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Characterization of Pasteurella multocida isolates from wetland ecosystems during 1996 to 1999

We cultured 126 Pasteurella multocida isolates, 92 from water and 34 from sediment samples collected from wetlands in the Pacific and Central flyways of the United States between 1996 and 1999. Most (121) of the isolates were P. multocida serotype 1, but serotypes 3, 3/4, 10, and 11 were also found. Many (82) of the isolates were further characterized by DNA fingerprinting procedures and tested in Pekin ducks for virulence. Almost all the serotype 1 isolates we tested caused mortality in Pekin ducks. Serotype 1 isolates varied in virulence, but the most consistent pattern was higher mortality in male ducks than in females. We found no evidence that isolates found in sediment vs. water, between Pacific and Central flyways, or during El Nino years had consistently different virulence. We also found a number of non-serotype 1 isolates that were avirulent in Pekin ducks. Isolates had DNA fingerprint profiles similar to those found in birds that died during avian cholera outbreaks.

Journal of Wildlife Diseases

Mortality in Laysan ducks (Anas laysanensis) by emaciation complicated by Echinuria uncinata on Laysan Island, Hawaii, 1993

In November 1993, unusual mortality occurred among endangered Laysan ducks on Laysan Island, one of the remote refugia of the Northwestern Hawaiian Islands National Wildlife Refuge (USA). Ten live ducks were emaciated, and blood samples documented anemia, heterophilia, and eosinophilia. Pathology in 13 duck carcasses revealed emaciation, marked thickening of the proventricular wall, abundant mucus, and nodules in the gastrointestinal tract. Histology revealed granulomata associated with nematodes in the proventriculus, small intestines, and body walls of nine of 10 ducks examined on histology. We suspect that low rainfall and low food abundance that year contributed to enhanced pathogenicity of parasite infection, either through increased exposure or decreased host resistance. Because the Laysan duck is found only on Laysan island and is critically endangered, translocation of this species to other islands is being considered. Given that we have not seen pathology associated with Echinuria spp. in native waterfowl on other Hawaiian Islands and given the parasite's potential to cause significant lesions in Laysan ducks, it will be important to prevent the translocation of Echinuria spp.

Hawai'i

Levels of fecal corticosterone in sandhill cranes during a human-led migration

Fourteen captive-reared greater sandhill cranes ( Grus canadensis tabida ) were conditioned to follow ultralight aircraft to promote migration between Wisconsin and Florida (USA) after release. Fecal samples were collected throughout the training period in Wisconsin and during a 1,977-km human-led migration to Florida to determine fecal corticosterone (FC) concentrations by radioimmunoassay. The mean (±SE) FC concentration during the training period was 109.5±7.5 ng/g and was representative of baseline levels recorded previously from sandhill cranes. Fecal corticosterone concentrations increased in early migration compared to concentrations 1 mo prior to departure ( P <0.01) but were not different from baseline concentrations at the end of the 6-wk migration period. The variability of FC concentrations in individual samples was greater throughout the migration than the training period. Increases in FC during migration were modest and generally consistent with normal corticosterone elevations observed in migrating birds.

Florida, Wisconsin

Are wetlands the reservoir for avian cholera?

Wetlands have long been suspected to be an important reservoir for Pasteurella multocida and therefore the likely source of avian cholera outbreaks. During the fall of 1995a??98 we collected sediment and water samples from 44 wetlands where avian cholera epizootics occurred the previous winter or spring. We attempted to isolate P. multocida in sediment and surface water samples from 10 locations distributed throughout each wetland. We were not able to isolate P. multocida from any of the 440 water and 440 sediment samples collected from these wetlands. In contrast, during other investigations of avian cholera we isolated P. multocida from 20 of 44 wetlands, including 7% of the water and 4.5% of the sediment samples collected during or shortly following epizootic events. Our results indicate that wetlands are an unlikely reservoir for the bacteria that causes avian cholera.

California, Minnesota, Missouri, Nebraska, Nevada,

Peracute sodium toxicity in free-ranging black-bellied whistling duck ducklings

From 23 to 25 July 2002, 98–103 newly hatched black-bellied whistling ducks ( Dendrocygna autumnalis ) were observed alive at an inland saline lake (La Sal Vieja) in Willacy County, Texas (USA). Seventy-one (71%) died after showing signs indicative of sodium toxicity within 5 hr of entering the water; some died within minutes. Six carcasses were sent to the United States Geological Survey, National Wildlife Health Center (Madison, Wisconsin, USA) for analysis, and brain sodium levels of all ducklings were above 2,000 parts per million wet weight. More black-bellied whistling duck ducklings are likely to have been affected, but they were not observed after hatching.

Texas

Detection of Clostridium botulinum type C cells in the gastrointestinal tracts of Mozambique tilapia (Oreochromis mossambicus) by polymerase chain reaction

We established a method of directly detecting Clostridium botulinum type C cells, while minimizing spore detection, in the intestinal contents of Mozambique tilapia ( Oreochromis mossambicus ). This technique involved extraction of predominantly cellular DNA from tilapia intestinal tracts and used a polymerase chain reaction assay to detect presence of type C 1 toxin gene. We consistently detected C. botulinum type C cells in tilapia gastrointestinal contents at a level of 7.5×10 4 cells per 0.25 g material or 1.9×10 3 cells. This technique is useful for determining prevalence of the potentially active organisms within a given population of fish and may be adapted to other types of C. botulinum and vertebrate populations as well.

California

Zinc toxicosis in a free-flying trumpeter swan (Cygnus buccinator)

A trumpeter swan (Cygnus buccinator) was observed near it mill pond in Picher, Oklahoma. USA. It became weakened and emaciated after about 1 mo, was captured with little resistance, and taken into captivity for medical care. Serum chemistry results were consistent with hepatic, renal, and muscular damage. Serum zinc concentration was elevated at 11.2 parts per million (ppm). The swan was treated for suspected heavy-metal poisoning, but died overnight. Gross postmortem findings were emaciation and pectoral muscle atrophy. Histopathologic lesions in the pancreas included mild diffuse disruption of acinar architecture, severe diffuse depletion or absence of zymogen granules, occasional apoptotic bodies ics in acinar epithelial cells, and mild interstitial and capsular fibrosis. Zinc concentration in pancreas was 3,200 ppm wet weight, and was similar to that reported in the pancreases of waterfowl known to be killed by zinc toxicity. Zinc concentrations in liver (154 ppm) and kidneys (145 ppm) also were elevated. Acute tubular necrosis of the collecting tubules of the kidneys was also possibly due to zinc toxicity. To the authors' knowledge, this is the first confirmed case of zinc poisoning in a trumpeter swan associated with mining wastes..

Journal of Wildlife Diseases

Organochlorine pesticides and thiamine in eggs of largemouth bass and American alligators and their relationship with early life-stage mortality

Thiamine deficiency has been linked to early mortality syndrome in salmonids in the Great Lakes. This study was conducted to compare thiamine concentrations in American alligators ( Alligator mississippiensis ) and Florida largemouth bass ( Micropterus salmoides floridanus ) eggs from sites with high embryo mortality and high exposure to organochlorine pesticides (OCPs) (Lakes Apopka and Griffin, and Emeralda Marsh, Florida, USA) to those from sites that have historically exhibited low embryo mortality and low OCPs (Lakes Woodruff and Orange, Florida). During June-July 2000, 20 alligator clutches were collected from these sites, artificially incubated, and monitored for embryo mortality. Thiamine and OCPs were measured in one egg/clutch. During February 2002, 10 adult female bass were collected from Emeralda Marsh and Lake Woodruff and mature ovaries analyzed for thiamine and OCP concentrations. Although ovaries from the Emeralda Marsh bass contained almost 1,000-fold more OCPs compared with the reference site, Lake Woodruff, there were no differences in thiamine concentrations between sites (11,710 vs. 11,857 pmol/g). In contrast, alligator eggs from the reference site had five times the amount of thiamine compared with the contaminated sites (3,123 vs. 617 pmol/g). Similarly, clutches with > 55% hatch rates had significantly higher concentrations of thiamine compared with clutches with <54% hatch rates (1,119 vs. 201 pmol/g). These results suggest that thiamine deficiency might be playing an important role in alligator embryo survival but not in reproductive failure and recruitment of largemouth bass. The cause(s) of this thiamine deficiency are unknown but might be related to differences in the nutritional value of prey items across the sites studied and/or to the presence of high concentration of contaminants in eggs.

Florida

Anesthesia and blood sampling of wild big brown bats (Eptesicus fuscus) with an assessment of impacts on survival

We anesthetized and blood sampled wild big brown bats ( Eptesicus fuscus ) in Fort Collins, Colorado (USA) in 2001 and 2002 and assessed effects on survival. Inhalant anesthesia was delivered into a specially designed restraint and inhalation capsule that minimized handling and bite exposures. Bats were immobilized an average of 9.1±5.1 (SD) min (range 1–71, n =876); blood sample volumes averaged 58±12 μl (range 13–126, n =718). We randomly selected control (subject to multiple procedures before release) and treatment (control procedures plus inhalant anesthesia and 1% of body weight blood sampling) groups in 2002 to assess treatment effects on daily survival over a 14-day period for adult female and volant juvenile bats captured at maternity roosts in buildings. We monitored survival after release using passive integrated transponder tag detection hoops placed at openings to selected roosts. Annual return rates of bats sampled in 2001 were used to assess long-term outcomes. Comparison of 14-day maximum-likelihood daily survival estimates from control (86 adult females, 92 volant juveniles) and treated bats (187 adult females, 87 volant juveniles) indicated no adverse effect from anesthesia and blood sampling (juveniles: χ 2 =22.22, df=27, P >0.05; adults: χ 2 =9.72, df=18, P >0.05). One-year return rates were similar among adult female controls (81%, n =72, 95% confidence interval [CI] =70–91%), females treated once (82%, n =276, 95% CI=81–84%), and females treated twice (84%, n =50, 95% CI=74–94%). Lack of an effect was also noted in 1-yr return rates of juvenile female controls (55%, n =29, 95% CI=37–73%), juveniles treated once (66%, n =113, 95% CI=58–75%), and juveniles treated twice (71%, n =17, 95% CI=49–92%). These data suggest that anesthesia and blood sampling for health monitoring did not measurably affect survival of adult female and volant juvenile big brown bats.

Journal of Wildlife Diseases

Efficacy of immobilizing free-ranging elk with Telazol® and xylazine hydrochloride using transmitter-equipped darts

From January 1999 to April 2002, 14 free-ranging elk were darted with a mixture of Telazol &reg; reconstituted with xylazine hydrochloride (HCl) in a forested habitat in southwestern Oklahoma and north-central Arkansas. Elk were darted from ground blinds, tree stands, or a vehicle at distances of 14&ndash;46 m and were recovered 37&ndash;274 m from the dart site. Elk were located using radiotelemetry with 3-cc disposable Pneu-dart &reg; transmitter darts. Mean&plusmn;SD dose of Telazol &reg; and xylazine HCl was 590&plusmn;192 mg/ml and 276&plusmn;153 mg/ml, respectively, and mean time to standing after injection of reversal agent was 27 min (range: 1&ndash;65 min). The combination of Telazol &reg; and xylazine HCl successfully immobilized free-ranging elk, and transmitter-equipped darts permitted successful location of sedated elk by two people in areas of dense forest cover. The dose required to sedate elk appeared to vary depending on physiology and behavior, but no drug-induced mortality occurred despite the wide variance in the doses administered. We recommend 500 mg Telazol &reg; reconstituted with 300 mg xylazine HCl as an initial dose for a &ge;200 kg elk. If needed to achieve full sedation, up to 3 additional ml of the mixture may be administered without adverse effects.

Journal of Wildlife Diseases

Naturally occurring secondary nutritional hyperparathyroidism in cattle egrets ( Bubulcus ibis ) from Central Texas

Naturally occurring secondary nutritional hyperparathyroidism is described in the nestlings of two colonies of cattle egrets ( Bubulcus ibis ) from Central Texas (Bryan and San Antonio, Texas, USA). Nestlings from a third colony (Waco, Texas, USA) were collected in a subsequent year for comparison. Birds from the first two colonies consistently had severe osteopenia and associated curving deformities and folding fractures of their long bones. These birds also had reduced bone ash, increased osteoclasia, a marked decrease in osteoblast activity, variable lengthening and shortening of the hypertrophic zone of the epiphyseal cartilage, decreased and disorganized formation of new bone, and a marked hypertrophy and hyperplasia of the parathyroid glands as compared to birds collected from the third colony. Fibrous osteodystrophy was found in all of the birds from San Antonio and Bryan. Evidence of moderate to severe calcium deficiency was also identified in 33% of the cattle egrets collected from Waco. Gut contents of affected chicks contained predominately grasshoppers and crickets; vertebrate prey items were absent from the Bryan birds. Grasshoppers and crickets collected from fields frequented by the adult egrets in 1994 had 0.12–0.28% calcium and 0.76–0.81% phosphorus. Pooled grasshoppers and crickets collected during a subsequent wet early spring averaged 0.24% calcium and 0.65% phosphorus. Although the phosphorus content of the insect prey was adequate for growth, calcium was approximately one-third the minimum calcium requirement needed for growth for other species of birds. It was postulated that cattle egrets breeding in Central Texas have expanded their range into habitat that contains less vertebrate prey, and as a result, many nestling egrets are being fed diets that contain suboptimal calcium. Therefore, in years where vertebrate prey is scarce and forage for insect prey is reduced in calcium, nestling egrets are at risk for developing secondary nutritional hyperparathyroidism.

Texas

Molecular identification of a papilloma virus from cutaneous lesions of captive and free-ranging Florida manatees

Cutaneous papillomatous lesions were biopsied from three captive Florida manatees ( Trichechus manatus latirostris ) at Homosassa Springs State Wildlife Park (HSSWP), Homosassa, Florida, USA, and from six free-ranging Florida manatees from Crystal and Homosassa rivers, Florida. Total DNA extracted from these lesions was assayed for the presence of papilloma virus genomes using the polymerase chain reaction (PCR) with primers that target the L1 capsid protein gene. The amplification generated DNA fragments 458 base pairs in length that encompassed a highly conserved domain within the L1 capsid protein and translated into identical polypeptides of 152 amino acids, suggesting the involvement of a single papilloma virus genotype. Multiple amino acid sequence and phylogenetic analyses of the L1 fragment indicated that the Florida manatee papilloma virus is a unique and quite distinct papillomavirus from other known papilloma viruses. The emergence of this new pathogen raises concerns about its potential impact on the already endangered Florida manatee.

Journal of Wildlife Diseases

Persistence of Pasteurella multocida in wetlands following avian cholera outbreaks

Avian cholera, caused by Pasteurella multocida, affects waterbirds across North America and occurs worldwide among various avian species. Once an epizootic begins, contamination of the wetland environment likely facilitates the transmission of P. multocida to susceptible birds. To evaluate the ability of P. multocida serotype-1, the most common serotype associated with avian cholera in waterfowl in western and central North America, to persist in wetlands and to identify environmental factors associated with its persistence, we collected water and sediment samples from 23 wetlands during winters and springs of 1996a??99. These samples were collected during avian cholera outbreaks and for up to 13 wk following initial sampling. We recovered P. multocida from six wetlands that were sampled following the initial outbreaks, but no P. multocida was isolated later than 7 wk after the initial outbreak sampling. We found no significant relationship between the probability of recovery of P. multocida during resampling and the abundance of the bacterium recovered during initial sampling, the substrate from which isolates were collected, isolate virulence, or water quality conditions previously suggested to be related to the abundance or survival of P. multocida. Our results indicate that wetlands are unlikely to serve as a long-term reservoir for P. multocida because the bacterium does not persist in wetlands for long time periods following avian cholera outbreaks.

Journal of Wildlife Diseases

Raptor mortality due to West Nile virus in the United States, 2002

West Nile virus (WNV) has affected many thousands of birds since it was first detected in North America in 1999, but the overall impact on wild bird populations is unknown. In mid-August 2002, wildlife rehabilitators and local wildlife officials from multiple states began reporting increasing numbers of sick and dying raptors, mostly red-tailed hawks ( Buteo jamaicensis ) and great horned owls ( Bubo virginianus) . Commonly reported clinical signs were nonspecific and included emaciation, lethargy, weakness, inability to perch, fly or stand, and nonresponse to danger. Raptor carcasses from 12 states were received, and diagnostic evaluation of 56 raptors implicated WNV infection in 40 (71%) of these cases. Histologically, nonsuppurative encephalitis and myocarditis were the salient lesions (79% and 61%, respectively). Other causes of death included lead poisoning, trauma, aspergillosis, and Salmonella spp. and Clostridium spp. infections. The reason(s) for the reported increase in raptor mortality due to WNV in 2002 compared with the previous WNV seasons is unclear, and a better understanding of the epizootiology and pathogenesis of the virus in raptor populations is needed.

Journal of Wildlife Diseases