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Observing systems, modeling, and forecasting

Predicting harmful algal blooms (HABs) requires integrating physical, chemical, and biological data collected from observing networks and then assimilating these data into models, which are used to generate forecasts. In 2005, the Harmful Algal Research and Response: A National Environmental Science Strategy 2005-2015 (HARRNESS, 2005) made recommendations on how to improve HAB modeling and forecasting over the next decade. Key HARRNESS recommendations related to sensing, networking, and modeling HABs included: ● Support the development and validation of new and improved technologies for remote cell and toxin detection, and for modeling and forecasting, ● Improve coordination of monitoring/ and modeling efforts, both at national and regional levels, ● Improve the use of networking technologies for monitoring and modeling efforts, ● Conduct sustained time series measurements of the biotic, chemical, and physical environments impacted by HABs, ● Develop food web models on the ecosystem fate and effects of toxins, ● Develop and improve species-specific models that link to physical-biological models. Here we review HAB observing, modeling, and forecasting advances and technologies and recommend research and management priorities for the next decade and beyond. Our report encompasses sensing technologies, sensor networking and data management, models and forecasts, and the paths to operationalize forecasts. Continued improvements of deployable sensors are foundational to improving early warning indicators, models, and forecasts, which are only as good as the underlying data. Sensing technology has advanced considerably in the last decade; for example, more capable fluorometric pigment sensors can track changes in bloom biomass in real-time. Additionally, automated imaging/classifying systems to identify and quantify key harmful algal (HA) taxa are being routinely deployed. However, deployable toxin sensors are available for only some HAB toxins and continue to be identified as a critical need by researchers and managers. As more and improved sensors and technologies become available, the data quality associated with each sensor needs to be assessed. Data quality encompasses the reliability, accuracy, and uncertainty associated with sensor-generated data. These data need to be of known quality so that researchers, managers, and end-users can reliably determine if the information is appropriate for their intended applications. Many of the data quality recommendations from HARRNESS (2005) are still relevant and have been reiterated within the management community. Understanding and documenting data quality, and when applicable, standardizing best practices for sensor use, continue to be recommended.

Report

Could blackbird mortality from avicide DRC-1339 contribute to avian botulism outbreaks in North Dakota?

Blackbird (family lcteridae) depredation on sunflower (Helianthus annuus) crops in the prairie states of the United States has motivated the proposed use of an avicide, DRC-1339 (3-chloro-4-methylaniline), to decrease their numbers. The resulting mortality of blackbirds at wetland roosts could increase the potential of avian botulism occurring in affected marshes. To assess this possibility, we seeded (artificially placed) blackbird carcasses in selected wetlands in Stutsman County, North Dakota, during August-September 2000 and July-September 2001 to evaluate their rate of decomposition and role in initiating avian botulism outbreaks. We monitored carcasses to determine their persistence, the frequency and amount of maggots produced, and the presence of type C botulinum toxin. In 10 of our 12 study wetlands, blackbird carcasses were not rapidly removed by scavengers, thus providing substrate for maggot growth and potential production of Clostridium botulinum toxin. Decomposition of carcasses occurred rapidly, and maggot production averaged 4a??5 g per carcass within 9 days. We were unable to detect C. botulinum type C toxin in any of the 377 blackbird carcasses or the 112 samples of maggots we collected in 2000 or 2001. None of the 25 blackbird carcasses we tested contained botulinum spores, the most probable explanation for the absence of botulinum toxin production. Our results indicate that the likelihood of DRC-1339-poisoned blackbirds causing botulism outbreaks would be minimal in North Dakota wetlands during late summer and early autumn.

North Dakota, South Dakota

Detection of microcystin and other cyanotoxins in lakes at Isle Royale National Park, Pictured Rocks National Lakeshore, and Sleeping Bear Dunes National Lakeshore, northern Michigan, 2012–13

Although cyanotoxins released during algal blooms have become an increasing concern in surface waters across the United States, the presence of cyanotoxins in northern Michigan lakes had not been evaluated in detail. The U.S. Geological Survey and National Park Service (NPS) led a 2-year study (2012 and 2013) to determine the presence of microcystin and other algal toxins in several inland lakes at Isle Royale National Park (hereafter referred to as ISRO, Pictured Rocks National Lakeshore (hereafter referred to as PIRO), and Sleeping Bear Dunes National Lakeshore (hereafter referred to as SLBE). Samples also were collected at four sites in Lake Michigan within the SLBE. The two analytical techniques used in the study were enzyme-linked immunosorbent assays (ELISA) for microcystin, cylindrospermopsin, and saxitoxin; and liquid chromatography/tandem mass spectrometry (LC/MS/MS) for a larger suite of algal toxins. Neither cylindrospermopsin nor saxitoxin were detected in the 211 samples. Microcystin was detected in 31 percent of samples (65 of 211 samples) analyzed by the ELISA method, but no sample results exceeded the World Health Organization recreational health advisory standard for microcystin (10 micrograms per liter [µg/L]). However, about 10 percent of the samples (21 of 211 samples) that were collected from PIRO and SLBE and were analyzed by ELISA for microcystin had concentrations greater than the U.S. Environmental Protection Agency (EPA) drinking water 10-day health advisory of 0.3 µg/L for children preschool age and younger (less than 6-years old). One sample collected in 2012 from SLBE exceeded the EPA drinking water 10-day health advisory of 1.6 µg/L for school-age children through adults (6-years old and older). In 2012, the highest concentration of 2.7 µg/L was detected in Florence Lake within SLBE. Many visitors enjoy recreation in or on the water and camp in the backcountry at these national parks where the most common source of drinking water is filtered surface water. Approximately 18 percent of the samples (39 of 211 samples) were analyzed by LC/MS/MS to confirm the ELISA results and to evaluate the samples for a larger suite of algal toxins. In general, the microcystin results between the ELISA and LC/MS/MS methods were similar; although, the ELISA results tended to be slightly higher than the summation of LC/MS/MS microcystin congeners. The slightly higher ELISA results might be because the ELISA microcystin method is reactive with the ADDA functional group common to all microcystins, and because not all microcystin congeners are included in the LC/MS/MS method. The LC/MS/MS method indicated that the congener microcystin-LR was the most frequently detected, followed by microcystin-WR and microcystin-YR. Sixteen of the lakes included in this study also were monitored by the NPS for nutrients. Total phosphorus (TP) concentrations were, on average, highest at the ISRO lakes, whereas total nitrogen (TN) concentrations were highest at SLBE. The average annual TN:TP ratios for the 16 lakes within the national park and national lakeshores ranged from ratios of 20 to 89. Overall, results indicated a slight increase in percentage of microcystin detections with an increase in the TN:TP ratio (R-squared 0.269 and 0.340, respectively [2012 and 2013 combined dataset] derived from linear regression). This study also indicated that even in the absence of visible algal blooms, microcystin may be present. Most microcystin concentrations did not exceed the EPA’s 10-day health advisory drinking-water benchmark. In general, these results provide a useful baseline with which to evaluate potential future changes in algal toxin concentrations.

Michigan

Environmental characterization of Blue Mesa Reservoir and potential causes of and management strategies for harmful algal blooms, 1970 through 2023, Curecanti National Recreation Area, Colorado

Blue Mesa Reservoir, in the Curecanti National Recreation Area, is the largest storage reservoir in Colorado and consists of three distinct basins: Iola (the shallowest), Cebolla, and Sapinero. After algal toxins were first documented in Iola basin in 2018, the U.S. Geological Survey began a study in cooperation with the National Park Service, Colorado River Water Conservation District, Upper Gunnison River Water Conservancy District, Gunnison County, Project 7 Water Authority, and Uncompahgre Valley Water Users Association to better understand occurrence of toxic cyanobacteria harmful algal blooms (cyanoHABs) and identify possible causal mechanisms to potentially inform management strategies. Toxic cyanoHABS occurred when the algal toxin microcystin exceeded a concentration of 8 micrograms per liter primarily in Iola basin in 2018 and 2020–22, years having some of the lowest reservoir water-level elevations (reservoir levels) since 1984. The toxic cyanoHABs started in mid-September and continued through the fall months. Algal abundance was greatest in Iola basin compared to Cebolla and Sapinero basins, with Aphanizomenon , a toxin-producing cyanobacterium, being the most abundant. During blooms, enhanced algal photosynthesis caused elevated pH and dissolved oxygen concentrations especially in Iola basin. Continuous monitor data in Iola basin indicated peaks in phycocyanin fluorescence, pH, and dissolved oxygen concentration that preceded the onset of toxic cyanoHABs by about 2 weeks potentially indicating a useful early warning monitoring strategy for future response to toxic cyanoHABs. Long-term trends showed increases in mean air and surface-water temperatures and chlorophyll- a concentrations in the reservoir but no change in nutrient inputs from major tributaries. In Iola basin, reservoir level was positively correlated with Secchi disk depth and inversely correlated with total phosphorus concentration. Because of its shallow depth, the effect of low reservoir levels may disproportionately affect Iola basin compared to other basins, resulting in algal blooms and toxin production especially at reservoir levels below about 7,470 feet above North American Vertical Datum of 1988. Elevated phosphorus at low reservoir level likely was primarily phosphorus contained in algal tissue. This report indicates that the main driver for recent toxic cyanoHABs in Iola basin is low reservoir level that likely causes favorable conditions (shallow and warm) for algal growth and increased recruitment of algae from bottom sediments such as during wind-driven turbulence. Control of external nutrients to the reservoir is unlikely to help control algal blooms because Aphanizomenon fixes nitrogen from the atmosphere, and there is an abundant geogenic source of phosphorus. Maintenance of reservoir levels greater than about 7,470 feet might help minimize the occurrence of toxic cyanoHABs. Additional data could help better understand how the timing and duration of reservoir levels below 7,470 feet contribute to toxic cyanoHABs.

Colorado

Immunological evidence of variation in exposure and immune response to Bacillus anthracis in herbivores of Kruger and Etosha National Parks

Exposure and immunity to generalist pathogens differ among host species and vary across spatial scales. Anthrax, caused by a multi-host bacterial pathogen, Bacillus anthracis , is enzootic in Kruger National Park (KNP), South Africa and Etosha National Park (ENP), Namibia. These parks share many of the same potential host species, yet the main anthrax host in one (greater kudu ( Tragelaphus strepsiceros ) in KNP and plains zebra ( Equus quagga ) in ENP) is only a minor host in the other. We investigated species and spatial patterns in anthrax mortalities, B. anthracis exposure, and the ability to neutralize the anthrax lethal toxin to determine if observed host mortality differences between locations could be attributed to population-level variation in pathogen exposure and/or immune response. Using serum collected from zebra and kudu in high and low incidence areas of each park (18- 20 samples/species/area), we estimated pathogen exposure from anti-protective antigen (PA) antibody response using enzyme-linked immunosorbent assay (ELISA) and lethal toxin neutralization with a toxin neutralization assay (TNA). Serological evidence of pathogen exposure followed mortality patterns within each system (kudus: 95% positive in KNP versus 40% in ENP; zebras: 83% positive in ENP versus 63% in KNP). Animals in the high-incidence area of KNP had higher anti-PA responses than those in the low-incidence area, but there were no significant differences in exposure by area within ENP. Toxin neutralizing ability was higher for host populations with lower exposure prevalence, i.e., higher in ENP kudus and KNP zebras than their conspecifics in the other park. These results indicate that host species differ in their exposure to and adaptive immunity against B. anthracis in the two parks. These patterns may be due to environmental differences such as vegetation, rainfall patterns, landscape or forage availability between these systems and their interplay with host behavior (foraging or other risky behaviors), resulting in differences in exposure frequency and dose, and hence immune response.

Etosha National Park, Kruger National Park

Waterbird mortality from botulism type E in Lake Michigan: An update

Three outbreaks of botulism type E occurring in waterbirds on Lake Michigan since autumn 1976 are discussed. Natural ingestion of food containing type E toxin by Ring-billed Gulls ( Larus delawarensis ) and the presence of type E toxin in blood from moribund gulls were demonstrated. Concurrent presence of type C and type E botulinal toxins was found in a die-off of Common Loons ( Gavia immer ). In combination with previous reported outbreaks, these incidents suggest that this disease is geographically widespread in Lake Michigan, and that environmental conditions conducive to type E botulinal toxin production and consumption occur in both summer and autumn.

Lake Michigan

PCR und ELISA - Alternativen zum Maustest für die Analyse des Botulismus-Neurotoxin-C1 Giftbildungspotentiales in Umweltproben? [PCR and ELISA - in vitro alternatives to the mouse-bioassay for assessing the botulinum-neurotoxin-C1 production potential in environmental samples?]

Botulism is one of the most important bird diseases world-wide and is caused by the intoxication with Botulinum-Neurotoxin-C1 (BoNt-C1), which is produced by toxigenic clostridia under appropriate conditions. Avian botulism leads regularly to large losses among the migrating bird populations breeding and resting at the saltwater pools of the Austrian national park Neusiedler See-Seewinkel. Despite of its ethical dubiousness and its high technical expense the mouse-bioassay is still used as the routine standard method for the detection of BoNt-C1. According to the 3R-concept, in vitro alternative methods for the qualitative detection of BoNt-C1 (immunostick-ELISA) and a corresponding BoNt-C1 gene fragment (nested-PCR) were established. In order to estimate the BoNt-C1 production potential the methods were tested with sediment samples from different saltwater pools subjected to cultivation conditions appropriate for in vitro BoNt-C1-production. With the mouse-bioassay, 52 out of 77 samples were found to have a positive toxin production potential. The immunostick-ELISA showed a similar sensitivity as the mouse-bioassay and exhibited a highly significant positive correlation (r=0.94; p<0.001) with the mouse-bioassay in detecting BoNt-C1. The nested-PCR approach revealed higher numbers of positive BoNt-C1 gene fragment detections as compared to the direct toxin analysis approaches. A weak correlation (r=0.21; p=0.07) with the mouse-bioassay was discernible, no correlation was found with the immunostick-ELISA (r=0.09; p=0.46). Obviously, the PCR approach detected the BoNt-C1 gene fragment in some of the samples where no toxin expression has occurred. Thus it is suggested that the qualitative immunostick-ELISA represents a potential in vitro alternative to the mouse-bioassay for assessing the BoNt-C1 production potential in environmental samples. In contrast, qualitative BoNt-C1 gene fragment detection via PCR led to an overestimation of the actual toxin production potential.

Neusiedler See-Seewinkel National Park

Demonstration, validation, and application of hyperspectral microscopy for the collection of cyanobacterial spectral signatures

Cyanobacterial and other algal blooms are an environmental concern in waterbodies worldwide. While these blooms are a nuisance for recreational activities, they can also be harmful to human and wildlife health when the algae produce and release toxins. Algal community composition can be monitored and analyzed by acquiring hyperspectral images that provide information on various photosynthetic and accessory pigments. Validated, traceable measurements are needed to compare data collected by different hyperspectral instruments. In this proof-of-concept study, we detail the development and validation of a custom hyperspectral microscopy imaging system and assess whether this technology can differentiate between cyanobacteria genera based on differences in their reflectance characteristics. As not all cyanobacteria produce toxins, the ability to distinguish among taxa could be used to identify potential toxin-producers and guide field sampling and further research. Spectral characterization of these taxa contributes to remote sensing efforts to characterize and identify cyanobacterial genera at larger spatial scales.

Oregon

A chronicle of a killer alga in the west: Ecology, assessment, and management of Prymnesium parvum blooms

Since the mid-1980s, fish-killing blooms of Prymnesium parvum spread throughout the USA. In the south central USA, P. parvum blooms have commonly spanned hundreds of kilometers. There is much evidence that physiological stress brought on by inorganic nutrient limitation enhances toxicity. Other factors influence toxin production as well, such as stress experienced at low salinity and temperature. A better understanding of toxin production by P. parvum remains elusive and the identities and functions of chemicals produced are unclear. This limits our understanding of factors that facilitated the spread of P. parvum blooms. In the south central USA, not only is there evidence that the spread of blooms was controlled, in part, by migration limitation. But there are also observations that suggest changed environmental conditions, primarily salinity, facilitated the spread of blooms. Other factors that might have played a role include altered hydrology and nutrient loading. Changes in water hardness, herbicide use, system pH, and the presence of toxin-resistant and/or P. parvum -inhibiting plankton may also have played a role. Management of P. parvum in natural systems has yet to be attempted, but may be guided by successes achieved in small impoundments and mesocosm experiments that employed various chemical and hydraulic control approaches.

Hydrobiologia

Estimating microcystin levels at recreational sites in western Lake Erie and Ohio

Cyanobacterial harmful algal blooms (cyanoHABs) and associated toxins, such as microcystin, are a major global water-quality issue. Water-resource managers need tools to quickly predict when and where toxin-producing cyanoHABs will occur. This could be done by using site-specific models that estimate the potential for elevated toxin concentrations that cause public health concerns. With this study, samples were collected at three Ohio lakes to identify environmental and water-quality factors to develop linear-regression models to estimate microcystin levels. Measures of the algal community (phycocyanin, cyanobacterial biovolume, and cyanobacterial gene concentrations) and pH were most strongly correlated with microcystin concentrations. Cyanobacterial genes were quantified for general cyanobacteria, general Microcystis and Dolichospermum , and for microcystin synthetase ( mcyE ) for Microcystis , Dolichospermum , and Planktothrix. For phycocyanin, the relations were different between sites and were different between hand-held measurements on-site and nearby continuous monitor measurements for the same site. Continuous measurements of parameters such as phycocyanin, pH, and temperature over multiple days showed the highest correlations to microcystin concentrations. The development of models with high R 2 values (0.81–0.90), sensitivities (92%), and specificities (100%) for estimating microcystin concentrations above or below the Ohio Recreational Public Health Advisory level of 6 μg L −1 was demonstrated for one site; these statistics may change as more data are collected in subsequent years. This study showed that models could be developed for estimates of exceeding a microcystin threshold concentration at a recreational freshwater lake site, with potential to expand their use to provide relevant public health information to water resource managers and the public for both recreational and drinking waters.

Ohio

Identifying the origin of waterbird carcasses in Lake Michigan using a neural network source tracking model

Avian botulism type E is responsible for extensive waterbird mortality on the Great Lakes, yet the actual site of toxin exposure remains unclear. Beached carcasses are often used to describe the spatial aspects of botulism mortality outbreaks, but lack specificity of offshore toxin source locations. We detail methodology for developing a neural network model used for predicting waterbird carcass motions in response to wind, wave, and current forcing, in lieu of a complex analytical relationship. This empirically trained model uses current velocity, wind velocity, significant wave height, and wave peak period in Lake Michigan simulated by the Great Lakes Coastal Forecasting System. A detailed procedure is further developed to use the model for back-tracing waterbird carcasses found on beaches in various parts of Lake Michigan, which was validated using drift data for radiomarked common loon ( Gavia immer ) carcasses deployed at a variety of locations in northern Lake Michigan during September and October of 2013. The back-tracing model was further used on 22 non-radiomarked common loon carcasses found along the shoreline of northern Lake Michigan in October and November of 2012. The model-estimated origins of those cases pointed to some common source locations offshore that coincide with concentrations of common loons observed during aerial surveys. The neural network source tracking model provides a promising approach for identifying locations of botulinum neurotoxin type E intoxication and, in turn, contributes to developing an understanding of the dynamics of toxin production and possible trophic transfer pathways.

Lake Michigan

Large differences in herbivore performance emerge from simple herbivore behaviors and fine-scale spatial heterogeneity in phytochemistry

Patterns of phytochemistry localisation in plant tissues are diverse within and across leaves. These spatial heterogeneities are important to the fitness of herbivores, but their effects on herbivore foraging and dietary experience remain elusive. We manipulated the spatial variance and clusteredness of a plant toxin in a synthetic diet landscape on which individual caterpillars fed. We monitored caterpillars with cameras across most of their larval development. Caterpillars that fed on diets with a lower spatial variance and more clustered arrangement of toxins had overall worse performance, mostly because those caterpillars ate less, moved more, ingested more toxin, or failed to physiologically acclimate. Using empirically parameterised individual-based models, we found that differences in movement away from, not towards, less toxic food drove a body size-dependent effect of clusteredness. Hence, the spatial pattern of phytochemicals itself, beyond mean concentration, can have important consequences for herbivores through complex interactions with herbivore foraging.

Ecology Letters

Toxicoinfectious botulism in commercial caponized chickens

During the summer of 2003, two flocks of commercial broiler chickens experienced unusually high death losses following caponizing at 3 wk of age and again between 8 and 14 wk of age. In September, fifteen 11-wk-old live capons were submitted to the Iowa State University Veterinary Diagnostic Laboratory for assistance. In both flocks, the second episode of elevated mortality was associated with incoordination, flaccid paralysis of leg, wing, and neck muscles, a recumbent body posture characterized by neck extension, and diarrhea. No macroscopic or microscopic lesions were detected in affected chickens. Hearts containing clotted blood and ceca were submitted to the National Wildlife Health Center in Madison, WI. Type C botulinum toxin was identified in heart blood and ceca by mouse bioassay tests. Enzyme-linked immunosorbent assay tests on heart blood samples were also positive for type C botulinum toxin. Clostridium botulinum was isolated from the ceca and genes encoding type C botulinum toxin were detected in cecal contents by a polymerase chain reaction test. Chickens are less susceptible to botulism as they age, and this disease has not previously been documented in broilers as old as 14 wk of age. Wound contamination by spores of C. botulinum may have contributed to the unusually high death losses following caponizing.

Avian Diseases

Relations between DNA- and RNA-based molecular methods for cyanobacteria and microcystin concentration at Maumee Bay State Park Lakeside Beach, Oregon, Ohio, 2012

Water samples were collected from Maumee Bay State Park Lakeside Beach, Oregon, Ohio, during the 2012 recreational season and analyzed for selected cyanobacteria gene sequences by DNA-based quantitative polymerase chain reaction (qPCR) and RNA-based quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Results from the four DNA assays (for quantifying total cyanobacteria, total Microcystis , and Microcystis and Planktothrix strains that possess the microcystin synthetase E ( mcyE ) gene) and two RNA assays (for quantifying Microcystis and Planktothrix genera that are expressing the microcystin synthetase E ( mcyE ) gene) were compared to microcystin concentration results determined by an enzyme-linked immunosorbent assay (ELISA). Concentrations of the target in replicate analyses were log10 transformed. The average value of differences in log10 concentrations for the replicates that had at least one detection were found to range from 0.05 to >0.37 copy per 100 milliliters (copy/100 mL) for DNA-based methods and from >0.04 to >0.17 copy/100 mL for RNA-based methods. RNA has a shorter half-life than DNA; consequently, a 24-hour holding-time study was done to determine the effects of holding time on RNA concentrations. Holding-time comparisons for the RNA-based Microcystis toxin mcyE assay showed reductions in the number of copies per 100 milliliters over 24 hours. The log difference between time 2 hours and time 24 hours was >0.37 copy/100 mL, which was higher than the analytical variability (log difference of >0.17 copy/100 mL). Spearman’s correlation analysis indicated that microcystin toxin concentrations were moderately to highly related to DNA-based assay results for total cyanobacteria (rho=0.69), total Microcystis (rho=0.74), and Microcystis strains that possess the mcyE gene (rho=0.81). Microcystin toxin concentrations were strongly related with RNA-based assay results for Microcystis mcyE gene expression (rho=0.95). Correlation analysis could not be done for Planktothrix mcyE gene expression because of too few detections.

Ohio

Toxic algae in inland waters of the conterminous United States—A review and synthesis

Cyanobacteria are the most common toxigenic algae in inland waters. Their toxins can affect the health of aquatic and terrestrial organisms, including humans. Other algal groups, such as haptophytes (e.g., Prymnesium parvum ) and euglenoids (e.g., Euglena sanguinea ), can also form harmful algal blooms (HABs) whose toxins cause injury to aquatic biota but currently have no known effects on human health. Prymnesium parvum , however, is responsible for some of the worst HAB-related ecological disasters recorded in inland waters. Here, we provide an overview of the primary toxigenic algae found in U.S. inland waters: cyanobacteria (planktonic forms), P. parvum , and E. sanguinea with the objective of describing their similarities and differences in the areas of HAB ecology, algal toxins, and the potential for future range expansion of HABs. A detailed account of bloom habitats and their known associations with land cover and use is provided from the perspective of water quality. This review revealed that salinity may have an influence on inland cyanobacterial blooms and cyanotoxins that had not been fully recognized previously.

Water

Preliminary evaluation of a simple in vitro test for the diagnosis of type C botulism in wild birds

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of type C botulinum toxin ( Clostridium botulinum ) in wild birds. This simple, antigen-capture ELISA utilizes polystyrene immunosticks as the solid substrate, chicken antitoxin (IgY) as the coating antibody, rabbit antitoxin as the primary antibody, and peroxidase-labeled goat-anti-rabbit as the secondary antibody. To evaluate the immunostick ELISA as a diagnostic test for avian botulism, known concentrations of toxin were added to heparinized blood collected from healthy birds and tested by both the ELISA and mouse bioassay. Also, blood samples from 236 bird carcasses submitted to the National Wildlife Health Center (NWHC) for cause of death determinations were tested by both procedures. Using ≤ 0.5 ml as the test volume for both procedures, the ELISA was less sensitive, detecting 0.25 ng/ml of toxin compared to 0.12 ng/ml for the mouse bioassay. Using the same volume of test sample for diagnostic submissions (≤ 0.5 ml), the ELISA was positive for 60% of the 149 clinically-diagnosed cases of botulism, whereas the mouse bioassay was positive for 79%. However, we demonstrated that with larger sample volumes (≥ 1.0 ml), the sensitivity of the ELISA may be equivalent or better than the mouse test due to the concentrating effect of the ELISA procedure. These preliminary results suggest that when adequate sample volumes are available, the immunostick ELISA can replace the mouse test for the diagnosis of botulism in wild birds.

Journal of Wildlife Diseases

Efficacy of a type C botulism vaccine in green-winged teal

We tested the efficacy of a single dose of Botumink toxoid for protecting wild green-winged teal (Anas crecca) during botulism epizootics caused by Clostridium botulinum type C. We challenged control and immunized ducks with four different doses of type C botulinum toxin to determine the LD50 for this species and to evaluate vaccine protection. Fewer immunized ducks were affected with botulism than control ducks, indicating that a single dose of Botumink toxoid could increase the survival of ducks during epizootics. However, the frequency of immunized ducks with signs of botulism increased with the challenge dose of botulinum toxin. Even at doses of botulinum toxin approximately 2 to 4 green-winged teal LD50, about 50% of the immunized ducks were affected. We believe an improved vaccine or a better delivery system is required to justify immunization of wild birds for experimental survival studies.

California, Utah

Avian botulism: geographic expansion of a historic disease

Avian botulism is a paralytic, often fatal disease of birds resulting from ingestion of toxin produced by the bacterium Clostridium botulinum. Waterfowl die-offs from the botulism are usually caused by type C toxin; sporadic die-offs among fish-eating birds, such as common loons (Gavia immer) and gulls, have been caused by type E toxin.

Fish and Wildlife Leaflet