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Total protein concentration and diagnostic test results for gray wolf ( Canis lupus ) serum using Nobuto filter paper strips

Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves ( Canis lupus ) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

Journal of Wildlife Diseases↗

Nonlethal screening of bat-wing skin with the use of ultraviolet fluorescence to detect lesions indicative of white-nose syndrome

Definitive diagnosis of the bat disease white-nose syndrome (WNS) requires histologic analysis to identify the cutaneous erosions caused by the fungal pathogen Pseudogymnoascus [formerly Geomyces] destructans (Pd). Gross visual inspection does not distinguish bats with or without WNS, and no nonlethal, on-site, preliminary screening methods are available for WNS in bats. We demonstrate that long-wave ultraviolet (UV) light (wavelength 368–385 nm) elicits a distinct orange–yellow fluorescence in bat-wing membranes (skin) that corresponds directly with the fungal cupping erosions in histologic sections of skin that are the current gold standard for diagnosis of WNS. Between March 2009 and April 2012, wing membranes from 168 North American bat carcasses submitted to the U.S. Geological Survey National Wildlife Health Center were examined with the use of both UV light and histology. Comparison of these techniques showed that 98.8% of the bats with foci of orange–yellow wing fluorescence (n = 80) were WNS-positive based on histologic diagnosis; bat wings that did not fluoresce under UV light (n = 88) were all histologically negative for WNS lesions. Punch biopsy samples as small as 3 mm taken from areas of wing with UV fluorescence were effective for identifying lesions diagnostic for WNS by histopathology. In a nonlethal biopsy-based study of 62 bats sampled (4-mm diameter) in hibernacula of the Czech Republic during 2012, 95.5% of fluorescent (n = 22) and 100% of nonfluorescent (n = 40) wing samples were confirmed by histopathology to be WNS positive and negative, respectively. This evidence supports use of long-wave UV light as a nonlethal and field-applicable method to screen bats for lesions indicative of WNS. Further, UV fluorescence can be used to guide targeted, nonlethal biopsy sampling for follow-up molecular testing, fungal culture analysis, and histologic confirmation of WNS.

Journal of Wildlife Diseases↗

Spatial and temporal patterns of avian paramyxovirus-1 outbreaks in Double-Crested Cormorants ( Phalacrocorax auritus ) in the USA

Morbidity and mortality events caused by avian paramyxovirus-1 (APMV-1) in Double-crested Cormorant (DCCO; Phalacrocorax auritus ) nesting colonies in the US and Canada have been sporadically documented in the literature. We describe APMV-1 associated outbreaks in DCCO in the US from the first reported occurrence in 1992 through 2012. The frequency of APMV-1 outbreaks has increased in the US over the last decade, but the majority of events have continued to occur in DCCO colonies in the Midwestern states. Although morbidity and mortality in conesting species has been frequently reported during DCCO APMV-1 outbreaks, our results suggest that isolation of APMV-1 is uncommon in species other than DCCO during APMV-1 outbreaks and that the cause of mortality in other species is associated with other pathogens. Populations of DCCO do not appear to have been significantly affected by this disease; however, because at least 65% of the APMV-1 outbreaks in DCCO in the US have involved APMV-1 strains classified as virulent to poultry (virulent Newcastle disease virus), its persistence and increased occurrence in DCCO warrants continued research and surveillance.

Journal of Wildlife Diseases↗

Retrospective analysis of the epidemiologic literature, 1990–2015, on wildlife-associated diseases from the Republic of Korea

To assess the status of research on wildlife diseases in the Republic of Korea (ROK) and to identify trends, knowledge gaps, and directions for future research, we reviewed epidemiologic publications on wildlife-associated diseases in the ROK. We identified a relatively small but rapidly increasing body of literature. The majority of publications were focused on public or livestock health and relatively few addressed wildlife health. Most studies that focused on human and livestock health were cross-sectional whereas wildlife health studies were mostly case reports. Fifteen diseases notifiable to the World Organisation for Animal Health were identified and 21 diseases were identified as notifiable to either the Korean Ministry of Health, Welfare, and Family Affairs or the Korean Ministry of Agriculture. Two diseases were reported as occurring as epidemics; highly pathogenic avian influenza (HPAI) and virulent Newcastle disease. Six diseases or disease agents were described in the literature as emerging including HPAI, rabies, Babesia microti , avian coronaviruses, scrub typhus, and severe fever thrombocytopenia syndrome virus. The diseases for which there were the largest number of publications were HPAI and rabies. The majority of wildlife-associated zoonotic disease publications focused on food-borne parasitic infections or rodent-associated diseases. Several publications focused on the potential of wildlife as reservoirs of livestock diseases; in particular, water deer ( Hydropotes inermis ) and wild boar ( Sus scrofa ). In contrast, there were few publications on diseases of concern for wildlife populations or research to understand the impacts of these diseases for wildlife management. Increased focus on prospective studies would enhance understanding of disease dynamics in wildlife populations. For the high-consequence diseases that impact multiple sectors, a One Health approach, with coordination among the public health, agricultural, and environmental sectors, would be important. This type of review can provide useful information for countries or regions planning or implementing national wildlife health programs.

Journal of Wildlife Diseases↗

Prevalence and distribution of Wellfleet Bay virus exposure in the Common Eider ( Somateria mollissima )

Between 1998 and 2014, recurrent mortality events were reported in the Dresser's subspecies of the Common Eider ( Somateria mollissima dresseri ) on Cape Cod, Massachusetts, USA near Wellfleet Harbor. The early die-offs were attributed to parasitism and emaciation, but beginning in 2006 a suite of distinct lesions was observed concomitant with the isolation of a previously unknown RNA virus. This novel pathogen was identified as an orthomyxovirus in the genus Quaranjavirus and was named Wellfleet Bay virus (WFBV). To assess evidence of exposure to this virus in Common Eiders, we conducted a longitudinal study of the prevalence of WFBV antibodies at multiple locations from 2004–14; we collected 2,258 serum samples from six locations and analyzed each using a microneutralization assay. Results corroborate the emergence of WFBV in 2006 based on the first detection of antibodies in that year. Significantly higher prevalence was detected in Common Eiders sampled in Massachusetts compared to those in Maine, Nova Scotia, and Québec. For birds breeding and wintering in Massachusetss, viral exposure varied by age, sex, and season of sampling, and prevalence by season and sex were highly interrelated with greater numbers of antibody-positive males in the autumn and females in the spring. No evidence of viral exposure was detected in the Northern subspecies ( Somateria mollissima borealis ). Among the locations sampled, Massachusetts appears to be the epicenter of Common Eider exposure to WFBV. Further research is warranted to understand the factors controlling the epidemiology of WFBV in Massachussetts, including those that may be limiting geographic expansion of this virus.

Journal of Wildlife Diseases↗

A serosurvey of diseases of free-ranging gray wolves (Canis lupus) in Minnesota

We tested serum samples from 387 free-ranging wolves ( Canis lupus ) from 2007 to 2013 for exposure to eight canid pathogens to establish baseline data on disease prevalence and spatial distribution in Minnesota's wolf population. We found high exposure to canine adenoviruses 1 and 2 (88% adults, 45% pups), canine parvovirus (82% adults, 24% pups), and Lyme disease (76% adults, 39% pups). Sixty-six percent of adults and 36% of pups exhibited exposure to the protozoan parasite Neospora caninum . Exposure to arboviruses was confirmed, including West Nile virus (37% adults, 18% pups) and eastern equine encephalitis (3% adults). Exposure rates were lower for canine distemper (19% adults, 5% pups) and heartworm (7% adults, 3% pups). Significant spatial trends were observed in wolves exposed to canine parvovirus and Lyme disease. Serologic data do not confirm clinical disease, but better understanding of disease ecology of wolves can provide valuable insight into wildlife population dynamics and improve management of these species.

Minnesota↗

Mortality trends in northern sea otters (Enhydra lutris kenyoni) collected from the coasts of Washington and Oregon (2002–15)

During 2002−15 we examined the causes of mortality in a population of northern sea otters ( Enhydra lutris kenyoni ). Beachcast sea otters were collected primarily from the coast of Washington. Although there are no permanent sea otter residents in Oregon, several beachcast otters were collected from the Oregon coast. Infectious diseases were the primary cause of death (56%) for otters we examined. Sarcocystosis was the leading infectious cause of death (54%) and was observed throughout the study period. Some infectious diseases, such as morbilliviral encephalitis and leptospirosis, were documented for a limited number of years and then not detected again despite continued testing for these pathogens in necropsied animals. Trauma was the second most common cause of death (14%) during the study period. The continued stable growth of the Washington population of otters suggests they are able to tolerate current mortality rates.

Oregon, Washington↗

Exposure of Alaska brown bears (Ursus arctos) to bacterial, viral, and parasitic agents varies spatiotemporally and may be influenced by age

We collected blood and serum from 155 brown bears ( Ursus arctos ) inhabiting five locations in Alaska during 2013–16 and tested samples for evidence of prior exposure to a suite of bacterial, viral, and parasitic agents. Antibody seroprevalence among Alaska brown bears was estimated to be 15% for Brucella spp., 10% for Francisella tularensis , 7% for Leptospira spp., 18% for canine adenovirus type 1 (CAV-1), 5% for canine distemper virus (CDV), 5% for canine parvovirus, 5% for influenza A virus (IAV), and 44% for Toxoplasma gondii . No samples were seropositive for antibodies to Trichinella spp. Point estimates of prior exposure to pathogens among brown bears at previously unsampled locations generally fell within the range of estimates for previously or contemporaneously sampled bears in Alaska. Statistical support was found for variation in antibody seroprevalence among bears by location or age cohort for CAV-1, CDV, IAV, and Toxoplasma gondii . There was limited concordance in comparisons between our results and previous serosurveys regarding spatial and age-related trends in antibody seroprevalence among Alaska brown bears suggestive of temporal variation. However, we found evidence that the seroprevalence of CAV-1 antibodies is consistently high in bears inhabiting SW Alaska and the cumulative probability of exposure may increase with age. We found evidence for seroconversion or seroreversion to six different infectious agents in one or more bears. Results of this study increase our collective understanding of disease risk to both Alaska brown bear populations and humans that utilize this resource.

Alaska↗

Long-term survival of Pseudogymnoascus destructans at elevated temperatures

White-nose syndrome is an emerging fungal disease that has devastated hibernating bat populations across eastern North America. The causal pathogen, Pseudogymnoascus destructans (PD), is a psychrophilic fungus with a known maximal growth temperature of 20 C. Although it is widely speculated that PD is primarily spread between hibernacula by the movement of bats, experimental evidence is lacking to demonstrate that PD can endure temperatures experienced by active bats for periods of time that would facilitate dispersal of viable fungus. We used an in vitro culture-based approach to study the survival of PD conidia on three artificial growth media and bat fur. The fungus was incubated at three temperatures it might realistically be exposed to on nonhibernating bats or in the environment outside of caves and mines (24 C, 30 C, and 37 C). When incubated on artificial media, we found that PD conidia were able to survive for a maximum of 150 d when exposed to temperatures of 24 C, 60 d at 30 C, and 15 d at 37 C. At all temperatures, maximal survival duration was recorded when conidia were incubated on brain–heart infusion agar with 10% volume of sheep ( Ovis aries ) blood. When incubated on bat fur, viable PD was recovered at 180 d, 60 d, and 5 d when exposed to temperatures of 24 C, 30 C, and 37 C, respectively. Our results suggest that viable PD conidia may be able to survive on or within the bodies of bats, which may facilitate long-distance dispersal. The long-term viability of the fungus on various fomites may differ, and therefore must be assessed for each potential substrate.

Journal of Wildlife Diseases↗

Poecivirus is present in individuals with beak deformities in seven species of North American birds

Avian keratin disorder (AKD), a disease of unknown etiology characterized by debilitating beak overgrowth, has increasingly affected wild bird populations since the 1990s. A novel picornavirus, poecivirus, is closely correlated with disease status in Black-capped Chickadees ( Poecile atricapillus ) in Alaska. However, our knowledge of the relationship between poecivirus and beak deformities in other species and other geographic areas remains limited. The growing geographic scope and number of species affected by AKD-like beak deformities require a better understanding of the causative agent to evaluate the population-level impacts of this epizootic. Here, we tested eight individuals from six avian species with AKD-consistent deformities for the presence of poecivirus: Mew Gull ( Larus canus ), Hairy Woodpecker ( Picoides villosus ), Black-billed Magpie ( Pica hudsonia ), American Crow ( Corvus brachyrhynchos ), Red-breasted Nuthatch ( Sitta canadensis ), and Blackpoll Warbler ( Setophaga striata ). The birds were sampled in Alaska and Maine (1999−2016). We used targeted PCR followed by Sanger sequencing to test for the presence of poecivirus in each specimen and to obtain viral genome sequence from virus-positive host individuals. We detected poecivirus in all individuals tested, but not in negative controls (water and tissue samples). Furthermore, we used unbiased metagenomic sequencing to test for the presence of other pathogens in six of these specimens (Hairy Woodpecker, two American Crows, two Red-breasted Nuthatches, Blackpoll Warbler). This analysis yielded additional viral sequences from several specimens, including the complete coding region of poecivirus from one Red-breasted Nuthatch, which we confirmed via targeted PCR followed by Sanger sequencing. This study demonstrates that poecivirus is present in individuals with AKD-consistent deformities from six avian species other than Black-capped Chickadee. While further investigation will be required to explore whether there exists a causal link between this virus and AKD, this study demonstrates that poecivirus is not geographically restricted to Alaska, but rather occurs elsewhere in North America.

Alaska, Maine↗

Pathology of Lagovirus europaeus GI.2/RHDV2/b (rabbit hemorrhagic disease virus 2) in native North American lagomorphs

Rabbit hemorrhagic disease, a notifiable foreign animal disease in the US, was reported for the first time in wild native North American lagomorphs in April 2020 in the southwestern US. Affected species included the desert cottontail ( Sylvilagus audubonii ), mountain cottontail ( Sylvilagus nuttallii ), black-tailed jackrabbit ( Lepus californicus ), and antelope jackrabbit ( Lepus alleni ). Desert cottontails ( n =7) and black-tailed jackrabbits ( n =7) collected in April and May 2020 were necropsied at the US Geological Survey National Wildlife Health Center and tested positive for Lagovirus europaeus GI.2, also known as rabbit hemorrhagic disease virus 2 (GI.2/RHDV2/b), by real-time PCR at the US Department of Agriculture's Foreign Animal Disease Diagnostic Laboratory. Gross and microscopic lesions were similar to those reported in European rabbits ( Oryctolagus cuniculus ) and other hare ( Lepus ) species with GI.2/RHDV2/b infection; they included epistaxis (12/13; 92%); massive hepatocellular dissociation (14/14; 100%) and necrosis or apoptosis (11/11; 100%); pulmonary congestion (12/12; 100%), edema (12/13; 92%), and hemorrhage (11/12; 92%); and acute renal tubular injury (3/8; 38%). As in previous reports, massive hepatocellular dissociation and necrosis or apoptosis were the most diagnostically distinct finding. As North American Sylvilagus and Lepus species appear to be susceptible to fatal GI.2/RHDV2/b infection, additional work is needed to understand the host range, pathogenicity, and potential population effects of GI.2/RHDV2/b in North America.

Arizona, New Mexico, Texas↗

Epistylis spp. infestation in two species of mud turtles (Kinosternon spp.) in the American Southwest

The protistan genus Epistylis contains freshwater colonial species that attach to aquatic organisms in an epibiotic or parasitic relationship. They are known to attach to the epidermis and shells of aquatic turtles, but have not been reported to cause heavy infestations or morbidity in turtles. We documented heavy infestations of Epistylis spp. in several populations of Sonoran mud turtles ( Kinosternon sonoriense ) inhabiting livestock ponds in Arizona, USA, and rough-footed mud turtles ( Kinosternon hirtipes ) from livestock ponds in Texas, USA, over the course of several years. Severe Epistylis spp. infestations on mud turtles appeared to alter diving and swimming behavior when compared to uninfested conspecifics. Infestations were cleared in captivity using tap water or a 10% salt solution, and the turtles had no permanent damage to their shell or epidermis upon clearing. While several of the mud turtles we observed had poor body condition, it is possible that the severe infestations we observed were caused by a comorbidity associated with a pathogen, parasite, or poor habitat quality that made the turtles more susceptible to the Epistylis spp. infestation. Further research on causes for these severe infestations are warranted because they contribute to changes in behavior of the heavily infested turtles and may contribute to morbidity in Kinosternon spp. when mud turtles inhabit extremely warm, shallow, eutrophic aquatic habitats, such as livestock ponds.

Arizona↗

Toxoplasma gondii: Challenges and perspectives in interpreting longitudinal seroprevalence data for a chronic parasitic infection

Toxoplasma gondii —the causative agent of toxoplasmosis—is a zoonotic pathogen of warm-blooded hosts. Infection causes mild-to-severe symptoms, including lethargy, fever, muscle pain, abortion, ocular disease, and encephalitis. Toxoplasma affects many vertebrate species, although felids are the only known definitive hosts. Seroprevalence in wildlife is often assessed using cross-sectional data, but few studies have tracked individual-level infections through time. We present a 4-yr dataset from white-tailed deer ( Odocoileus virginianus ) with repeated sampling of individuals that highlights challenges associated with assigning serostatus to individuals. Using a modified agglutination test, we observed seroconversion from seronegative to seropositive within individuals, as expected. Although toxoplasmosis is known to be a chronic disease, we also found reversion from seropositive to seronegative. Accurate assignment of serostatus is necessary for evaluating effects of infection on behavioral and physiologic outcomes. However, longitudinal data from individuals whose titers oscillate around the positive threshold present novel challenges. Therefore, we discuss the implications for assigning serostatus for chronic toxoplasmosis infection for three proposed approaches: 1) ever positive, always positive; 2) negative until positive and then always positive; and 3) status by sampling period. Clarifying which approach is used to assign serostatus when analyzing longitudinal T. gondii data may enable more meaningful comparisons across systems and studies.

Journal of Wildlife Diseases↗

Investigating microbial size classes associated with the transmission of stony coral tissue loss disease (SCTLD)

Effective treatment and prevention of any disease necessitates knowledge of the causative agent, yet the causative agents of most coral diseases remain unknown, in part due to the difficulty of distinguishing the pathogenic microbe(s) among the complex microbial backdrop of coral hosts. Stony coral tissue loss disease (SCTLD) is a particularly destructive disease of unknown etiology, capable of transmitting through the water column and killing entire colonies within a matter of weeks. Here we used a previously described method to (i) isolate diseased and apparently healthy coral colonies within individual mesocosms containing filtered seawater with low microbial background levels; (ii) incubate for several days to enrich the water with coral-shed microbes; (iii) use tangential-flow filtration to concentrate the microbial community in the mesocosm water; and then (iv) filter the resulting concentrate through a sequential series of different pore-sized filters. To investigate the size class of microorganism(s) associated with SCTLD transmission, we used 0.8 µm pore size filters to capture microeukaryotes and expelled zooxanthellae, 0.22 µm pore size filters to capture bacteria and large viruses, and 0.025 µm pore size filters to capture smaller viruses. In an attempt to further refine which size fraction(s) contained the transmissible element of SCTLD, we then applied these filters to healthy “receiver” coral fragments and monitored them for the onset of SCTLD signs over three separate experimental runs. However, several factors outside of our control confounded the transmission results, rendering them inconclusive. As the bulk of prior studies of SCTLD in coral tissues have primarily investigated the associated bacterial community, we chose to characterize the prokaryotic community associated with all mesocosm 0.22 µm pore size filters using Illumina sequencing of the V4 region of the 16S rRNA gene. We identified overlaps with prior SCTLD studies, including the presence of numerous previously identified SCTLD bioindicators within our mesocosms. The identification in our mesocosms of specific bacterial amplicon sequence variants that also appear across prior studies spanning different collection years, geographic regions, source material, and coral species, suggests that bacteria may play some role in the disease.

PeerJ↗

Status and trends of North American bats: Summer occupancy analysis 2010-2019

• We developed an analytical pipeline supported by web-based infrastructure for integrating continental scale bat monitoring data (stationary acoustic, mobile acoustic, and capture records) to estimate summer (May 1–Aug 31) occupancy probabilities and changes in occupancy over time for 12 North American bat species. This serves as one of multiple lines of evidence that inform the status and trends of bat populations. • We analyzed data from a total of 12 bat species (Table 1), 11 of which have tested positive for Pseudogymnoascus destructans (Pd), a fungal pathogen that causes white-nose syndrome (WNS)—a disease that has led to significant rates of mortality for subterranean hibernating bat species in North America. A twelfth species was also selected because of high rates of mortality at wind energy facilities. Additional species were considered but not selected due to data limitations. • We estimated occupancy probabilities for 2010 through 2019 for three species ( Myotis lucifugus , MYLU; Myotis septentrionalis , MYSE; and P erimyotis subflavus , PESU). For an additional nine species, we estimated occupancy probabilities for 2016 through 2019 ( Myotis evotis , MYEV; Myotis grisescens , MYGR; Myotis leibii , MYLE; Myotis thysanodes , MYTH; Myotis volans , MYVO; Myotis yumanensis , MYYU; Eptesicus fuscus , EPFU; Lasionycteris noctivagans , LANO; and Lasiurus cinereus , LACI). • For each species, we provide range-wide occupancy probability predictions (e.g., predicted summer occupancy distribution maps) each year at a spatial resolution of 100 km2 and provide regional estimates of mean occupancy probability aggregated at larger spatial scales (state/province/territory, range-wide). • For each species, we also provide trends over time (average annual change rate and total change rate) in mean occupancy probabilities at multiple spatial scales (state/province/territory, range-wide) and when possible, over multiple timescales (short, medium, long). • Results suggest that over the short-term (2016-2019), two ( Myotis lucifugus and Perimyotis subflavus ) of 12 species have experienced declines in range-wide average occupancy probability with at least 95% certainty. Seven species showed either minor increases or decreases in range-wide average occupancy probability but with less than 95% certainty in both trend indicators. Results over the longer term (eight years and 10 years of sampling) suggest that three hibernating species known to be highly affected by white-nose syndrome ( Myotis lucifugus , Myotis septentrionalis , and Perimyotis subflavus ) have experienced marked declines in range-wide average occupancy probabilities, with severity varying by species and region. Finally, the results for three species ( Eptesicus fuscus , Lasiurus cinereus , Lasionycteris noctivagans ) were inconclusive due to 1) borderline convergence issues in the model fitting procedure which suggests potentially unreliable estimates, 2) failure to reliably distinguish between false positives and true positive detections for ambiguous detections, and 3) largely uninformative covariates for occupancy and detection. • For Myotis lucifugus , Myotis septentrionalis , and Perimyotis subflavus we found meaningful associations in space and time between declining winter populations (likely a result of WNS) and summer occupancy distributions. • The representativeness of sampling data for each species’ status and trend estimates (e.g., state/province/territory) were also evaluated based on the percent of grid cells sampled each year with a goal of understanding the reliability of regional estimates and improving future monitoring efforts. • This work represents the most comprehensive effort to date to model North American bat distributions across their continental ranges. Despite current limitations highlighted in the discussion, the analytical methods and resulting status and trends estimates provide the best available science on summer bat populations across North America and will continue to improve over time as monitoring data sets and analytical methods improve. • Moving forward, our occupancy analyses will continue to improve with submission of more 1) data from currently underrepresented areas (i.e., improved geographic representation), 2) manually-vetted acoustic recordings, 3) capture records, and 4) roost location and count data (summer and winter).

Report↗

Evaluation of invasions and declines of submersed aquatic macrophytes

During the past 60 yr, sightings of aquatic macrophyte species in geographic regions where they had previously not been found have occurred with increasing frequency, apparently due to both greater dispersal of the plants as a result of human activities as well as better documentation of plant distribution. Intercontinental invasions, such as Myriophyllum spicatum and Hydrilla into North America, Elodea canadensis into Europe and Elodea nuttallii, Egeria densa and Cabomba caroliniana into Japan, have generally been well documented. However, the spread of an exotic species across a continent after its initial introduction (e.g., Potamogeton crispus in North America) or the expansion of a species native to a continent into hitherto unexploited territory (e.g.,the expansion of the North American native Myriophyllum heterophyllum into New England) have received little attention. Natural declines in aquatic macrophyte communities have also received little scientific study although there are many accounts of macrophyte declines. The best-documented example comes from the marine literature where extensive declines of eelgrass (Zostera) occurred in the 1930s along the Atlantic coast due to a pathogenic marine slime mold (''wasting disease''). The aim of this workshop was to identify examples of invasions or natural declines of aquatic macrophyte species throughout the world and assess the importance of environmental factors in their control. Forty-five scientists and aquatic plant managers from ten countries participated in the workshop. Eleven of the participants contributed written evaluations of species invasions and declines in their geo-graphic region. These were distributed to registered participants prior to the meeting and served as the starting-point of workshop discussions. To address the topics raised in the working papers, the participants divided into four working groups to evaluate: 1. Environmental controls of species invasions. 2. Biotic controls of species declines. 3. Abiotic controls of species declines. 4. Impact of management practices on macrophyte invasions or declines. Each working group was asked to identify existing evidence, the need for additional evidence and management implications of their topics and then requested to discuss their findings with the entire workshop at the conclusion of discussions.

Journal of Aquatic Plant Management↗

Pseudacris triseriata (western chorus frog) and Rana sylvatica (wood frog) chytridiomycosis

The chytrid fungus Batrachochytrium dendrobatidis is a known pathogen of anuran amphibians, and has been correlated with amphibian die-offs worldwide (Daszak et. al. 1999. Emerging Infectious Diseases 5:735-748). In Colorado, B. dendrobatidis has infected Boreal toads (Bufo boreas) (Muths et. al., in review) and has been identified on museum specimens of northern leopard frogs (Rana pipiens) (Carey et. al. 1999. Develop. Comp. Immunol. 23:459-472). We report the first verified case of chytrid fungus in chorus frogs (Pseudacris triseriata) and wood frogs (Rana sylvatica) in the United States. We collected seven P. triseriata, and two adult and two juvenile R. sylvatica in the Kawuneeche Valley in Rocky Mountain National Park (RMNP) during June 2001. These animals were submitted to the National Wildlife Health Center (NWHC) as part of an amphibian health evaluation in RMNP. Chorus frogs were shipped in one container. Wood frog adults and juveniles were shipped in two separate containers. Histological examinations of all chorus frogs and 3 of 4 wood frogs were positive for chytrid fungus infection. The fourth (adult) wood frog was too decomposed for meaningful histology. Histological findings consisted of multifocally mild to diffusely severe infections of the epidermis of the ventrum and hindlimb digital skin. Chytrid thalli were confined to the thickened epidermis (hyperkeratosis), were spherical to oval, and occasional thalli contained characteristic discharge pores or zoospores (Green and Kagarise Sherman 1999. J. Herpetol 35:92-103; Fellers et al. 2001. Copeia 2001:945-953). We cannot confirm that all specimens carried the fungus at collection, because infection may have spread from one individual to all other individuals in each container during transport. Further sampling of amphibians in Kawuneeche Valley is warranted to determine the rate of infection and mortality in these populations.

Colorado↗

Investigations of a large scale eared grebe (Podiceps nigricollis) die-off at the Salton Sea, California in 1992

An estimated 150,000 Eared Grebes (Podiceps nigricollis) died at the Salton Sea between 16 December 1991 and 21 April 1992. This represented the largest documented mortality event of Eared Grebes at the time and approximately 6% of the North American population. During the die-off, grebes exhibited several uncharacteristic behaviors, such as congregating at freshwater tributaries, repeatedly gulping freshwater, preening excessively, moving onto land, and allowing close approach and/or capture. Avian cholera was diagnosed in Eared Grebes collected along the north and west shoreline of the Sea late in the die-off but not from the majority of the Eared Grebes dying along the south shore. Gross and histological examinations and diagnostic testing for viruses, bacteria, and parasites did not identify the cause of mortality in the majority of Eared Grebes examined from the south shore of the Sea. Liver concentrations of arsenic, chromium, DDE, mercury, selenium, and zinc were elevated in some Eared Grebes, but none of those contaminants exceeded known thresholds for independent lethality. Poisoning by heavy metals, organochlorine, organophosphorus, or carbamate pesticides, avian botulism, and salt were ruled out as the cause of mortality. Hypotheses for the die-off are interactive effects of contaminants, immunosuppression, a yet unidentified biotoxin or pathogen present in the Salton Sea, impairment of feather waterproofing leading to hypothermia, or a unique manifestation of avian cholera that evades laboratory detection.

California↗