2011 OLDHAM POND ALGAE MANAGEMENT YEAR … Oldham Pond Complete Y… · 2011 Oldham Pond Algae...

2011 OLDHAM POND ALGAE MANAGEMENT YEAR-END REPORT

Prepared For:

Town of Pembroke 100 Center Street

Pembroke Watershed Association P.O. Box 368

Prepared By:

2011 Oldham Pond Algae Management Year-End Report

INTRODUCTION Oldham Pond has long supported periodic microscopic algae blooms and despite improvements made within the watershed to reduce nutrient transport the pond, the frequency and duration of the bloom conditions has become more severe in recent years. In response to these deteriorating conditions and recent public health advisories concerning toxic blue-green algae the Pembroke Watershed Association and the Town of Pembroke took steps to implement an in-pond algae control program in 2009. Due to the presence of rare mussel species in the pond the originally proposed control program (periodic copper sulfate treatment) had to be modified to address the Natural Heritage and Endangered Species Program’s (NHESP) concerns over possible impacts to the mussels. The program that was ultimately approved with specific monitoring conditions was targeted treatment with Phycomycin (sodium carbonate peroxyhyrdrate) algaecide. With all the required permits/regulatory approvals the treatment program was implemented in the summer of 2011. The following report provides a brief outline of the tasks that were performed during the course of the management program along with a discussion of the monitoring results and future management recommendations. Please accept the following as our 2011 Year–End Report for the algae management work performed at Oldham Pond. ALGAE MANAGEMENT PROGRAM METHODS Algae Monitoring Surface grab samples were collected by trained Pembroke Watershed Association (PWA) volunteers from three separate locations within the pond on a weekly basis from May 25 until August 24 (39 total samples). Each sample was preserved with gluteraldehyde and examined by Dr. Ken Wagner of Water Resource Services, Inc. for species identification and cell count by enumeration (see Attachment A – 2011 Oldham Pond Algae Control Review). The data was used to track changes in the species composition of the algal assemblage as well as species and overall cell abundance. With this information we were able to 1) identify the onset of logarithmic growth within target blue-green algae populations so that algaecide treatment could be appropriately timed and 2) identify the specific area of the pond where elevated or rapid growth first occurs so that partial pond treatment can be employed to maximize treatment efficacy and reduce potential for non-target impacts. In addition to these weekly algae samples single sample was collected on July 6th for analysis of algae toxins. Water quality MonitoringPWA volunteers again performed periodic water quality sampling at the three established algae collection sites during the course of the algae management program. The parameters that were tested included turbidity, pH, alkalinity, temperature, dissolved oxygen, and Secchi depth. In addition to this routine water quality monitoring performed by the Association, Aquatic Control staff also collected Secchi depth readings and temperature and dissolved oxygen profiles immediately prior to the Phycomycin treatments performed.

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The pH was also monitored within the selected Phycomycin treatment area throughout the application process (pre, mid, and post-treatment). Algae Control/Treatment The need for treatment was predicated upon the algae sampling/monitoring data. The Phycomycin treatment thresholds that were established for treatment were as follows; when the total blue-green algae count exceeds 10,000 cells/ml or more than doubles from the previous sample count; or when the overall cell density increases to within 30,000-35,000 cells/ml or more than doubles from the previous sample count. Once the treatment thresholds were met a treatment area of not more than 1/3 of the pond area (~65 acres) was established based on the individual sampling results. As directed by the Algal Challenge Test results (see Attachment B) a Phycomycin dose of approximately 60lbs/ac-ft. (~22 mg/l) was targeted within the top two feet of the treatment area (~120 lbs./acre). The algaecide was applied using a shallow draft 20 ft. aluminum spray boat equipped with GPS to aid in the even application of the product over the designated treatment area. The Phycomycin was applied using a calibrated eductor system that delivers the product granule using a pressurized stream of water. This treatment methodology aided in the rapid dissolution of the Phycomycin as it allows for the granule to begin dissolving in the system onboard the spray boat before being administered to the pond water. Mussel Monitoring Similar to the mussel survey work that was conducted during the permitting process, the NHESP project conditions required that a qualified mussel biologist survey state listed mussels in and adjacent to identified treatment areas. Areas supporting state listed mussels were marked to allow the mussels to be revisited following Phycomycin treatments. This monitoring work was conducted by Biodrawversity, LLC, which is the same firm that performed the preliminary mussel survey performed in 2009. A more detailed discussion of the mussel monitoring methods and results can be found in Attachment C. 2011 PHYCOMYCIN TREATMENT PROGRAM RESULTS/DISCUSSION The management program was initiated with the first round of algae samples collected in late May. The following sections provide a chronology of the various program tasks as well as a brief summary of the results of the sampling and monitoring components. 2011 Management Program Chronology Received NHESP and local Conservation Commission approvals ................... 11/10/10 Received MA DEP License to Apply Chemicals ............................................... 6/3/11 Algae sample collection dates........................................................................... 5/25; 6/2; 6/9; 6/15; 6/19; 6/28; 7/6;

7/15; 7/20; 7/27; 8/1; 8/10; 8/24 Water quality sampling dates ............................................................................ 6/19; 7/6; 7/20; 7/21; 8/3; 8/19;

8/24 Algae toxin sample collection............................................................................ 7/6/11 Phycomycin treatments..................................................................................... 6/15/11; 7/15/11; 8/18/11 Mussel surveys ................................................................................................. 6/4/11; 6/21/11; 9/7/11

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Algae Monitoring Blue-green algae growth accelerated quickly in the early part of the growing season, where the algae density went from zero to an average of 75,000 cells/ml in the first three weeks of the program. This early season blue-green algae assemblage was dominated by the filamentous nitrogen fixers (Anabaena and Aphanizomenon) and the unicellular colonial species (Woronichinia and Microcystis). The initial Phycomycin treatment provided a significant (~70%) reduction in blue-green algae density. The cell densities continued to rise until additional treatment was required to maintain and further reduce the blue-green algae growth. The graph below shows the total blue-green algae cell density over the course of the season. More detailed information is provided in the Water Resource Services, Inc. Report provided in Attachment A.

OLDHAM POND CYANOPHYTA CELL DENSITY

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10000

20000

30000

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5/25

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6/1/

2011

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6/22

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6/29

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7/13

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Sample Collection Dates

Cya

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cells

/ml

Site 1Site 2Site 3Average

6/15/11 Phycomycin 7/15/11 Phycomycin Treatment Treatment

8/18/11 Phycomycin Treatment

Water Quality Monitoring Temperature and dissolved oxygen profiles collected throughout the season showed fairly typical values for a shallow algae infested lake. In many instances during the course of the summer dissolved oxygen levels were at or above saturation in the upper level of the water column, which is not uncommon under dense phytoplankton growth. The profiles also showed a decline in dissolved oxygen concentrations just off the lake bottom, which likely attributable to normal Biochemical Oxygen Demand (BOD) resulting from the breakdown of organic material. The Phycomycin treatments did not appear to result in any significant alteration of these typical temperature/dissolved oxygen profiles. In fact some of the later season data seems to suggest that dissolved oxygen levels were more greatly impacted during periods of non-treatment when algae densities were at there peak. The temperature and dissolved oxygen data are provided in Attachment D. In addition to the temperature and dissolved oxygen data, samples were periodically collected throughout the season to analyze pH, total alkalinity, and turbidity. The data shows an increase in alkalinity over the course of the summer and a drop in pH to about neutral at the end of the season. Abundant algae growth can influence both pH and total alkalinity, so these fluctuations are in part the result of changing algae densities and species composition. Turbidity values were high, as turbidity values rarely rise above 5-10 NTU in unpolluted waterbodies. These elevated levels are significantly influenced by the high density of microscopic algae cells. The sampling results are provided in the following table.

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Table 1 – Oldham Pond 2011 Water Sampling Results Date Site pH (S.U.) Alkalinity (mg/l) Turbidity (NTU)

1 9.4 19.2 29 2 9.29 19.3 24 6/19/11 3 8.45 18.3 23 1 8.75 19.1 13 2 8.86 19.5 19 7/6/11 3 9.15 17.4 24 1 6.8 20.2 17 2 6.8 22.7 19 7/20/11 3 6.8 20.4 18 1 7.2 25.1 8.7 2 7.0 25.9 13 8/3/11 3 7.2 25.7 13 1 7.2 27.3 13 2 7.2 28.7 13 8/24/11 3 7.0 28.6 14

Pre & Post-Treatment Mussel Surveys As required by NHESP, comprehensive mussel surveys were conducted to evaluate the possible impacts of the Phycomycin treatment program on the rare mussel population. Although some limited mussel mortality and stress was observed within the treatment area study plots, it was determined that is was not significantly different to the stresses and mortality that was observed in the control plots. It was therefore the opinion of Biodrawversity, LLC that the differences in mussel behavior and densities pre and post-treatment were more likely associated with natural habitat variations rather than any response to the Phycomycin treatment. Overall the mussel monitoring yielded very little evidence that would suggest that the Phycomycin treatments performed at Oldham Pond had any short-term adverse effects on the resident freshwater mussel population. The full Biodrawversity, LLC mussel monitoring report is provided in Attachment C. SUMMARY The phytoplankton growth experienced at Oldham pond in 2011 is consistent with previous years and proved to be difficult to manage effectively. Most available literature suggests that the efficacy of peroxide-based algaecides like Phycomycin is inversely proportionate to blue-green algae densities; therefore, the incredibly rapid growth experienced at the start of the season (0-75,000 cells/ml) made achieving a high level of control more difficult. Despite initiating the first Phycomycin treatment at a blue-green algae density higher than what would be considered optimal, a significant level of control was achieved. Overall the algae monitoring data does suggest that the treatments provided a positive reduction in blue-green algae density over the course of the season, which ultimately led to the pond being reopened to recreational use. Based on the data that was collected, our recent experience with this new product, and consultation with other experts in the field we are in the process of refining the algae management program for 2012. We are currently exploring alternative and expanded Phycomycin treatment options as well as the possible inclusion of nutrient precipitation strategies. Given the relatively severe blue-green algae growth conditions that have been occurring in Oldham Pond, we feel that some form of continued algae management along with continued efforts to reduce the external nutrient load are important to not only protect recreational safety, but the unique habitat of the resident freshwater mussel population.

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APPENDIX A

2011 Oldham Pond Algae Control Review

Water Resource Services, Inc.

2011 OLDHAM POND

ALGAL CONTROL REVIEW

PREPARED FOR AQUATIC CONTROL TECHNOLOGY

BY WATER RESOURCE SERVICES, INC.

JANUARY 2012

Contents

Introduction ..................................................................................................................................................1

Project Approach ..........................................................................................................................................1

Results...........................................................................................................................................................3

Discussion ...................................................................................................................................................10

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Introduction

Oldham Pond, in Pembroke, Massachusetts, has experienced an increasing frequency of algal blooms involving cyanobacteria (blue‐green algae). Given the presence of endangered mollusks, concern was expressed over the potential use of copper based algaecides to alleviate bloom conditions. It was decided that a peroxide based algaecide would be more appropriate. Although the use of peroxide based algaecides has been allowed in other states for many years, approval for this class of algaecide was only recently granted in Massachusetts, and this was the first permitted use of a modern peroxide based algaecide in Massachusetts.

Aquatic Control Technology of Sutton, Massachusetts performed the treatment in response to increasing cyanobacteria cell counts in summer of 2011. This report documents the treatment and its results with regard to algae in Oldham Pond.

Project Approach Oldham Pond covers approximately 220 acres. Two areas of up to about 65 acres each were targeted for possible treatment with Phycomycin, a peroxide based algaecide distributed by Applied Biochemists of Wisconsin. Three monitoring stations were established, one in each targeted treatment area and one in the untreated area (Figure 1). Surface samples were collected roughly weekly by local volunteers between late May and late August, preserved with gluteraldehyde for algal analysis, and sent to WRS in Wilbraham, MA. Algal analysis was performed by WRS within a week, keeping the monitoring process close to current. Algae were concentrated then quantitatively examined under phase contrast microscopy to determine the overall and relative abundance of algal types. Cell concentrations were calculated from the counts and biomasses were estimated based on cell dimensions and an assumption of a specific gravity of 1.000. Some additional samples were collected and analyzed by the MA DEP and yet others were collected and sent to GreenwaterLaboratiories of Palatka, Florida for additional analyses. Phycomycin was applied when cyanobacteria exhibited a sharp increase, with the area near the town beach and the northwest portion of the lake treated on June 15, 2011 (Figure 1). Phycomycin was added at a dose of 120 lbs per acre, amounting to 60 lbs per acre for each of the upper two feet of water, or a concentration of approximately 22 mg/L for those two acre‐feet. Based on monitored algal levels, an additional treatment involving the same dose was conducted in the area that included the town swimming area on July 15, 2011, and yet another treatment was performed in this area on August 18, 2011 (Figure 1). Between 7800 and 8000 lbs of phycomycin was applied in each treatment.

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Figure 1.Treatment areas and monitoring sites in Oldham Pond in 2011.

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Results

Algae observed in Oldham Pond during the summer of 2011 included 60 genera from 15 identified functional groups within 7 major algal groups (usually recognized as phyla) (Table 1). Of the identified genera, only 12 genera comprised more than 1% of either cell counts or estimated biomass, and over half of these were cyanobacteria: Aulacoseira (diatom), Micractinium (chlorococcalean green), Staurastrum (desmid green), Trachelomonas (euglenoid), Ceratium(dinoflagellate), Aphanocapsa, Microcystis, Woronichinia (coccoid cyanobacteria), Anabaena, Aphanizomenon (N‐fixing filamentous cyanobacteria), Planktolyngbya and Pseudanabaena(filamentous cyanobacteria).

In terms of cell counts, diatoms (Bacillariophyta) represent 2.6% of all algae counted, while green algae (Chlorophyta) represent 3.3% (Table 2). Golden algae (Chrysophyta), cryptomonads (Cryptophyta), euglenoids (Euglenophyta) and dinoflagellates (Pyrrhophyta) each provide no more than 0.2% of the total count. This leaves cyanobacteria accounting for the majority of algal cells (93.7%), with coccoid forms providing 54.4%, filamentous nitrogen fixers contributing 20.6%, and filamentous non‐nitrogen fixers comprising 18.7% of the total cell count.

Based on biomass as calculated from cell counts and average cell dimensions, cyanobacteria again account for the majority of algal biomass (87.2%), with coccoid forms providing 18.3%, filamentous nitrogen fixers contributing 71.1%, and filamentous non‐nitrogen fixers comprising 0.8% of the total biomass(Table 2).Diatoms (Bacillariophyta) represent 3.9% of all algal biomass, while green algae (Chlorophyta) represent 6.3%. Golden algae (Chrysophyta) at 0.04% andcryptomonads (Cryptophyta) at 0.11% are very minor biomass components. Euglenoids (Euglenophyta) contribute 1.1% of biomass and dinoflagellates (Pyrrhophyta) provide 1.3% of the algal biomass. While the shift from cell counts among major algal groups is not extreme, differences in average cell size to make a difference. Within the cyanobacteria, the cells of Anabaena are much larger than other cyanobacteria in Oldham Pond, leading to its dominance of total biomass.

Cell counts (Figure 2) and biomass estimates (Figure 3) reveal the pattern of algal abundance, both overall and among major algal groups. There are substantial shifts in composition among sampling dates that cause appreciable differences in the patterns of cell counts and biomass, but there is relatively less variability among the three sampled stations on any given date (Figures 2 and 3). The red line on Figure 2 marks the 70,000 cells/mL threshold used by the Commonwealth of Massachusetts to signal a toxicity warning when cyanobacteria are dominant. The red line on Figure 3 at 10,000 ug/Lis an unofficial but generally recognized threshold for very high algal biomass in recreational lakes, while the green line at 2500 ug/L indicates a biomass that normally results in visual impairment of water clarity.

The cell count threshold was not often exceeded based on the WRS analysis, but it has been reported that both the MA DEP and Greenwater Laboratory analyses indicated higher counts and more frequent exceedences of the cell count threshold. Direct comparison with one set of Greenwater Laboratory results revealed the difference to be almost entirely due to higher Greenwater counts of the coccoid

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Table 1.Distribution of cells and biomass among genera.

TAXON Cells/mL ug/L

Filamentous ChlorophytesOedogonium 0.01% 0.03%

DesmidsClosterium 0.07% 1.22%Cosmarium 0.01% 0.03%Euastrum 0.01% 0.03%Staurastrum 0.34% 1.21%Staurodesmus 0.01% 0.02%

CHRYSOPHYTAFlagellated Classic ChrysophytesChromulina 0.00% 0.00%Mallomonas 0.01% 0.03%Uroglena 0.02% 0.01%

Non‐Motile Classic Chrysophytes

Haptophytes

Tribophytes/Eustigmatophytes

Raphidophytes

CRYPTOPHYTACryptomonas 0.10% 0.11%

CYANOPHYTAUnicellular and Colonial FormsAphanocapsa 5.47% 0.24%Chroococcus 0.01% 0.00%Microcystis 21.23% 13.81%Woronichinia 27.71% 1.23%

Filamentous Nitrogen FixersAnabaena 12.86% 66.64%Aphanizomenon 7.77% 4.48%

Filamentous Non‐Nitrogen FixersPlanktolyngbya 6.04% 0.27%Pseudanabaena 12.62% 0.56%

EUGLENOPHYTAEuglena 0.00% 0.01%Phacus 0.00% 0.00%Trachelomonas 0.19% 1.12%

PYRRHOPHYTACeratium 0.02% 1.19%Peridinium 0.01% 0.11%

% of Total

TAXON Cells/mL ug/L

BACILLARIOPHYTACentric DiatomsAcanthoceras 0.01% 0.04%Aulacoseira 2.37% 3.16%Urosolenia 0.00% 0.02%

Araphid Pennate DiatomsAsterionella 0.04% 0.02%Fragilaria/related taxa 0.03% 0.04%Synedra 0.10% 0.46%Tabellaria 0.00% 0.01%

Monoraphid Pennate DiatomsAchnanthidium/related taxa 0.00% 0.00%

Biraphid Pennate DiatomsNavicula/related taxa 0.02% 0.05%Nitzschia 0.01% 0.05%Pinnularia 0.00% 0.07%

CHLOROPHYTAFlagellated ChlorophytesChlamydomonas 0.01% 0.01%Coccomonas 0.00% 0.00%Eudorina 0.20% 0.35%

Coccoid/Colonial ChlorophytesActinastrum 0.06% 0.02%Ankistrodesmus 0.15% 0.07%Botryococcus 0.06% 0.05%Closteriopsis 0.02% 0.04%Coelastrum 0.34% 0.30%Dictyosphaerium 0.28% 0.12%Elakatothrix 0.03% 0.01%Golenkinia 0.02% 0.02%Kirchneriella 0.01% 0.00%Lagerheimia 0.00% 0.00%Micractinium 0.09% 1.17%Oocystis 0.07% 0.12%Paulschulzia 0.02% 0.04%Pediastrum 0.31% 0.25%Quadrigula 0.00% 0.00%Scenedesmus 0.30% 0.13%Schroederia 0.04% 0.43%Selenastrum 0.00% 0.00%Sorastrum 0.04% 0.04%Sphaerocystis 0.81% 0.52%Tetraedron 0.00% 0.01%Tetrastrum 0.00% 0.00%Treubaria 0.00% 0.01%

% of Total

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Table 2.Summary of phytoplankton cells and biomass per liter by selected groups.

Cells/mL ug/LDENSITY (CELLS/ML) SUMMARYBACILLARIOPHYTA 2.59% 3.93% Centric Diatoms 2.38% 3.22% Araphid Pennate Diatoms 0.17% 0.54% Monoraphid Pennate Diatoms 0.00% 0.00% Biraphid Pennate Diatoms 0.04% 0.17%CHLOROPHYTA 3.33% 6.26% Flagellated Chlorophytes 0.21% 0.36% Coccoid/Colonial Chlorophytes 2.68% 3.37% Filamentous Chlorophytes 0.01% 0.03% Desmids 0.43% 2.50%CHRYSOPHYTA 0.04% 0.04% Flagellated Classic Chrysophytes 0.04% 0.04% Non‐Motile Classic Chrysophytes 0.00% 0.00% Haptophytes 0.00% 0.00% Tribophytes/Eustigmatophytes 0.00% 0.00% Raphidophytes 0.00% 0.00%CRYPTOPHYTA 0.10% 0.11%CYANOPHYTA 93.72% 87.23% Unicellular and Colonial Forms 54.42% 15.28% Filamentous Nitrogen Fixers 20.63% 71.12% Filamentous Non‐Nitrogen Fixers 18.67% 0.83%EUGLENOPHYTA 0.20% 1.12%PYRRHOPHYTA 0.03% 1.30%

% of Total

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Figure 2.Phytoplankton cell counts for Oldham Pond in 2011.

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Figure 3.Phytoplankton biomass for Oldham Pond in 2011.

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cyanobacteriumWoronichinia. These higher counts are attributed to the use of Lugol’s solution as a preservative, causing dissociation of colonies; this makes counting easier and usually more accurate, but also usually results in less of the sample being counted and higher multiplication factors for conversion to cells/mL. MA DEP counts were reportedly intermediate to those of WRS and Greenwater Laboratories. Taxonomic identifications and the pattern of algal abundance were very similar among sample results, however, indicating that methods affected mainly total quantity estimation, not the qualitative aspects or relative abundance of algal types.

The pattern of biomass suggests that water clarity will be impaired for nearly the entire monitoring period, with very high biomasses observed in June and to a lesser extent in July. While it is reasonable to state that cyanobacterial abundance was high enough to impair water quality nearly the whole summer, it is not clear that cell counts are a reliable indicator of problematic biomass. This is a function of highly variable cell size, with most cyanobacterial cells at the small end of the algal cell scale and requiring very high cell counts to produce high biomass. However, even within the cyanobacteria, the range of cell sizes is large, and high cell counts for Woronichinia provide lower biomass than much lower cell counts of Anabaena.

The general pattern of cell counts (Figure 2) includes moderate levels of green algae in May giving way to elevated levels of the cyanobacteria Woronichinia, Microcystis, Anabaena andAphanizomenonin June. After the June 15th treatment, cyanobacteria temporarily declined, but then rebounded in July, with more coccoid forms (Woronichinia and Microcystis) than filamentous nitrogen fixers (Anabaena and Aphanizomenon), based on cell counts. After the July 15th treatment Anabaena and Aphanizomenonweregreatly depressed initially and Woronichinia and Microcystisdeclined to a lesser extent but still substantially. All of these rebounded in late July and early August, but moreso for the coccoid forms, and with a concurrent rise in the non‐nitrogen fixing filamentous cyanobacteriaPlanktolyngbyaandPseudanabaena. As August progressed, Woronichinia declined noticeably while Microcystis, Anabaena andAphanizomenonwere moderately stable and PlanktolyngbyaandPseudanabaenaincreased somewhat. After the August 18th treatment Woronichinia disappeared, Pseudanabaena increased, and the other cyanobacteria were moderately stable.

The pattern of algal biomass (Figure 3) is different from that for cell counts. Biomass is very high in June and vested mainly in Anabaena and Aphanizomenon. Biomass is reduced to near the very high threshold of 10,000 ug/L after the June 15th treatment, and oscillates around that level for most of the rest of the summer. After the June treatment, dominance is shared by filamentous nitrogen fixers and coccoid cyanobacteria. Despite the rise of non‐nitrogen fixing filamentous cyanobacteria later in summer, they never dominate the biomass. Fluctuations suggest some impact from subsequent treatments, and algal biomass is generally lower at stations 1 and 3 which were treated at some point vs. station 2, which was never directly treated, but the pattern of biomass levels is similar at all stations. Biomass is more evenly divided among major algal division in the final August samples, a week after the last treatment, but the least even distribution and greatest remaining cyanobacteria biomass was at station 1 within the August 18th treatment area.

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Discussion

The cyanobacteria bloom in Oldham Pond during summer of 2011 was similar to blooms encountered in many other southern New England lakes in 2011 and recent years. There appears to be justifiable concern over increasing cyanobacteria blooms from the perspective of recreational safety, including reduced clarity and possible toxin exposure, and from the perspective of ecological damage, including water quality fluctuations (especially pH and oxygen), altered energy flow (emphasizing detrital pathways) and again potential toxicity. Controlling nutrient levels in lakes appears to be highly desirable, but this is not always feasible. However, algal treatment options are limited and algal monitoring requires expertise not always available in a timely manner to facilitate the most effective treatment approach. Peroxide based algaecides have been enhanced and received wider use in recent years, and became approved for use in Massachusetts in 2011. This first application, to Oldham Pond, was monitored to provide potentially helpful data, but was not done as a research project. The Oldham Pond treatment in 2011 represents an effort to control cyanobacteria while minimizing the threat to endangered mollusks. Mollusk impacts were not reported, but are not part of this assessment.

There is evidence in the data that treatment had an impact, at least temporarily, but the pattern of both cell counts and biomass over space is fairly consistent when treatments were focused on only about a 65 acre area on any of the three treatment dates. Either the impact of treatment extended well beyond the area treated, or other factors are more important to the observed algal fluctuations. The shift in composition of the algal community is consistent with expectations based on natural factors often observed in untreated lakes (e.g., nitrogen fixers giving way to non‐nitrogen fixers), but the decrease in biomass after June is atypical. Still, algal biomass was above desirable levels throughout the lake during most of the summer, despite repeated treatments, and cyanobacteria remained dominant. Treatment may have reduced biomass and shifted the relative abundance of species, but it did not achieve conditions that would be perceived by most lake users as desirable.

With regard to toxins, we are aware of testing for only one date, July 6, 2011, and levels of microcystin, anatoxin and saxitoxin were all undetectable. Detection limits were below any recognized threshold for ecological or human impact, so despite elevated cyanobacterial cell counts and biomass, toxins represented no significant threat. However, the samples were collected three weeks after the peak counts and biomasses were detected and the first phycomycin treatment was conducted. It is possible that toxins were present earlier and that toxin production was reduced by the treatment, but we have no data that can be used to conclusively evaluate this possibility.

APPENDIX B

Responses of algae from Oldham Pond located in Pembroke, MA

to a peroxide algaecide exposure (Phycomycin® SCP)

Clemson University

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Responses of algae from Oldham Pond located in Pembroke, MA to a peroxide algaecide exposure

(Phycomycin® SCP)

Prepared by:

West M. Bishop, M.S., Graduate Research Assistant, Clemson University [email protected]

Ben E. Willis, B.S., Graduate Research Assistant, Clemson University

[email protected]

John H. Rodgers, Jr. Ph.D., Professor, Clemson University [email protected]

Department of Forestry and Natural Resources Clemson University, 261 Lehotsky Hall

Clemson, SC 29634

P: 864.656.0886 F: 864.656.3304

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Recommendation for treatment of algae sampled from Oldham Pond located in Pembroke, MA:

For treatment of algae sampled from Oldham Pond located in Pembroke, MA we recommend the following:

1. Application of 22.14 mg Phycomycin® SCP/ L (60 pounds/ acre-ft)

This recommendation is based on chlorophyll a concentrations, cell densities, total microcystin, visual observations and a partial application label rate of Phycomycin® SCP. Note: with a decline in cyanobacteria in effective treatments there was an increase in other algae and chlorophyll a. Subsequent applications may be necessary. Follow all algaecide label directions. Sample From: Oldham Pond: Pembroke, MA 02359 Company / Agency: Aquatic Control Technology, Inc. Site Contact Person: Keith Gazaille (Senior Biologist) Phone: (508) 865-1000 Applied Biochemists Contact: Bill Ratajczyk Phone: (608) 524-4014 Date Received: August 24, 2010 Experimental Period: August 26 – August 30, 2010 Test Completed: Responses of algae, water characteristics, and microcystin following an algaecide exposure Formulations Tested:

Phycomycin® SCP (4 day exposure)

Types of Algae and Initial Densities: Cyanophyta (Blue-green algae) Pseudanabaena sp. 2.5 x 104 cells/ mL Aphanizomenon sp. 2.1 x 104 cells/ mL Anabaena sp. 3.7 x 103 cells/ mL Microcystis sp. < 1,000 cells/ mL

Initial Chlorophyll a Concentration: 21.9µg chlorophyll a/ L

Initial Total Microcystin Concentration: 0.42ppb total microcystin

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Initial water characteristic measurements:

Oldham PondpH (SU) 7.30

Dissolved Oxygen (mg O2 / L) 8.60 Alkalinity (mg/ L as CaCO3) 28 Hardness (mg/ L as CaCO3) 36

Conductivity (µS/ cm) 240.6 Turbidity (NTU) 9.4

Temperature (°C) 24.8

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Appendix

1. Appendix A……………...Responses of algae sampled from Oldham Pond located in Pembroke, MA to exposures of Phycomycin® SCP.

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Appendix A

Responses of algae sampled from Oldham Pond located in Pembroke, MA to exposures of Phycomycin®.

A Site-Specific Algaecide Study

Sample From: Oldham Pond: Pembroke, MA 02359 Company / Agency: Aquatic Control Technology, Inc. Site Contact Person: Keith Gazaille (Senior Biologist) Phone: (508) 865-1000 Applied Biochemists Contact: Bill Ratajczyk Phone: (608) 524-4014 Date Received: August 24, 2010 Experimental Period: August 26 – August 30, 2010 Test Completed: Responses of algae, water characteristics, and microcystin following an algaecide exposure Formulations Tested:

Phycomycin® SCP (4 day exposure)

Types of Algae and Initial Densities: Cyanophyta (Blue-green algae) Pseudanabaena sp. 2.5 x 104 cells/ mL Aphanizomenon sp. 2.1 x 104 cells/ mL Anabaena sp. 3.7 x 103 cells/ mL Microcystis sp. < 1,000 cells/ mL

Initial Chlorophyll a Concentration:

21.9µg chlorophyll a/ L

Initial Total Microcystin Concentration: 0.42ppb total microcystin

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Growth Chamber Characteristics: - Maintained at 22 ± 2°C - 16-h light / 8-h dark photoperiod - Light intensity of 3077 lux

Test Specifics:

• Experimental objective was to obtain control of algae. • Algae were exposed to three concentrations in 500 mL flasks. • Experiments were initiated using 500 mL site water. • The toxicity experiment was initiated by exposing the algae to 7.38, 14.76, 22.14,

29.52 and 36.9 mg Phycomycin® SCP/ L. Three replicates of each exposure concentration, along with three replicates of untreated controls, were tested.

7

Algal Response Results (see Table 1): Table 1: Responses of algae in untreated controls and exposure concentrations of Phycomycin® SCP four days after treatment.

Phycomycin® SCP (mg/ L)

Day 4 Visual Observations

Avg. Day 4 Cyanobacteria Cell

Densities (cells/ mL)

Avg. Day 4 Other Algae Cell

Densities (cells/ mL)

Avg. Day 4 Chlorophyll-a

(µg/ L)

Avg. Day 4 Total

Microcystin (ppb)

Untreated Control Algae on bottom,

suspended & floating, green

Aphanizomenon 1.29 x 105 Microcystis 1.7 x 104

Pseudanabaena 1.5 x 104 Anabaena < 1 x 103

Scenedesmus < 1 x 103 Ankistrodesmus < 1 x 103

Ulothrix < 1 x 103 Other < 1 x 103

23.2 0.76

7.38 (20 lbs/ acre-ft)

Algae on bottom, suspended &

floating, green

Aphanizomenon 2.87 x 104 Microcystis < 1 x 103


Scenedesmus < 1 x 103 Ankistrodesmus 2.7 x 103

Ulothrix 8.7 x 103 Other < 1 x 103

18.7 0.72

14.76 (40 lbs/ acre-ft)

Algae on bottom & suspended, light

green

Aphanizomenon < 1 x 103 Microcystis < 1 x 103


Scenedesmus < 1 x 103 Ankistrodesmus 6.7 x 103

Ulothrix 7.0 x 103 Other 9.3 x 103

14.1 *

22.14 (60 lbs/ acre-ft)


green



Scenedesmus 7.0 x 103 Ankistrodesmus 5.0 x 103


16.6 0.41

29.52 (80 lbs/ acre-ft)


green





18.1 *

36.9 (100 lbs/ acre-ft)


green





23.1 0.37

* = No measurement taken Other = densities of other algae are combined and include the genera: Coelastrum, Pediastrum, Staurastrum, Gloeocystis

8

Exposure Water Characteristics (see Table 2): Table 2: Water characteristics in untreated controls and exposure concentrations of Phycomycin® SCP four days after treatment. Phycomycin®

SCP (mg/ L)

pH (SU) Dissolved

Oxygen (mg O2 / L)

Alkalinity (mg/ L as CaCO3)

Hardness (mg/ L as CaCO3)

Conductivity (µS/ cm)

Turbidity (NTU)

Temperature (°C)

Untreated Control 9.1 10.61 24 34 235.2 11.3 24.4

7.38 (20 lbs/ acre-ft) 9.3 11.56 28 34 253.5 6.3 25.9

14.76 (40 lbs/ acre-ft) 9.3 11.42 34 34 259.8 8.1 25.9

22.14 (60 lbs/ acre-ft) 9.3 12.20 36 32 269.5 8.2 25.9

29.52 (80 lbs/ acre-ft) 9.5 12.18 40 42 283.7 7.2 25.4

36.9 (100 lbs/ acre-ft) 9.1 12.77 44 26 292.1 6.9 24.5

9

Summary: If Phycomycin® is chosen for treatment of algae sampled from Oldham Pond located in Pembroke, MA, we recommend an application of 22.14 mg Phycomycin®/ L (60 pounds/ acre-ft) to control the cyanobacteria. Subsequent applications may be necessary. Follow all algaecide label directions.

APPENDIX C

Freshwater Mussel Monitoring for the Oldham Pond Algaecide Treatment

Biodrawversity, LLC

Freshwater Mussel Monitoring for theOldham Pond Algaecide Treatment

Prepared By:Biodrawversity LLC

433 West StreetAmherst, Massachusetts 01002

Prepared For:Aquatic Control Technology

11 John Road, Sutton, MA 01590

January 2012

biodrawversity

ecological consultingand communications

NHESP File Number 09-26729

1

BIODRAWVERSITY TECHNICAL REPORTFreshwater Mussel Monitoring for the Oldham Pond Algaecide Treatment

INTRODUCTION

Oldham Pond, a 235-acre lake in Hanson and Pembroke, Massachusetts, is on the state’s list of impaired waterbod-ies (Category 4c of the 2002 Integrated List of Waters). This listing is mainly due to the presence of invasive aquatic plants, although in recent years the pond’s water quality has deteriorated from nutrient enrichment and ex-cessive algal blooms. Oldham Pond has an average depth of 10 feet and a maximum depth of 15 feet. A small tribu-tary, several cranberry bogs, and various other wetlands are within the pond’s drainage area, and its outlet stream flows south into nearby Furnace Pond (which is higher on the state’s list of impaired waterbodies (Category 5 of the 2002 Integrated List of Waters), for organic enrichment and low dissolved oxygen). The uplands surrounding Old-ham Pond’s 2.8 miles of shoreline are heavily developed with seasonal and year-round homes. The pond typically experiences considerable recreational use during the sum-mer, but during the summer of 2011 the public was ad-vised to not use the pond because high levels of toxic blue-green algae were detected.

The poor water quality in Oldham Pond has had notable adverse effects on the lake’s biota in the past de-cade or so. In a 2009 study (Biodrawversity 2009), tens of thousands of mussel shells (of many species) were ob-served throughout the lake, suggesting mass mortality

within the last ten years, probably related to eutrophic conditions and low levels of dissolved oxygen. MassWild-life’s profile for Oldham Pond states that a mussel kill oc-curred in 1999, and it seems likely that this phenomenon may occur annually due to low oxygen stemming from or-ganic enrichment (although the magnitude of these events may vary). Mussels are still relatively common in the lake, especially the eastern elliptio (Elliptio complanata), eastern lampmussel (Lampsilis radiata), and eastern floater (Py-ganodon cataracta), suggesting that the mussel community

Oldham Pond in Hanson and Pembroke, Massachusetts, has chronic nutrient problems and turns green each year.

Eastern Pondmussel (Ligumia nasuta).

2


Two SCUBA divers worked together to set up the plots, and the procedure was the same for all plots. First, plots were placed on the lake bottom. Then the sides of each plot were fortified with small cobbles to prevent mussels from leaving the area of the plot. Next, plots were then cleared of all live mussels and shells. Finally, survey-ors placed a precise number of mussels (25) into the cen-ter 0.25m2 of each plot; these mussels consisted of ten eastern elliptio, ten eastern lampmussel, and five eastern

is persisting de spite annual algal blooms and anoxic con-ditions. However, the state-listed tidewater mucket (Lep-todea ochracea), which was once known to occur in Old-ham Pond, was not found either in 2009 or 2011, and it might not still be extant in the lake. The small population size of a second state-listed mussel, the eastern pondmus-sel (Ligumia nasuta), suggests that this species might be critically imperiled in Oldham Pond.

Given the overall deteriorating condition of Oldham Pond and the recent public health advisories for toxic al-gae, lake stewards decided to take action. Although most people agreed that control of nonpoint source pollution from the heavily developed upland landscape would pro-vide the best long-term benefits for the lake, a short-term solution of chemically treating the lake was proposed. The treatment was permitted. Among the conditions of the permit was a requirement by the Massachusetts Natural Heritage and Endangered Species Program (NHESP) to carefully monitor chemical parameters and assess the response (e.g. mortality and behavior) of mussels to the treatments.

In an attempt to control seasonal blooms of blue-green algae, Phycomycin (sodium carbonate peroxyhy-drate) was applied to the pond by Aquatic Control Tech-nology (ACT) on June 15, July 15, and August 18, 2011. Very little is known about the effects of Phycomycin treatments on non-target species of lake biota, especially freshwater mussels, but some types of chemicals used to control algae and plants are known to have adverse effects (Mattson et al. 2004).

METHODS

Pre-Treatment Experimental SetupDates: June 4-5, 2011Three survey plots were located in the northwest bay of Oldham Pond, in areas where Phycomycin treatments were planned (Figure 1, Table 1). Additionally, three con-trol plots were located east of Monument Island, away from the treatment area. The physical plots consisted of 2.25 m2 (1.5m x 1.5m) quadrat frames made of PVC pip-ing with integral weights.

Figure 1. Aerial view of Oldham Pond and nearby waterbodies. Red dots indicate approximate locations of the mussel monitoring plots in the treated areas of Oldham Pond and areas that served as a control.

The satellite image was captured in early July 2008 when lake-wide algal blooms were prevalent in Furnace Pond (note the floating algal scum along the shorelines), algal counts were also high throughout Oldham Pond but less severe than in Furnace Pond, and Great Sandy Bottom Pond exhibited the color and clarity of a more “typical” oligo-trophic pond without algal problems (image source: Google Earth).

treatment plots

control plots

Furnace Pond

Great SandyBottom Pond

Oldham Pond

N

Type Plot Latitude Longitude Depth (feet) SubstrateControl 1 42.0657400 -70.8378230 6 sand, gravel, small cobbleControl 2 42.0657550 -70.8378160 9 sand, gravel, small cobbleControl 3 42.0657670 -70.8378130 8 sand, gravel, small cobbleTreatment 1 42.0703800 -70.8407810 10 sand, cobble, organic materialTreatment 2 42.0703990 -70.8407800 8 sand, cobble, organic materialTreatment 3 42.0704020 -70.8408060 10 sand, cobble, organic material

Table 1. Location and habitat data for the control and treatment mussel monitoring plots in Oldham Pond.

3


floater. Too few live eastern pondmussels were found to include in these plots.

First Post-Treatment MonitoringDates: June 21-22, 2011In the first post-treatment monitoring, surveyors mea-sured various parameters to assess any deleterious effects to mussels seeded in the treatment and control plots. Since some of the parameters that we measured were subtle and subjective, we sought consistency by having a single expe-rienced mussel biologist (Jeffrey Cole) be responsible for recording all survey data during the first post-treatment monitoring. Parameters included:• Counts: All animals evident at the surface were

counted, and then plots were excavated to find the buried animals; these counts were recorded separately.

• Mortality: Animals were considered dead when they were unresponsive, gaping (which results from death, due to loss of function in the adductor muscles that typically pull valves shut), or showed signs of decom-position. Tightly closed mussels were considered alive.

• Embeddedness: For each mussel visible at the surface (i.e., not buried), the proportion (percent) of total shell length below the surface was estimated. Mussels lying on their sides on the surface were considered zero percent embedded, while those with just the tips of their mantle apertures showing were considered 95 percent embedded. Mussels found during excavation (i.e., buried) were considered 100 percent embedded.

• Filtering: For each mussel observed at the surface, surveyors noted whether its valves were open or closed. This was recorded as 0 (closed), 0.5 (partially open), or 1.0 (open). Later, an average filtering score was computed for all visible mussels in a given plot. A filtering score was not recorded for buried mussels.

• Responsiveness: The responsiveness of all mussels at the surface was recorded. Responsiveness was deter-mined by observing whether or not valves were open, noting the time it took a mussel to close its valves when disturbed, seeing if the mussel ejected feces or glochidia when touched, recording the time it took for a mussel to re-open its valves after being disturbed, and assessing whether or not the foot was extended. Responsiveness was recorded as 0 (not responsive), 0.5 (somewhat responsive), or 1.0 (responsive). A re-sponsive score was then computed as the average for all the mussels within a plot. Responsiveness was not measured for buried mussels.

After data were collected, the plots were removed from the lake bottom because at the time no further monitoring was anticipated.

quadrats in treated areas

Oldham Pond

Figure 2. Aerial view of Oldham Pond showing locations of the 42 4.0 m2 quadrats used in the second post-treatment monitoring. Red dots indicate quadrat locations where single eastern pondmussels were found, whereas all other quadrats are shown with a yellow dot.

quadrats inuntreated areas

Second Post-Treatment MonitoringDate: September 7, 2011The second post-treatment monitoring was conducted to determine if any mortality occurred over the summer months, and more generally to provide additional infor-mation on the status of mussel populations in treated versus untreated areas of the pond. Two SCUBA divers spent one day completing the survey. A total of 42 quad-rats were surveyed in treated (22) and untreated (20) areas of the pond (Figure 2) (note: plot locations from the first post-treatment survey were not used because NHESP did not request a second post-treatment survey until after the first post-treatment survey was completed and the plots had already been removed).

Quadrats were 4.0 m2 (2.0m x 2.0m) frames made of interconnected piping. Mussels were collected from the substrate surface to a depth of 10 cm below the surface. All mussels found within each quadrat were identified to species. For all rare mussel species found, shell length and shell condition were recorded, as well as the water depth and predominant substrate type within the quadrat they were found in.

Quadrat without eastern pondmusselQuadrat with eastern pondmussel

N

4


RESULTS

First Post-Treatment MonitoringCounts: During the post-treatment observation, a com-bined total of 75 mussels (100 percent) were counted in the treatment plots and 73 mussels (97.3 percent) were counted in control plots (Table 2).

Mortality: Eleven of the original 150 test mussels were recorded as dead during the post-treatment survey (93 percent survival). Seven of the dead mussels were in treat-ment plots and four were in control plots. This was not a significant between the treatment plots and control plots.

Embeddedness: Mean embeddedness for all mussels in the three treatment plots was 32.5 percent (means for in-dividual treatment plots were 16.7, 40.2, and 40.5), and in the three control plots it was 34.5 percent (means for individual control plots were 40.2, 22.6, and 40.8). The mean embeddedness for all six plots combined was 33.5 percent. Though there was some variation in embedded-ness, means were very similar between treatment and con-trol plots.

Filtering: The mean filtering score for the treatment plots was 0.37 (means for individual treatment plots were 0.21, 0.37, and 0.52), compared to 0.68 for the control plots (means for individual control plots were 0.48, 0.67, and 0.88). The mean filtering score for all six plots combined was 0.52. Mussels were filtering significantly less in the treatment plots than in the control plots.

Responsiveness: The mean responsiveness score for the treatment plots was 0.64 (means for individual treatment plots were 0.42, 0.72, and 0.79), compared to 0.75 for the control plots (means for individual control plots were 0.57, 0.76. and 0.92). The mean responsiveness score for all six plots combined was 0.69. The difference between treatment and control plots was statistically insignificant.

Second Post-Treatment MonitoringA total of 329 live and 255 dead mussels (shells) were counted during the second post-treatment survey (Ap-pendix 1, Table 3). The abundance and species composi-tion of shells was generally similar to what was observed for live animals. The surveyors found the same four spe-cies that had been seen during earlier surveys.

A total of 196 live mussels were found in the quadrats in untreated areas of the lake, representing a mean den-sity of 2.45 mussels/m2 (range among quadrats = 0.5-12.0 mussels/m2, standard deviation = 2.58). Eastern elliptio comprised 75 percent of all mussels in the untreated areas, followed by eastern lampmussel (15.3 percent), eastern floater (8.7 percent), and eastern pondmussel (1.0 per-cent).

A total of 133 live mussels were found in the quad-rats in the treated areas of the lake, representing a mean density of 1.51 mussels/m2 (range among quadrats = 0.0-5.0 mussels/m2, standard deviation = 1.36). Eastern el-liptio comprised 52.6 percent of all mussels in the treated areas, followed by eastern floater (15.3 percent), eastern lampmussel (14.8 percent), and eastern pondmussel (2.0 percent).

Six live eastern pondmussels and one shell were found during the second post-treatment monitoring (Table 4, see also Figure 2 for locations). Four of the live mussels were found in treated areas of the lake, and two live mus-sels and one shell were found untreated areas of the lake. Shell lengths ranged from 55-85 mm (mean = 70 mm) in the treated areas of the lake. Shell lengths ranged from 75-95 mm (mean = 83 mm) in the untreated areas of the lake.

There was notable spatial variation in the relative abundance of some species; we consider this natural varia-tion that is related to habitat variables rather than chemi-cal treatments or stressors. For example, eastern floater were nearly twice as abundant in the treated areas versus untreated areas, and this was expected because shallower coves with mucky bottoms often support higher densi-

Plot Live Count Dead Count Embeddedness Filtering Score Responsiveness ScoreTreatment 1 24 1 16.7 0.21 0.42Treatment 2 23 2 40.2 0.37 0.72Treatment 3 21 4 40.5 0.52 0.79Control 1 22 3 40.2 0.48 0.57Control 2 22 1 22.6 0.67 0.76Control 3 25 0 40.8 0.88 0.92All Treatment 68 7 32.5 0.37 0.64All Control 69 4 34.5 0.68 0.75All Combined 137 11 33.5 0.52 0.69

Table 2. Summary of mussel counts and scores for embeddedness, filtering, and responsiveness of mussels in treatment plots, control plots, and all plots combined.

5


ties of this species. In contrast, eastern elliptio was nearly twice as abundant in untreated areas, and this was also expected because this species is often more abundant in sand-gravel substrates.

DISCUSSION

We did not detect a significant response of freshwater mussels to the Phycomycin treatment in Oldham Pond. There was some mortality (seven percent) evident during the first post-treatment survey but there was no difference between treatment and control plots. The only measured parameter that suggested a possible negative response to the chemical treatment was a significantly lower filter-ing score in the treatment plots than in the control plots during the first post-treatment survey. Surveyors noted a large amount very fine particulate organic matter and black (presumably decomposing) material settled on the bottom. It is plausible that these were dead plankton and that dissolved oxygen was depressed near the lake bottom, which might have caused mussels to stop or slow filtering, but not die or change behavior in other ways.

The second post-treatment survey documented differ-ences in mussel densities in treated versus untreated areas, but differences were probably due to natural habitat varia-tion rather than a response to the chemical treatment. Overall, there was very little evidence that Phycomycin treatments had any short-term adverse effects on freshwa-

SpeciesElCo LaRa PyCa LiNa All

Parameter Area of the Lake Live Shell Live Shell Live Shell Live Shell Live ShellMussel Counts Untreated (n=20) 147 101 30 7 17 14 2 1 196 123

Treated (n=22) 70 88 29 18 30 26 4 0 133 132Total (n=42) 217 189 59 25 47 40 6 1 329 255

Mussel Density Untreated (n=20) 1.84 1.26 0.38 0.09 0.21 0.18 0.03 0.01 2.45 1.54(mussels/m2) Treated (n=22) 0.80 1.00 0.33 0.20 0.34 0.30 0.05 0.00 1.51 1.50

Total (n=42) 1.29 1.13 0.35 0.15 0.28 0.24 0.04 0.01 1.96 1.52

Table 3. Summary of mussel counts and densities in quadrats in untreated and treated areas of the lake, surveyed during the second post-treatment monitoring. See Appendix 1 for all data.

LiNa Count Shell Length (mm)

ShellCondition

WaterDepth (ft)Latitude Longitude Quadrat Live Shell Substrate

42.0660450 -70.8372190 Untreated 6 1 0 95.0 0.5 Sand, Gravel 3.042.0668220 -70.8362540 Untreated 14 1 0 80.0 0.25 Silt, Sand 5.042.0672210 -70.8360870 Untreated 16 0 1 75.0 1 Silt 4.042.0704750 -70.8406290 Treated 3 1 0 65.0 0.25 Silt, Gravel 5.042.0676480 -70.8429620 Treated 13 1 0 85.0 0.5 Silt, Gravel 5.042.0680760 -70.8415120 Treated 14 1 0 55.0 0.25 Sand, Gravel 4.042.0666170 -70.8419130 Treated 17 1 0 75.0 0.5 Silt, Gravel 5.0

Table 4. Summary of locations, counts, shell length, shell condition, and habitat for eastern pondmussels found during the second post-treatment monitoring.

ter mussels at the doses used in Oldham Pond and under the conditions that prevailed during the treatment pro-cess. The long-term effects of Phycomycin treatment to freshwater mussels were not addressed in this study and remain unknown.

LITERATURE CITED

Biodrawversity. 2009. Freshwater Mussel Survey and Habitat Assessment in Oldham Pond (Hanson and Pembroke, Massachusetts) for a Proposed Chemical Treatment to Control Algae. Report submitted to the Pembroke Watershed Association and Aquatic Control Technology, Inc.

Mattson, M.D., P.J. Godfrey, R.A. Barletta, and A. Aiello. 2004. Eutrophication and Aquatic Plant Management in Massachusetts: Final Generic Environmental Impact Re-port. Edited by Kenneth J. Wagner. Department of En-vironmental Protection and Department of Conserva-tion and Recreation. Executive Office of Environmental Affairs, Commonwealth of Massachusetts.

6


Species

Area of theLake

ElCo LaRa PyCa LiNa All Live Density(mussels/m2)Latitude Longitude Quad# Live Shell Live Shell Live Shell Live Shell Live Shell

42.0653920 -70.8378670 Untreated 1 8 11 4 3 1 0 0 0 13 14 3.2542.0653970 -70.8375810 Untreated 2 2 10 2 0 0 1 0 0 4 11 1.0042.0657060 -70.8377420 Untreated 3 5 2 3 0 0 0 0 0 8 2 2.0042.0656410 -70.8375490 Untreated 4 0 0 2 0 1 0 0 0 3 0 0.7542.0660780 -70.8375640 Untreated 5 1 9 0 0 1 1 0 0 2 10 0.5042.0660450 -70.8372190 Untreated 6 8 3 4 0 2 2 1 0 15 5 3.7542.0664650 -70.8374170 Untreated 7 4 0 2 0 1 0 0 0 7 0 1.7542.0664120 -70.8371040 Untreated 8 1 11 2 0 1 1 0 0 4 12 1.0042.0666330 -70.8371000 Untreated 9 9 3 2 0 0 0 0 0 11 3 2.7542.0667770 -70.8373260 Untreated 10 6 21 0 0 0 0 0 0 6 21 1.5042.0669830 -70.8372030 Untreated 11 3 0 0 0 0 0 0 0 3 0 0.7542.0669180 -70.8370370 Untreated 12 3 2 0 0 0 1 0 0 3 3 0.7542.0668560 -70.8365240 Untreated 13 5 0 2 0 0 0 0 0 7 0 1.7542.0668220 -70.8362540 Untreated 14 2 0 0 0 0 0 1 0 3 0 0.7542.0672270 -70.8365270 Untreated 15 2 10 2 0 1 3 0 0 5 13 1.2542.0672210 -70.8360870 Untreated 16 4 2 0 0 2 1 0 1 6 4 1.5042.0624310 -70.8309170 Untreated 17 15 4 0 0 2 1 0 0 17 5 4.2542.0628170 -70.8321330 Untreated 18 11 4 1 1 0 0 0 0 12 5 3.0042.0618730 -70.8312690 Untreated 19 41 5 3 2 4 2 0 0 48 9 12.0042.0624740 -70.8320820 Untreated 20 17 4 1 1 1 1 0 0 19 6 4.7542.0703920 -70.8402980 Treated 1 1 2 0 0 0 0 0 0 1 2 0.2542.0704630 -70.8403910 Treated 2 3 1 3 1 1 2 0 0 7 4 1.7542.0704750 -70.8406290 Treated 3 1 3 2 1 2 1 1 0 6 5 1.5042.0703780 -70.8407150 Treated 4 1 8 0 0 0 0 0 0 1 8 0.2542.0706530 -70.8409370 Treated 5 2 1 3 1 3 1 0 0 8 3 2.0042.0705780 -70.8410400 Treated 6 1 7 1 2 3 2 0 0 5 11 1.2542.0690250 -70.8440510 Treated 7 0 0 0 0 0 0 0 0 0 0 0.0042.0692970 -70.8433570 Treated 8 0 1 0 0 0 0 0 0 0 1 0.0042.0683900 -70.8437650 Treated 9 0 0 0 0 1 1 0 0 1 1 0.2542.0688820 -70.8429560 Treated 10 0 0 0 0 0 0 0 0 0 0 0.0042.0679780 -70.8433870 Treated 11 4 1 2 2 2 3 0 0 8 6 2.0042.0684900 -70.8423110 Treated 12 0 6 0 0 0 0 0 0 0 6 0.0042.0676480 -70.8429620 Treated 13 3 2 0 0 4 2 1 0 8 4 2.0042.0680760 -70.8415120 Treated 14 5 2 3 2 2 3 1 0 11 7 2.7542.0671320 -70.8425500 Treated 15 7 6 1 1 0 1 0 0 8 8 2.0042.0677540 -70.8412490 Treated 16 6 7 4 1 3 4 0 0 13 12 3.2542.0666170 -70.8419130 Treated 17 10 7 4 2 5 3 1 0 20 12 5.0042.0673900 -70.8410160 Treated 18 13 29 1 1 0 0 0 0 14 30 3.5042.0657440 -70.8409980 Treated 19 0 0 0 0 0 0 0 0 0 0 0.0042.0663650 -70.8403190 Treated 20 2 1 1 2 2 1 0 0 5 4 1.2542.0655880 -70.8407790 Treated 21 5 2 3 1 2 2 0 0 10 5 2.5042.0659440 -70.8402180 Treated 22 6 2 1 1 0 0 0 0 7 3 1.75

APPENDIX 1Locations and mussel counts (live and shells) in the 42 quadrats surveyed

during the second post-treatment monitoring.

APPENDIX D

Pembroke Watershed Association Volunteer Temperature & Dissolved Oxygen Data

2011 OLDHAM POND VOLUNTEER TEMPERATURE AND DISSOLVED OXYGEN DATA

Sample Site Collection Date Secchi Depth (in.) Depth (ft.) Temperature (C) Dissolved Oxygen (mg/l)1 6/19/2011 24 4 23 9.95

7 22.9 9.3311 20.6 5.28

2 6/19/2011 30 4 22.8 10.157 20.8 5.67

11 20.1 3.363 6/19/2011 30 4 22.3 9.56

7 20.4 3.2

Sample Site Collection Date Secchi Depth (in.) Depth (ft.) Temperature (C) Dissolved Oxygen (mg/l)1 7/6/2011 37 4 25.5 9.25

7 24.1 6.72 7/6/2011 39 4 25.1 7.65

7 24.1 6.813 7/6/2011 45 4 26.4 10.48

7 23.7 5.65


8 25.3 2.2511 22.7 0.77

2 7/20/2011 31 4 26.7 6.868 25.5 3.08

11 23.3 0.323 7/20/2011 32 4 26.8 7.74

7 25.7 3.68

PRE-DAWN COLLECTION (5:30 AM)Sample Site Collection Date Secchi Depth (in.) Depth (ft.) Temperature (C) Dissolved Oxygen (mg/l)

1 7/21/2011 31 4 26 7.667 25.9 5.6

11 24 1.392 7/21/2011 30 4 26.5 7.38

7 26.5 6.4910 25 1.7

3 7/21/2011 30 4 26.8 7.277 25.9 3.05


7 26.4 6.1711 25.6 1.1

2 8/3/2011 35 4 26.3 5.747 26.2 4.05

11 25.3 0.53 8/3/2011 30 4 26 5.21

7 25.7 1.94

Page 1

2011 OLDHAM POND VOLUNTEER TEMPERATURE AND DISSOLVED OXYGEN DATA

PRE-DAWN COLLECTION (6:00 AM)Sample Site Collection Date Secchi Depth (in.) Depth (ft.) Temperature (C) Dissolved Oxygen (mg/l)

1 8/19/2011 38 4 24 8.727 23.4 5.99

11 23 3.452 8/19/2011 36 4 24.4 7.55

7 24.2 6.6111 23.1 2.37

3 8/19/2011 31 4 24.6 6.848 23.6 2.73


7 24 5.72 8/24/2011 24 4 24.2 6.16

7 24.3 6.13 8/24/2011 30 4 24 6.05

7 24 5.1

Page 2

APPENDIX E

Algae Toxin Sampling Results

Greenwater Labs

Anatoxin-a/Microcystin/Saxitoxin Data Report Project: Aquatic Control Tech – Oldham Pond

Sample Identification Sample Collection Date

Oldham Pond (beach) 110706

Toxins – Anatoxin-a (ANTX-A), microcystin (MC), saxitoxin (STX)

Sample Prep – The sample was ultra-sonicated to lyse all cells and release toxins. Solid phase

extraction (SPE) was also utilized for anatoxin-a extraction and preconcentration (100x).

Duplicate samples were spiked (Lab Fortified Matrix, LFM) with 1.0 µg/L MCLR, 0.1 µg/L

ANTX-A and 0.2 µg/L STX which provided quantitative capability and additional qualitative

confirmation.

Analytical Methodology – Liquid chromatography/ mass spectrometry/ mass spectrometry

(LC/MS/MS) was utilized for the determination of ANTX-A. The [M+H]+ ion for ANTX-A

(m/z 166) was fragmented and the major product ions (m/z 149, 131, 107, and 91) provided both

specificity and sensitivity. The current methodology established a detection limit of 0.05 µg/L

and a quantification limit of 0.1 µg/L for ANTX-A.

A microcystins enzyme linked immunosorbent assay (ELISA) was utilized for the quantitative

and sensitive congener-independent detection of MCs. The current assay is sensitive to down to

a LOD/LOQ of 0.15 µg/L for total MCs. The average recovery of a laboratory fortified blank

(LFB) spiked with 1 µg/L MCLR was 91%.

A saxitoxin enzyme linked immunosorbent assay (ELISA) was utilized for the quantitative

detection of saxitoxin. The current assay is sensitive down to a LOD/LOQ of 0.05 µg/L

saxitoxin. The average recovery of an LFB spiked with 0.2 µg/L STX was 100%.

Summary of ANTX-A/MC/STX Results

Sample ANTX-A levels MC level STX level

( µg/L) (µg/L) (µg/L)

Oldham Pond (beach) ND ND ND

ND = Not detected above the detection limit

Detection Limit = 0.05 µg/L (ANTX-A), 0.15 µg/L (MC), 0.05 µg/L (STX)

Limit of Quantification = 0.1 µg/L (ANTX-A), 0.15 µg/L (MC), 0.05 µg/L (STX)

Submitted by: _________________

Mark T. Aubel, Ph.D.

Date: 7/8/11

GreenWater Laboratories Contact: [email protected]

205 Zeagler Drive [email protected]

Suite 302

Palatka FL 32177

Ph (386) 328-0882

Fax (386) 328-0882

Tested on: 7/7/2011

Method: Enzyme-Linked ImmunoSorbent Assay (ELISA)

Analyte: Microcystins

Analyzed by: Amanda Foss

Sample ID/ Initial Conc. Dilution Assay Final Dilution Avg. LFB Avg. LFM Final Average

Date Collected Factor Ratio Value, ug/L Factor Recovery(%) Recovery (%) Concentration (ug/L) (ug/L)

Oldham Pond Beach 1x none ND 1 91 96 ND ND

7/6/11 1x none ND 1 91 96 ND

Aquatic Control Tech., Inc.

MICROCYSTIN RESULTS

7/6/11 1x none ND 1 91 96 ND

ND = Not detected above LOD/LOQ

LOD/LOQ = 0.15 µg/L

LFB = 1.0 µg/L MCLR

LFM = 1.0 µg/L MCLR

Submitted to: Aquatic Control Tech

Submitted by: ____________________ Gerald N Smith

Mark T. Aubel, Ph.D. 11 John Rd

Date: 7/7/2011 Sutton MA 01590

Ph (508) 865-1000

GreenWater Laboratories Contact: [email protected]

205 Zeagler Drive [email protected]

Suite 302

Palatka FL 32177

Ph (386) 328-0882

Fax (386) 328-0882

Tested on: 7/8/2011

Method: Enzyme-Linked ImmunoSorbent Assay (ELISA)

Analyte: Saxitoxin

Analyzed by: Amanda Foss

Sample ID/ Initial Conc. Dilution Assay Final Dilution Avg. LFB Avg. LFM Final Average

Date Collected Factor Ratio Value, ug/L Factor Recovery(%) Recovery (%) Concentration (ug/L) (ug/L)

Aquatic Control Tech., Inc.

SAXITOXIN RESULTS

Oldham Pond Beach 1x none ND 1 100 90 ND ND

7/6/11 1x none ND 1 100 90 ND

ND = Not detected above LOD/LOQ

LOD/LOQ = 0.05 µg/L

LFB = 0.2 µg/L CYN

LFM = 0.2 µg/L CYN

Submitted by: ____________________ Submitted to: Aquatic Control Tech

Mark T. Aubel, Ph.D. Gerald N Smith

Date: 7/8/2011 11 John Rd

Sutton MA 01590

Ph (508) 865-1000

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