INVESTIGATION OF NITRATE VALUES AT THE HARPSWELL SOUND BUOY: A COMPARISON OF METHODS
Anderson, Amy, Bowdoin College, firstname.lastname@example.org
Roesler, Collin, Bowdoin College, 6800 College Station, Brunswick ME 04011, email@example.com
Nutrients provide the basic building blocks for any species to live. Phytoplankton, the single-celled aquatic photosynthesizer, requires phosphate and nitrogen compounds to meet their basic needs. Thus, the dynamics of phytoplankton growth in the natural environment can be controlled by nutrient availability. Nutrient dynamics are complicated, but by examining the major sources of nitrogen (in the form of nitrate and nitrite), we are exploring the role nutrients play in the occurrences and blooms of Alexandrium fundyense, the harmful algal species that causes paralytic shellfish poisoning (PSP) in coastal Maine and is commonly known as red tide. We focused our study in Harpswell Sound, a sentinel site for A. fundyense appearance and early closures for PSP toxicity in shellfish.
Research completed over the summer aimed to accurately estimate the concentration of nitrate remotely using the absorption spectrum. In the laboratory, we developed standard curves relating nitrate concentration to absorption peak height by chemically reconstructing seawater to understand its complexities. This will allow us to determine of natural nutrient concentration and to shed light on nutrient dynamics.
In understanding nutrient dynamics, we can apply this knowledge to occurrences of A. fundyense. As a shellfish ingests the organism, it stores the toxin within its tissues and the concentration of the toxin expands with increased exposure. The A. fundyense toxin progresses to other species with consumption of poisoned shellfish and can result in fatalities. By gaining information about this detrimental organism’s environment, we have greater insight into its development.
A SPATIAL ANALYSIS OF ALS IN NORTHERN NEW ENGLAND
Caller, Tracie A. Department of Neurology, Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA, caller@Hitchcock.org
Carlos, Heather A. Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA, Heather.A. Carlos@ Dartmouth.edu
Doolin, James Department of Neurology, Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA, firstname.lastname@example.org
Chipman, Jonathan W. Department of Geography, Dartmouth College, Hanover, NH, USA, Jonathan.W.Chipman@ Dartmouth.edu
Shi, Xun Department of Geography, Dartmouth College, Hanover, NH, USA, Xun.Shi@Dartmouth.edu
Harris, Brent T. Department of Neurology, Georgetown University, Washington, DC, USA
Stommel, Elijah W. Department of Neurology, Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA, Stommel@Hitchcock.org
Background: An environmental factor which triggers sporadic ALS is supported by geographic disparities in the incidence of ALS and by the development of the disease in conjugal couples. A high incidence of ALS has been documented in Guam, and the cyanobacteria toxin beta-methylamino-L-alanine (BMAA), found in components of the Chamorro diet, has been implicated. It is unknown whether true disease clusters of ALS exist in Northern New England, and if present, if they might be associated with toxic cyanobacterial blooms that occur in New England water bodies. This study aims to elucidate geographic variation in ALS prevalence within Northern New England, and to potentially evaluate for a spatial association with cyanobacteria blooms.
Methods: We reviewed electronic records and community databases to identify the dwelling addresses of patients presenting with ALS between 1998 and 2009 in New Hampshire (NH), Vermont (VT), and Maine (ME), adjusting for the underlying population density. Using ArcGIS software for spatial analysis, we evaluated for significant clustering and spatial outliers with Anselin's Local Moran’s I test.
Results: We identified clusters of high prevalence in NH, ME, and VT which are statistically significant. All were within close proximity of documented cyanobacterial blooms.
Discussion: There appears to be areas of significant spatial clustering within NH and VT. It is uncertain whether living within close proximity of a cyanobacteria bloom is an independent risk factor for developing ALS. Further analysis will be needed to confirm whether there is any correlation between these areas of clustering and potential environmental risk factors.
CYANOBACTERIAL TOXINS FOUND IN UPPER KLAMATH LAKE, OREGON: IMPLICATIONS FOR ENDANGERED FISH
Kathy Echols1, Tamara Wood2, Chris Ottinger3, Barry Rosen5, Summer Burdick4, Scott Vanderkooi4
1US Geological Survey, BRD, Columbia Environmental Research Center, Columbia MO, USA,2US Geological Survey, WRD, WRD District Office, Portland OR, USA,3US Geological Survey, BRD, Leetown Science Center, National Fish Health Research Laboratory, Kearneysville WV, USA,4US Geological Survey, Klamath Falls Field Station, Klamath Falls, OR, USA, 5US Geological Survey, Florida Integrated Science Center, Orlando, Florida, USA
Upper Klamath Lake is a large hyper-eutrophic lake, found on the east side of the Cascade Mountains in Southern Oregon. Conditions in the lake include high levels of nitrogen and phosphorus nutrients that facilitate large, continuous cycles of cyanobacterial blooms from late spring through the fall. These cyanobacterial blooms include species of algae that are known to produce toxins that adversely affect other aquatic organisms. This study was designed to evaluate the presence of these toxins over three summer seasons (2007 - 2009) and to determine if there is any risk to the two endangered fish in the system: the Lost River sucker (Deltistes luxatus) and the shortnose sucker (Chasmistes brevirostris). Juvenile fish of these species are rare and appear to have poor survival in this ecosystem. Water and algae samples were collected monthly in 2007; bi-weekly in 2008 and weekly in 2009 from late June through September. Collected water/algal samples were filtered, extracted and analyzed using an enzyme-linked immunoassay (ELISA) method for quantitation of microcystin toxins, in water and algae. In 2007, high levels of microcystin toxins (up to 17.4 ug/L) were found in algae late in the summer and evidence from fish pathology showed exposure and adverse effects from these toxins. In 2008, lower levels of total microcystin were found in water/algal samples; from a low of 0.17 ug/L to a high concentration of 6.1 ug/L. The high water concentration for total microcystin in 2009 was 83 ug/L. Published LD50s for fish are around 1,700 ug/kg for one microcystin toxin variant; so assuming that juvenile fish are consuming 1% of their body mass in toxic algae, the average exposure to fish from microcystin was assessed to be approximately 1.5-fold greater than the published effects levels of microcystins to fish in 2007 and 0.7-fold greater in 2008 and 1.4-fold greater in 2009. Thus, cyanobacterial toxins may be limiting survival of juvenile endangered Lost River sucker and shortnose sucker in Upper Klamath Lake.
Moldaenke1, C., Dahlhaus1 A. , Dahlhaus1, H., Hoffmüller1, J.,1bbe Moldaenke, Germany, email@example.com ; PP Systems, Amesbury, MA, USA, firstname.lastname@example.org
Carpentier2, C.2Environmental Institute, The Netherlands, email@example.com
The bbe AlgaeTorch is a new instrument to measure algae in a water column. It displays the total content of chlorophyll as well as the chlorophyll content of cyanobacteria. This instrument enables the surveillance of lakes and rivers and has recently been recommended as the principle instrument to serve as an early warning system for water quality of public swimming areas regarding cyanobacteria.
The bbe BenthoTorch, launched last year, is an instrument to measure benthic algae in water, on river banks and even on walls. It can determine the total chlorophyll content per cm² and discriminate between green algae, diatoms and cyanobacteria. New correction methods for different substrates assist in the correction of non-linear data and reflection properties.
MARINE CYANOBACTERIA FROM THE COLOMBIAN CARIBBEAN: CHARACTERIZATION AND ASSESSMENT OF ECOLOGICAL ROLES
Prato, Julian, Universidad Jorge Tadeo Lozano, 301 Granada St, Fort Pierce FL, firstname.lastname@example.org
Puyana, Mónica, Universidad Jorge Tadeo Lozano, Cra 4 # 22-16 Bogotá, Colombia, email@example.com
Dobretsov, Sergey, Sultan Qaboos University, Muscat, Oman, firstname.lastname@example.org
Valerie Paul, Smithsonian Marine Station, 701 Seaway drive, Fort Pierce FL. Paul@si.edu
Blooms of marine benthic cyanobacteria have become increasingly frequent in many reef areas worldwide. We have started a survey of benthic cyanobacteria blooms in the Colombian Caribbean in order to determine species involved, intensity, magnitude, bloom formation dynamics, secondary metabolite composition and bioactivity as well as assessment of their potential ecological roles. We have surveyed several blooms in the Colombian Caribbean and have identified as the most representative species in bloom formation the species Symploca hydnoides, Phormidium submembranaceum, Lyngbya majuscula, L. sordida, Oscillatoria margaritifera, O. nigroviridis and O. acuminata. The cyanobacterial mats we collected were never composed of just one species, and up to five species coexisted in a mat. We have performed feeding preference assays of cyanobacterial crude extracts on the anaspidean gastropod Bursatella leachii, finding a general avoidance of Lyngbya spp., S. hydnoides and P. submembra naceum extracts. We have also assessed some potential ecological roles against microorganisms. Some cyanobacterial extracts are strong inhibitors of the filamentous marine fungus Dendryphiella salina at concentrations of 1 mg/ml. Some cyanobacterial extracts have also shown quorum sensing inhibition in the bacterium Chromobacterium violaceum strain CV17, in concentrations in the range 0.8-2.1 mg ml-1.
DEVELOPMENT AND VALIDATION SENSITIVE IMMUNOASSAY FORMATS FOR MICROCYSTIN DETECTION
Fernando Rubio (email@example.com), Lisa Kamp, Jennifer Church.
Abraxis LLC, 54 Steamwhistle Drive, Warminster, PA 18974, USA
Cyanobacteria (blue-green-algae) are known to produce various toxins that can be assigned to different substance classes, such as alkaloids, lipopolysaccharides, polyketides, or peptides. Cyanobacteria live in terrestrial, fresh, brackish, or marine water. Some of the toxins they produce can be toxic, others can cause severe taste and odor problems in drinking water supplies. Cyanobacterial toxins can make drinking water and recreational use of water unsafe. Animals die yearly as a result of cyanotoxins, and though human death is not commom, many people experience symptoms indicative of cyanotoxin exposure. Very little is known about the long-term side effects of ingestion of cyanotoxins, although there is a guideline set by WHO for safe concentrations, minimal concentrations could cause an effect over time.
Concerns about contamination of lakes and reservoirs with algal toxins have led to the need for more rapid, sensitive, and selective methods of analysis.
Immunoassays (ELISAs) have proven to be rapid, sensitive, accurate, and cost-effective. Microtiter plate ELISAs have previously been described and widely applied to the detection of pesticides and other environmental contaminants in various sample matrices, including water, soil, produce, and fish tissue. Other immunoassay formats utilizing immunochromatography are often single step devices where a test sample is analyzed for the presence of certain analytes. For example, a specified volume of the sample is contacted with one end of a test strip. The test strip contains colored particles coated with a binder dried on the strip. As the sample is wicked up the test strip, the analyte in the sample reacts with the binder coated on the particles. The test strip also contains antigens in discrete zones. As the reaction mixture flows up the strip, any reaction between the antigens and the analyte, if present, may be observed by the appearance or non-appearance of color in the zon es.
This paper describes the development and assay performance conducted under the U.S. EPA Environmental Technology Testing Verification (ETV) of a microtiter plate ELISAs, and an immuno-chromatography device for the quick analysis of microcystins in drinking, and recreational water samples. The results showed that the ELISAs exhibit excellent sensitivities (0.1 ppb). Data obtained with water samples during the U.S. EPA Verification Program as well as sample comparison will also be presented.
QUÉBEC’S VALIDATION STUDY OF MICROCYSTINS SCREENING STRIPS
Sinotte, Marc, Ministère du Développement Durable, de l’Environnement et des Parcs (MDDEP), Édifice Marie-Guyart, 7e étage, 675, boulevard René-Lévesque Est, Québec (Québec) G1R 5V7, firstname.lastname@example.org
Within Québec’s ‘’Management Plan for Cyanobacterial Bloom Episodes’’(MPCBE), about 1000-1400 samples are analysed annually for Microcystin (MC) by HPLC-MS/MS. These analyses are costly and are putting some burden on the lab technicians due to a mandatory turnover of 48 hours. MDDEP regional offices technicians are sampling water bodies following individual phone calls reporting the blooms, the majority of which are free of toxins. In order to reduce cost and concentrate on samples at greater risk MDDEP started a validation study of two different kits of Microcystin strip test against HPLC-MS/MS. The study, with a double blind protocol, was done under the actual MPCBE and analysed about 500 different samples so that statistical precision could be achieved. Both kits proved to be helpful to screen out samples not at risk, without sacrificing public health protection.
WET LABS EFFORTS IN IN SITU TOXIN MEASUREMENT
Bruno Soffientino and Andrew Barnard, WET Labs Inc. 70 Dean Knauss Road, Narragansett, RI 02882-1197
Toxic algal blooms are challenging to study and monitor because they display large spatial and temporal variability. It is generally accepted that in situ monitoring by autonomous instrument packages has the potential to overcome the costs and logistic difficulties associated with observing these events. While some organismal features, such as pigment fluorescence, can be measured in situ, no autonomous instrument commercially available today has the capability to measure harmful algal toxins.
With funding under the National Ocean Partnership Program (NOPP), WET Labs has undertaken proof-of-principle work towards the development of a compact, simple, and relatively low-cost autonomous instrument to measure dissolved toxins in situ. We demonstrated that a paramagnetic bead-based immunoassay is a workable approach towards an in-situ instrument to measure toxins, and other organic chemicals of interest. Immunoassays (ELISA ) are available for a variety of toxins, and some are being approved for health-related environmental monitoring; magnetic beads have been used in several immunoassay automation approaches (flow cytometric, microfluidic). In collaboration with a NOAA biotoxin laboratory (Dr. Vera Trainer, Seattle, WA) we converted an established immunoassay for domoic acid to the bead-based format, and we tested it with fluorescence detection. At the same time we tested different methods of storing and dispensing the beads automatically. Current work focuses on assembling a breadboard prototype that can run the proof-of-principle assay.
ARE CURRENT GUIDELINES FOR DRINKING WATER REALLY SAFE FOR DEVELOPING ORGANISM? AN EXPLORATORY STUDY
Larissa Takser, Université de Sherbrooke, email@example.com
Alexander Suvorov, Université de Sherbrooke
The number of blue algae blooms in Canadian water sources increases every year. Cyanotoxins pollution is public health concern because of their high toxicity potential and low degradability. There is increasing evidence on neurotoxic potential of major cyanotoxins. Our objective was to explore behavioural effects of cyanotoxins at guideline concentrations in rodent developmental model. Pregnant and lactating dams were exposed to microcystin-MR, cylindrospermopsin, anatoxin A through dringking water. Their offspring were tested for spontaneous locomotor activity, anxiety, glucose tolerance, liver function parameters. Our data show the increase in locomotor activity in male offsprings and the increase in liver enzyme ALT in all pups. Our findings suggests that the safety of current drinking water guidelines for cyanotoxins should be considered as matter of concern for developing offspring.
OPERATIONALIZING WATER QUALITY INFORMATION FROM NASA SATELITE PLATFORMS WITH WEB-GIS TECHNOLOGIES, MOBILE TOOLS AND CLOUD COMPUTING
Nathan Torbick, PhD
Applied Geosolutions, LLC
Newmarket, NH 03857
Pressures on water resources will continue to increase in the future. More than half the world’s population lives adjacent to water bodies and carry out activities that cause pressures and increase aquatic stressors such as anthropogenic eutrophication and algal blooms. Increasing concern over cyanobacteria and human health has arisen as threats to and from water resources are recognized. However, current approaches to assess water quality rely heavily on expensive point sampling with limited spatiotemporal coverage. In addition, scales of existing monitoring strategies do not meet the extensive temporal and spatial scales needed to address water quality – public health relationships. The goal of this project is to operationalize water quality indicators using NASA satellites platforms and cloud computing for all inland water bodies in CONUS by 2014. We extract the strengths of multiple NASA satellite sensors to generate a scaled product set ranging from lake temperature to phycocyanin-rich cyanobacteria indices. Information can be obtained using mobile technologies and web-GIS frameworks.
AN IN SITU NUTRIENT-ADDITION PHYTOPLANKTON BIOASSAY STUDY OF WEST MONPONSETT POND
Zoto, George A. Ph.D. Mattson, Mark D. PhD.; Bureau of Resource Protection, MassDEP, 20 River 20 Riverside Drive, Lakeville, MA 02347, firstname.lastname@example.org
An in situ bioassay study was initiated at West Monponsett Pond, Halifax, MA using the natural assemblage of the phytoplankton community of this pond to assess the effect of nitrogen, phosphorus, and other nutrients on the dynamics of this algal community. As a shallow, eutrophic, 303(d) listed impaired water body, West Monponsett Pond is also a class A water body that receives nutrient inputs from from numerous sources. Ultimately, as part of a TMDL related nutrient management strategy for use in controlling bluegreen algal blooms, the outcome of this research will provide guidance to the Department in its decision making regarding nutrient management strategies for restoring water quality as part of TMDL planning and implementation. This study reports the response of the following major cyanobacteria taxa: Cuspidothrix issatschenkoi (previously known as Aphanizomenon issatschenkoi), Aphanizomenon, Raphidiopsis cf. mediterranea, Oscillatoria, Planktolyngbya limnetica, Microcystis, and Coelosphaerium), as well as green algae and diatoms, to each of the nutrient additions (N, P, NP, NP + trace) and presents its quantitative findings as cell density (cells/mL), biomass (cubic microns/mL), and chlorophyll a.
In support of recently published studies, this study highlights the difficulty encountered distinguishing between Cuspidothrix, and Raphidiopsis. The plasticity of the morphology of these bluegreens (changes in cell shape within the filament and at its apical location as well as the presence/absence of heterocysts/akinetes) have been described in the literature and also identified in this study, suggesting that these changes may be the genetic response to nutrients and other environmental factors. Note worthy are recent studies that propose that Raphidiopsis mediterranea may represent a non-heterocytous life-cycle stages of Cylindropermopsis raciborskii – an invasive toxic bluegreen alga whose range is extending northward and yet to be found in Massachusetts.