MORTALITY TRENDS IN NORTHERN SEA OTTERS ( ENHYDRA LUTRIS KENYONI) COLLECTED FROM THE COASTS OF WASHINGTON AND OREGON, USA (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 US 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.

Pathogen exposure and blood chemistry in the Washington, USA population of northern sea otters (Enhydra lutris kenyoni).

Northern sea otters (Enhydra lutris kenyoni) from Washington State, United States were evaluated in 2011 to determine health status and pathogen exposure. Antibodies to Brucella spp. (10%) and influenza A virus (23%) were detected for the first time in this population in 2011. Changes in clinical pathology values (serum chemistries), exposure to pathogens, and overall health of the population over the last decade were assessed by comparing 2011 data to the data collected on this population in 2001-2002. Several serum chemistry parameters were different between study years and sexes but were not clinically significant. The odds of canine distemper virus exposure were higher for otters sampled in 2001-2002 (80%) compared to 2011 (10%); likelihood of exposure significantly increased with age. Prevalence of exposure to Sarcocystis neurona was also higher in 2001-2002 (29%) than in 2011 (0%), but because testing methods varied between study years the results were not directly comparable. Exposure to Leptospira spp. was only observed in 2001-2002. Odds of Toxoplasma gondii exposure were higher for otters sampled in 2011 (97%) than otters in 2001-2002 (58%). Substantial levels of domoic acid (n = 2) and saxitoxin (n = 2) were found in urine or fecal samples from animals sampled in 2011. No evidence of calicivirus or Coxiella burnetii exposure in the Washington population of northern sea otters was found in either 2001-2002 or 2011. Changes in exposure status from 2001-2002 to 2011 suggest that the Washington sea otter population may be dealing with new disease threats (e.g., influenza) while also increasing their susceptibility to diseases that may be highly pathogenic in naïve individuals (e.g., canine distemper).

Expansion of an exotic species and concomitant disease outbreaks: pigeon paramyxovirus in free-ranging Eurasian collared doves.

Eurasian collared doves (Streptopelia decaocto) have expanded their range across the United States since their introduction several decades ago. Recent mortality events in Eurasian collared doves in Arizona and Montana, USA, during the winter of 2009-2010 were the result of pigeon paramyxovirus (PPMV), a novel disease agent. The first instance of mortality by this emerging infectious disease in this species occurred in Florida in 2001 with subsequent disease events in 2006 and 2008. Full diagnostic necropsies were performed on carcasses from the three states. PPMV was identified by RT-PCR and virus isolation and was sequenced to the VIb genotype of avian paramyxovirus-1 (APMV). Other APMVs are common in a variety of free-ranging birds, but concern is warranted because of the potential for commingling of this species with native birds, virus evolution, and threats to domestic poultry. Improved surveillance for wildlife mortality events and efforts to prevent introduction of non-native animals could reduce the threat of introducing new pathogens.

Rapid polymerase chain reaction diagnosis of white-nose syndrome in bats.

A newly developed polymerase chain reaction (PCR)-based method to rapidly and specifically detect Geomyces destructans on the wings of infected bats from small quantities (1-2 mg) of tissue is described in the current study (methods for culturing and isolating G. destructans from bat skin are also described). The lower limits of detection for PCR were 5 fg of purified fungal DNA or 100 conidia per 2 mg of wing tissue. By using histology as the standard, the PCR had a diagnostic specificity of 100% and a diagnostic sensitivity of 96%, whereas the diagnostic sensitivity of culture techniques was only 54%. The accuracy and fast turnaround time of PCR provides field biologists with valuable information on infection status more rapidly than traditional methods, and the small amount of tissue required for the test would allow diagnosis of white-nose syndrome in live animals.

Histopathologic criteria to confirm white-nose syndrome in bats.

White-nose syndrome (WNS) is a cutaneous fungal disease of hibernating bats associated with a novel Geomyces sp. fungus. Currently, confirmation of WNS requires histopathologic examination. Invasion of living tissue distinguishes this fungal infection from those caused by conventional transmissible dermatophytes. Although fungal hyphae penetrate the connective tissue of glabrous skin and muzzle, there is typically no cellular inflammatory response in hibernating bats. Preferred tissue samples to diagnose this fungal infection are rostral muzzle with nose and wing membrane fixed in 10% neutral buffered formalin. To optimize detection, the muzzle is trimmed longitudinally, the wing membrane is rolled, and multiple cross-sections are embedded to increase the surface area examined. Periodic acid-Schiff stain is essential to discriminate the nonpigmented fungal hyphae and conidia. Fungal hyphae form cup-like epidermal erosions and ulcers in the wing membrane and pinna with involvement of underlying connective tissue. In addition, fungal hyphae are present in hair follicles and in sebaceous and apocrine glands of the muzzle with invasion of tissue surrounding adnexa. Fungal hyphae in tissues are branching and septate, but the diameter and shape of the hyphae may vary from parallel walls measuring 2 microm in diameter to irregular walls measuring 3-5 microm in diameter. When present on short aerial hyphae, curved conidia are approximately 2.5 microm wide and 7.5 microm in curved length. Conidia have a more deeply basophilic center, and one or both ends are usually blunt. Although WNS is a disease of hibernating bats, severe wing damage due to fungal hyphae may be seen in bats that have recently emerged from hibernation. These recently emerged bats also have a robust suppurative inflammatory response.

Postmortem evaluation of reintroduced migratory whooping cranes in eastern North America.

Reintroduction of endangered Whooping Cranes (Grus americana) in eastern North America has successfully established a migratory population between Wisconsin and Florida. Eighty birds (47 males, 33 females) were released between 2001 and 2006, and all birds were tracked following release with satellite and/or VHF monitoring devices. By the end of 2006, 17 deaths (12 males, five females) were recorded from this population. Postmortem findings and field data were evaluated for each bird to determine the cause of death. Causes included predation (n=8, 47%), trauma (n=2, 12%), and degenerative disease (n=1, 6%); the cause of death was undetermined for 35% (n=6) of the birds. Based on physical evidence, the primary predator of the birds was the bobcat (Lynx rufus). Limited roosting habitat availability or bird behavior were likely prime factors in the occurrence of predation. Traumatic injuries and mortality were caused by gunshot, electrical utility lines, and an unknown source. The lone case of degenerative disease was due to chronic exertional myopathy associated with translocation. Available postmortem testing did not indicate the presence of infectious disease in this limited sample.

Estimating cause-specific mortality rates using recovered carcasses.

Stranding networks, in which carcasses are recovered and sent to diagnostic laboratories for necropsy and determination of cause of death, have been developed to monitor the health of marine mammal and bird populations. These programs typically accumulate comprehensive, long-term datasets on causes of death that can be used to identify important sources of mortality or changes in mortality patterns that lead to management actions. However, the utility of these data in determining cause-specific mortality rates has not been explored. We present a maximum likelihood-based approach that partitions total mortality rate, estimated by independent sources, into cause-specific mortality rates. We also demonstrate how variance estimates are derived for these rates. We present examples of the method using mortality data for California sea otters (Enhydra lutris nereis) and Florida manatees (Trichechus manatus latirostris).

Chlorinated, brominated, and perfluorinated compounds, polycyclic aromatic hydrocarbons and trace elements in livers of sea otters from California, Washington, and Alaska (USA), and Kamchatka (Russia).

Concentrations of organochlorine pesticides (DDTs, HCHs, and chlordanes), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), perfluorinated compounds (PFCs), and 20 trace elements were determined in livers of 3- to 5-year old stranded sea otters collected from the coastal waters of California, Washington, and Alaska (USA) and from Kamchatka (Russia). Concentrations of organochlorine pesticides, PCBs, and PBDEs were high in sea otters collected from the California coast. Concentrations of DDTs were 10-fold higher in California sea otters than in otters from other locations; PCB concentrations were 5-fold higher, and PBDE concentrations were 2-fold higher, in California sea otters than in otters from other locations. Concentrations of PAHs were higher in sea otters from Prince William Sound than in sea otters from other locations. Concentrations of several trace elements were elevated in sea otters collected from California and Prince William Sound. Elevated concentrations of Mn and Zn in sea otters from California and Prince William Sound were indicative of oxidative stress-related injuries in these two populations. Concentrations of all of the target compounds, including trace elements, that were analyzed in sea otters from Kamchatka were lower than those found from the US coastal locations.

Contamination status and accumulation profiles of organotins in sea otters (Enhydra lutris) found dead along the coasts of California, Washington, Alaska (USA), and Kamchatka (Russia).

Organotin compounds (OTs) including mono- to tri-butyltins, -phenyltins, and -octyltins were determined in the liver of adult sea otters (Enhydra lutris) found dead along the coasts of California, Washington, and Alaska in the USA and Kamchatka, Russia. Total concentrations of OTs in sea otters from California ranged from 34 to 4100ng/g on a wet weight basis. The order of concentrations of OTs in sea otters was total butyltins>>total octyltins> or = total phenyltins. Elevated concentrations of butyltins (BTs) were found in some otters classified under 'infectious-disease' mortality category. Concentrations of BTs in few of these otters were close to or above the threshold levels for adverse health effects. Total butyltin concentrations decreased significantly in the livers of California sea otters since the 1990s. Based on the concentrations of organotins in sea otters collected from 1992 to 2002, the half-lives of tributyltin and total butyltins in sea otters were estimated to be approximately three years.

A comparative analysis of polybrominated diphenyl ethers and polychlorinated biphenyls in Southern sea otters that died of infectious diseases and noninfectious causes.

Southern sea otters (Enhydra lutris nereis) from the California coast continue to exhibit a slower population regrowth rate than the population in Alaska. Infectious diseases have been identified as a frequent cause of death. Infectious diseases caused by varied pathogens including bacteria, fungi, and parasites were suggestive of compromised immunological health of mature animals in this population. To test the hypothesis that elevated exposure to immunotoxic contaminants such as polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) contribute to disease susceptibility via immunosuppression, we determined concentrations of PBDEs and PCBs in livers of 80 adult female sea otters that died of infectious diseases, noninfectious causes, or emaciation. Concentrations of PBDEs and PCBs in sea otter livers varied widely (10-26,800 ng/g and 81-210,000 ng/g, lipid weight, respectively). Concentrations of PBDEs in sea otters were some of the highest values reported for marine mammals so far. Although PCB concentrations in sea otters have declined during 1992-2002, the mean concentration was at the threshold at which adverse health effects are elicited. Concentrations of PBDEs and PCBs were significantly correlated, suggesting co-exposure of these contaminants in sea otters. No significant association was found between the concentrations of PBDEs and the health status of sea otters. Concentrations of PCBs were significantly higher in otters in the infectious disease category than in the noninfectious category, suggesting an association between elevated PCB concentrations and infectious diseases in Southern sea otters.

Association between perfluorinated compounds and pathological conditions in southern sea otters.

Concentrations of four perfluorinated contaminants, including perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA), were measured in liver tissue from 80 adult female sea otters collected from the California coast during 1992-2002. Concentrations of PFOS and PFOA were in the ranges of <1-884 and <5-147 ng/g, wet wt, respectively. Concentrations of PFOA in the livers of these sea otters were among the highest values reported for marine mammals to date. Liver tissue from 6 male sea otters also was analyzed and contained significantly higher concentrations of PFOS than did tissues from female otters. To examine the association between exposures and potential effects, concentrations of PFOS and PFOA were compared among the adult female otters that died from infectious diseases, noninfectious causes, and from apparent emaciation. Concentrations of both PFOA and PFOS were significantly higher in sea otters in the infectious disease category than in the noninfectious category. Concentrations of PFOS and PFOA were not significantly different between noninfectious and emaciated otters, suggesting that the poor nutritive (body) status of emaciated otters did not affectthe concentrations of perfluorochemicals in livers. Concentrations of PFOA increased significantly from 1992 to 2002, whereas PFOS concentrations increased from 1992to 1998 and then decreased after 2000. Significant association between infectious diseases and elevated concentrations of PFOS/PFOA in the livers of sea otters is a cause for concern and suggests the need for further studies.

Comparison of trace element concentrations in livers of diseased, emaciated and non-diseased southern sea otters from the California coast.

Infectious diseases have been implicated as a cause of high rates of adult mortality in southern sea otters. Exposure to environmental contaminants can compromise the immuno-competence of animals, predisposing them to infectious diseases. In addition to organic pollutants, certain trace elements can modulate the immune system in marine mammals. Nevertheless, reports of occurrence of trace elements, including toxic heavy metals, in sea otters are not available. In this study, concentrations of 20 trace elements (V, Cr, Mn, Co, Cu, Zn, Rb, Sr, Mo, Ag, Cd, In, Sn, Sb, Cs, Ba, Hg, Tl, Pb, and Bi) were measured in livers of southern sea otters found dead along the central California coast (n=80) from 1992 to 2002. Hepatic concentrations of trace elements were compared among sea otters that died from infectious diseases (n=27), those that died from non-infectious causes (n=26), and otters that died in emaciated condition with no evidence of another cause of death (n=27). Concentrations of essential elements in sea otters varied within an order of magnitude, whereas concentrations of non-essential elements varied by two to five orders of magnitude. Hepatic concentrations of Cu and Cd were 10- to 100-fold higher in the sea otters in this study than concentrations reported for any other marine mammal species. Concentrations of Mn, Co, Zn, and Cd were elevated in the diseased and emaciated sea otters relative to the non-diseased sea otters. Elevated concentrations of essential elements such as Mn, Zn, and Co in the diseased/emaciated sea otters suggest that induction of synthesis of metallothionein and superoxide dismutase (SOD) enzyme is occurring in these animals, as a means of protecting the cells from oxidative stress-related injuries. Trace element profiles in diseased and emaciated sea otters suggest that oxidative stress mediates the perturbation of essential-element concentrations. Elevated concentrations of toxic metals such as Cd, in addition to several other organic pollutants, may contribute to oxidative stress-meditated effects in sea otters.

Attempts to identify the source of avian vacuolar myelinopathy for waterbirds.

Attempts were made to reproduce avian vacuolar myelinopathy (AVM) in a number of test animals in order to determine the source of the causative agent for birds and to find a suitable animal model for future studies. Submerged vegetation, plankton, invertebrates, forage fish, and sediments were collected from three lakes with ongoing outbreaks of AVM and fed to American coots (Fulica americana), mallard ducks and ducklings (Anas platyrhynchos), quail (Coturnix japonica), and laboratory mice either via gavage or ad libitum. Tissues from AVM-affected coots with brain lesions were fed to ducklings, kestrels (Falco sparverius), and American crows (Corvus brachyrhynchos). Two mallards that ingested one sample of Hydrilla verticillata along with any biotic or abiotic material associated with its external surface developed brain lesions consistent with AVM, although neither of the ducks had clinical signs of disease. Ingestion of numerous other samples of Hydrilla from the AVM affected lakes and a lake with no prior history of AVM, other materials (sediments, algae, fish, invertebrates, and water from affected lakes), or tissues from AVM-affected birds did not produce either clinical signs or brain lesions in any of the other test animals in our studies. These results suggest that waterbirds are most likely exposed to the causative agent of AVM while feeding on aquatic vegetation, but we do not believe the vegetation itself is the agent. We hypothesize that the causative agent of AVM might either be accumulated by aquatic vegetation, such as Hydrilla, or associated with biotic or abiotic material on its external surfaces. In support of that hypothesis, two coots that ingested Hydrilla sampled from a lake with an ongoing AVM outbreak in wild birds developed neurologic signs within 9 days (ataxia, limb weakness, and incoordination), and one of two coots that ingested Hydrilla collected from the same site 13 days later became sick and died within 38 days. None of these three sick coots had definitive brain lesions consistent with AVM by light microscopy, but they had no gross or histologic lesions in other tissues. It is unclear if these birds died of AVM. Perhaps they did not ingest a dose sufficient to produce brain lesions or the lesions were ultrastructural. Alternatively, it is possible that a separate neurotoxic agent is responsible for the morbidity and mortality observed in these coots.

Exertional myopathy in whooping cranes (Grus americana) with prognostic guidelines.

Exertional myopathy developed in three whooping cranes (Grus americana) secondary to routine capture, handling, and trauma. Presumptive diagnosis of exertional myopathy was based on history of recent capture or trauma, clinical signs, and elevation of aspartate aminotransferase, alanine aminotransferase, creatine kinase, lactate dehydrogenase, and serum potassium. Treatments were attempted in each case, but ultimately were not successful. Gross and microscopic lesions at necropsy confirmed the diagnosis in each case, with the leg musculature most severely affected. Guidelines for determining prognosis of exertional myopathy in cranes have been included based on the analysis of these cases and others in the literature. As treatment is largely unrewarding, prevention remains the key in controlling exertional myopathy. Identification of predisposing factors and proper handling, immobilization, and transportation techniques can help prevent development of exertional myopathy in cranes.

Profiles of polychlorinated biphenyl congeners, organochlorine pesticides, and butyltins in southern sea otters and their prey.

Concentrations of organochlorine pesticides, polychlorinated biphenyl (PCB) congeners, and butyltins were measured in sea otters and selected prey species (invertebrates) collected from the California (USA) coast. Polychlorinated biphenyls, DDTs (sum of p,p'-dichlorodiphenyldichloroethylene [p,p'-DDE], p,p'-dichlorodiphenyldichloroethane [p,p'-DDD], and p,p'-DDT), and butyltins were the major contaminants found in sea otters and their prey. Lipid-normalized concentrations of PCBs and DDT in sea otter livers were 60- and 240-fold greater than those found in the prey. Great biomagnification of PCBs and DDT in sea otters is suggested to result from their high per-capita intake of diet compared with those of other marine mammals. Profiles of PCB congeners in sea otters and prey species suggest a great capacity of sea otters to biotransform lower-chlorinated congeners. Sea otters seem to possess a greater ability than cetaceans to metabolize PCBs. The 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents of non- and mono-ortho PCBs in sea otters and certain prey species were at or above the theoretical threshold for toxic effects.

Failure to transmit avian vacuolar myelinopathy to mallard ducks.

Avian vacuolar myelinopathy (AVM) is a neurologic disease that has been diagnosed in free-ranging birds in the southeastern United States. Bald eagles (Haliaeetus leuocephalus), American coots (Fulica americana), and mallards (Anas platyrhynchos) have been affected. Previous investigations have not determined the etiology of this disease. In November and December 2002, we attempted to induce AVM in game-farmed mallards through four, 7-day exposure trials. Mallards were housed in six groups of eight, with two of these groups serving as controls. One group was housed with AVM-affected coots; one group was tube fed daily with water from the lake where affected coots were captured; one group was tube fed daily with aquatic vegetation (Hydrilla verticillata) from the same lake; and another group was tube fed daily with sediment from the lake. No ducks exhibited clinical neurologic abnormalities consistent with AVM and no evidence of AVM was present at histopathologic examination of brain tissue. Although limitations in sample size, quantity of individual doses, frequency of dose administration, duration of exposure, and timing of these trials restrict the interpretation of the findings, AVM was not readily transmitted by direct contact, water, hydrilla, or sediment in this investigation.

Vacuolar myelinopathy in waterfowl from a North Carolina impoundment.

Vacuolar myelinopathy was confirmed by light and electron microscopic examination of mallards (Anas platyrhynchos), ring-necked ducks (Aythya collaris), and buffleheads (Bucephala albeola) collected during an epizootic at Lake Surf in central North Carolina (USA) between November 1998 and February 1999. Clinical signs of affected birds were consistent with central nervous system impairment of motor function (incoordination, abnormal movement and posture, weakness, paralysis). This is the first report of this disease in wild waterfowl (Anseriformes).

Clinical features of avian vacuolar myelinopathy in American coots.

OBJECTIVE: To characterize clinical features of avian vacuolar myelinopathy (AVM) in American coots. DESIGN: Case-control study. ANIMALS: 26 AVM-affected American coots and 12 unaffected coots. PROCEDURES: Complete physical, neurologic, hematologic, and plasma biochemical evaluations were performed. Affected coots received supportive care. All coots died or were euthanatized, and AVM status was confirmed via histopathologic findings. RESULTS: 3 severely affected coots were euthanatized immediately after examination. Seventeen affected coots were found dead within 7 days of admission, but 5 affected coots survived > 21 days and had signs of clinical recovery. Abnormal physical examination findings appeared to be related to general debilitation. Ataxia (88%), decreased withdrawal reflexes (88%), proprioceptive deficits (81%), decreased vent responses (69%), beak or tongue weakness (42%), and head tremors (31%), as well as absent pupillary light responses (46%), anisocoria (15%), apparent blindness (4%), nystagmus (4%), and strabismus (4%) were detected. Few gross abnormalities were detected at necropsy, but histologically, all AVM-affected coots had severe vacuolation of white matter of the brain. None of the control coots had vacuolation. CONCLUSIONS AND CLINICAL RELEVANCE: Although there was considerable variability in form and severity of clinical neurologic abnormalities, clinical signs common in AVM-affected birds were identified. Clinical recovery of some AVM-affected coots can occur when supportive care is administered. Until the etiology is identified, caution should be exercised when rehabilitating and releasing coots thought to be affected by AVM.

Fatal spirochetosis due to a relapsing fever-like Borrelia sp. in a northern spotted owl.

Acute septicemic spirochetosis was diagnosed in an adult male northern spotted owl (Strix occidentalis caurina) found dead in Kittitas County, Washington, USA. Gross necropsy findings included marked enlargement of the liver and spleen and serofibrinous deposits on the serous membranes lining the body cavities and the pericardial and perihepatic sacs. Microscopic observations included macrophage infiltration in the liver and spleen with mild thrombosis and multifocal necrosis, as well as hemorrhage and acute inflammation in the choroid plexus of the brain. No viruses or pathogenic bacteria were isolated from brain, liver, or spleen, and no parasites were found in blood smears or impression smears of the liver. Chlamydial culture attempts were unsuccessful and no chlamydial antibodies were detected in serum. In silver-stained microscopic sections and by transmission electron microscopy of liver, numerous long, thin, spiral-shaped bacteria were seen in the liver, spleen, cerebral ventricles, and within blood vessels in many organs. The organism was identified as a member of the Borrelia genus by sequence analysis of the PCR-amplified 16S rRNA gene. The most closely related species is B. hermsii, an agent of relapsing fever in humans in the western United States. This is the first report of a relapsing fever-related Borrelia in a wild bird.

Epizootiologic studies of avian vacuolar myelinopathy in waterbirds.

Epizootic avian vacuolar myelinopathy (AVM) was first recognized as a neurologic disease in bald eagles (Haliaeetus leucocephalus) and American coots (Fulica americana) in Arkansas, USA in 1994 and 1996, respectively, but attempts to identify the etiology of the disease have been unsuccessful to date. Between 1998 and 2001, wing clipped sentinel birds (wild American coots and game farm mallards [Anas platyrhynchos]) were released at Lake Surf, North Carolina, a lake with recurrent outbreaks of AVM, in order to gain a better understanding of the epizootiology of the disease. As early as 5-7 days post-release, sentinel coots and mallards showed neurologic signs of disease and were confirmed with AVM upon histologic examination of their brains. Serial releases of sentinel mallards during the summer, fall, and winter of 2000-01 demonstrated that exposure to the causative agent at a threshold sufficient to manifest disease was seasonal and occurred over about a 2 mo period, during November and December. Our findings that disease onset can be very rapid (5-7 days) and that exposure to the causative agent of AVM is site-specific, seasonal (late fall to early winter), and occurs over a relatively short duration (several months) supports the hypothesis that the disease is caused by a chemical substance, most likely of natural origin.