avrainvillea amadelpha canopies a. Accomplishments: Only two sites were found with Avrainvillea amadelpha canopies extensive enough to measure water quality as planned. In a sense, this is good news, since it is an invasive species. As a result we modified the

original work plan and reallocated the resources saved by canceling further work on this objective. Resources were reallocated as follows: 1) added a second species, Neomaris (a ubiquitous, native species), to objectives D, E and F; 2) expanded the types of samples processed for Avrainvillea amadelpha in objectives D and E from one to three types; and 3) include intertidal collections of both algae in objectives D, E and F. b. Site specific results: Only two sites were found with extensive canopies.
D. Quantify stable isotopes accumulated by Avrainvillea amadelpha canopies a. Accomplishments:
 15 N and
 13 C values were determined for two species A. amadelpha or Neomaris from a total of 56 sites (not all sites had both species). b. Site specific results:
  15 N signature for A. amadelpha samples ranged from 0 to 4 0 /
00 and did not indicate a strong anthropogenically derived nitrogen source
  15 N signature for Neomaris samples ranged from 0.8 to 4.5 0 /
00
and did not indicate a strong anthropogenically derived nitrogen source
E. Quantify and compare C:N:P of Avrainvillea amadelpha canopies a. Accomplishments: Samples were collected from 56 sites at Kalaeloa. b. Site specific results:

Avrainvillea amadelpha new fronds had significantly higher percentages of nitrogen and carbon than the holdfast of this species.
A. amadelpha fronds with epiphytes were not significantly different from new fronds and from holdfasts

Neomaris had significantly lower values for both nitrogen and carbon than portion of A. amadelpha (new fronds, fronds with epiphytes, or holdfasts)
F. Quantify and compare heavy metals accumulated by Avrainvillea amadelpha canopies a. Accomplishments: Samples collected and analyzed from 56 sites at Kalaeloa and included a range of water depths. b. Site specific results: Results are still being interpreted at the time of this report.
We are working with our collaborators in France to complete this objective.
G. Organize a workshop for managers to discuss the results and next steps a. Accomplishments: Workshop was hosted at the USFWS office, Honolulu on 1
September 2010. Twenty-four participates from 9 organizations (USFWS;
University of Hawaii; Land Based Sources of Pollution Local Action Strategy
Working Group; Malama Maunalua (a local NGO); EPA; NOAA; Department of
Land and Natural Resources; Marine Corp Base-Kaneohe; Army Corp of
Engineers) attended the two-hour workshop. b. Site specific results:

Ca. 15 minute opening presentation gave the background of the site and was given by Kevin Foster

Ca. 45 minute presentation highlighted the results of this research and was given by Kim Peyton

Ca. 40 minute discussion by the group on future directions and was led by Florence Thomas, Kevin Foster and Kim Peyton
H. If significant problems developed which resulted in less than satisfactory or

negative results, they should be discussed.
We are very grateful to have been selected to receive funding from the HCRI, and we were notified that the proposal we submitted was funded. However, at a later date HCRI informed us that the focus, management questions and study site were to be entirely changed. We had a very steep learning curve and a truncated time period to both adapt and complete our new research objectives. We did encounter difficulties due to limited access at this site because of seasonal weather conditions, specifically the North Pacific swell. This fieldbased project relied on all diving work (we added the intertidal sites later in the study) and there were several periods during which we could not access the site. We had planned to wrap up diving by May and focus on sample and data analysis for the last two months of funding. However weather delays forced us to collect samples until the end of the grant period. Future projects at this site should be funded such that diving can be accomplished throughout a continuous period from April to November.
I. Description of need, if any, for additional work.
This research demonstrated the importance of grazer assemblages to maintaining a low standing crop of the invasive
A. amadelpha . However, we do not know the identity of the grazers. Urchin densities were used as a part of a rapid ecological assessment of the area. However urchins served as a proxy to quantity and compare potential grazing intensities across sites.
How much urchins grazing, both as a functional group and by species, contributed to the standing crop of Avrainvillea amadelpha is not known. We predict that herbivorous fish also play an important role in structuring the macroalgae assemblages, including the invasive species of interest. Understanding the relative importance of grazers both on the level of ecological functional groups and evolutionary lineages (families, genera, species) is clearly needed.
Outputs (Dissemination of Project results) Outputs are defined as products (e.g. publications, models) or activities that lead to outcomes (changes in user knowledge or action).
New fundamental or applied knowledge: a. Avrainvillea amadelpha is a palatable algae to macro-herbivores (fish and/or urchins) in Hawaii b. In areas with assemblages of herbivores, a standing crop of A. amadelpha was not observed
Urchin density was positively associated with increasing bottom contour (reef structures)
Standing crop of A. amadelpha was negatively associated with increasing bottom contour (reef structures)
Workshops: Workshop was held at USFWS office in Honolulu on 1 September 2010 with 24 participates
Presentations: HCRI meetings on 2 October 2009 and 5 February 2010
Partnerships established with agencies or organizations: USFWS; DLNR
A. New fundamental or applied knowledge i. Herbivore no take areas (fishing limits) could be used to control the invasion of A. amadelpha ii. Herbivore replenishment projects could be applied to controlling this invasive species iii. Management of reef structure is important to maintain herbivore densities

New or improved methods or technology
Grazing experiment improved on cage and tether designs currently in use in Hawaii. iv. Any action that reduces reef structure (i.e. ship grounding, ship channel dredging) should account for the impacts on grazers densities and macroalgae standing crop

Improved water quality

Improved management of invasive species

Improved management of coral reefs
A. Map the areal extent of the invasive alga Avrainvillea amadelpha along the coastline from Kalaeloa a. Describe the extent to which the objective was attained, including: i. Was the objective attained? How? The mapping objective was completed. GPS points of the dives along with the status of the invasive species are in a spreadsheet that will be shared with USFWS. ii. Were modifications made to objective? If so, explain. Yes, we modified this objective to include the question: Are there any patterns associated with Avrainvillea amadelpha distribution at Kalaeloa? To achieve this we used a rapid ecological assessment approach to characterize each site. From these data we determined that grazers could play an important role in structuring the standing crop of A. amadelpha . Objective (B) was modified as a result. iii. If significant problems developed, resulting in less than satisfactory or negative results, discuss. Nothing significant. iv. Description of need, if any, for additional work. Continued monitoring of the site is recommended. a. What performance measures are used to evaluate how well the project met the stated objective? The diving surveys were completed and a database was produced within the time period of the funding and the results were presented to managers in a workshop with follow up discussions.
B. Monitor the fate of open gaps created by the physical removal of invasive algal biomass b. Describe the extent to which the objective was attained, including: i. Was the objective attained? This objective was changed from a manual removal experiment to grazing experiments. ii. Were modifications made to objective? If so, explain. This objective was completely modified because the results in Objective A indicated that grazers may play important roles in limiting the standing crop of A. amadelpha . Controlled experiments were needed to test this hypothesis.
Herbivores are an important management tool and the results of this modified object underscore this critical management option. iii. If significant problems developed, resulting in less than satisfactory or negative results, discuss. None iv. Description of need, if any, for additional work. How much urchins grazing, both as a functional group and by species, contributed to the

standing crop of Avrainvillea amadelpha is not known. We predict that herbivorous fish also play an important role in structuring the macroalgae assemblages, including the invasive species of interest. Understanding the relative importance of grazers both on the level of ecological functional groups and evolutionary lineages (families, genera, species) is clearly needed c. What performance measures are used to evaluate how well the project met the stated objective? The experiments were completed within the time period of the funding and the results were presented to managers in a workshop with follow up discussions.
C. Quantify and compare water quality inside and outside Avrainvillea amadelpha canopies a. Describe the extent to which the objective was attained, including: i. Was the objective attained? How? If not, why? This objective could not be completed, as there was a lack of adequate sites with extensive A. amadelpha canopies to measure. The importance of grazers described in
Objectives A and B explain the lack of invasive species canopy. ii. Were modifications made to objective? If so, explain. We modified the original plan and used the resources saved by dropping this objective from the research plan and instead carrying out the following: 1) added a second species, Neomaris , important because it is both ubiquitous and a native species, to objectives D, E and F; 2) expanded the types of samples processed for Avrainvillea amadelpha in objectives D and E from one type to three types; and 3) included intertidal collections of both A. amadelpha and
Neomaris in objectives D, E and F. iii. If significant problems developed, resulting in less than satisfactory or negative results, discuss. The sites lacked an extensive canopy to measure; not really a problem because this is an invasive species. iv. Description of need, if any, for additional work. b. What performance measures are used to evaluate how well the project met the stated objective? This objective cannot be measured.
D. Quantify stable isotopes accumulated by Avrainvillea amadelpha canopies a. Describe the extent to which the objective was attained, including: i. Was the objective attained? How? This objective was attained because samples were collected and processed. ii. Were modifications made to objective? If so, explain. Yes, we added to this objective: 1) a second species, Neomaris , important because it is both ubiquitous and a native species; 2) expanded the types of samples processed for Avrainvillea amadelpha in objectives D and E from one type to three types; and 3) included intertidal collections of both A. amadelpha and
Neomaris in objectives D, E and F iii. If significant problems developed, resulting in less than satisfactory or negative results, discuss. No major problems. Only two minor problems: 1) weather delays extended our sampling times latter than we planned; and 2) although we regularly sent samples to the Popp Lab, they experienced repeated instrument problems in their laboratory resulting in a longer than normal cue and thus delays. It should be said that the quality of their work was worth the all delays. The Popp Lab is in the process of upgrading their instrumentation so this problem will be less likely in the future. iv. Description of need, if any, for additional work. Stable isotopes could be

applied to examining the role of A. amadelpha in the food web at Kalaeloa. b. What performance measures are used to evaluate how well the project met the stated objective? Samples were collected and processed within the time period of the funding and the results were presented to managers in a workshop with follow up discussions
E. Quantify and compare C:N:P of Avrainvillea amadelpha canopies c. Describe the extent to which the objective was attained, including: v. Was the objective attained? How? This objective was attained because samples were collected and processed. vi. Were modifications made to objective? If so, explain. Yes, we added to this objective: 1) a second species, Neomaris , important because it is both ubiquitous and a native species; 2) expanded the types of samples processed for Avrainvillea amadelpha in objectives D and E from one type to three types; and 3) included intertidal collections of both A. amadelpha and
Neomaris in objectives D, E and F vii. If significant problems developed, resulting in less than satisfactory or negative results, discuss. No major problems. Only two minor problems: 1) weather delays extended our sampling times latter than we planned; and 2) although we regularly sent samples to the Popp Lab, they experienced repeated instrument problems in their laboratory resulting in a longer than normal cue and thus delays. It should be said that the quality of their work was worth the all delays. The Popp Lab is in the process of upgrading their instrumentation so this problem will be less likely in the future. viii. Description of need, if any, for additional work. d. What performance measures are used to evaluate how well the project met the stated objective? Samples were collected and processed within the time period of the funding and the results were presented to managers in a workshop with follow up discussions
F. Quantify and compare heavy metals accumulated by Avrainvillea amadelpha canopies e. Describe the extent to which the objective was attained, including: ix. Was the objective attained? How? This objective was attained because samples were collected and processed. x. Were modifications made to objective? If so, explain. Yes, we added to this objective: 1) a second species, Neomaris , important because it is both ubiquitous and a native species; 2) expanded the types of samples processed for Avrainvillea amadelpha from one type to three types; and 3) included intertidal collections of both A. amadelpha and Neomaris xi. If significant problems developed, resulting in less than satisfactory or negative results, discuss. No major problems. Only a minor problem as weather delays extended our sampling times latter than we planned resulting in delays in shipping our sample to our colleague in France. We are still in the process of interpreting the results. xii. Description of need, if any, for additional work. f. What performance measures are used to evaluate how well the project met the stated objective? Samples were collected and processed within the time period of the funding.
G. Organize a workshop for managers to discuss the results and next steps

a. Describe the extent to which the objective was attained, including: i. Was the objective attained? How? The workshop was held on 1 September
2010 at the USFWS office. ii. Were modifications made to objective? None were needed. iii. If significant problems developed, resulting in less than satisfactory or negative results, discuss. None iv. Description of need, if any, for additional work. b. What performance measures are used to evaluate how well the project met the stated objective? The number of attendees representing a diverse group of government agencies, university researches and a NGO.