Concentrations of 45 Per- and Polyfluoroalkyl Substances in North American River Otters (Lontra canadensis) from West Virginia, USA.

North American river otters (Lontra canadensis) are top predators in riverine ecosystems and are vulnerable to per- and polyfluoroalkyl substance (PFAS) exposure. Little is known about the magnitude of exposure and tissue distribution of PFAS in river otters. We measured 45 PFAS in various tissues of 42 river otters collected from several watersheds in the state of West Virginia, USA. The median concentrations of ∑All (sum concentration of 45 PFAS) varied among tissues in the following decreasing order: liver (931 ng/g wet weight) > bile > pancreas > lung > kidney > blood > brain > muscle. Perfluoroalkylsulfonates (PFSAs) were the predominant compounds accounting for 58-75% of the total concentrations, followed by perfluoroalkyl carboxylates (PFCAs; 21-35%). 8:2 fluorotelomer sulfonate (8:2 FTS), 10:2 FTS, and 6:2 chlorinated polyfluoroalkyl ether sulfonate were frequently found in the liver (50-90%) and bile (96-100%), whereas hexafluoropropylene oxide dimer acid (HFPO-DA) was rarely found. The hepatic concentrations of ∑All in river otters collected downstream of a fluoropolymer production facility located along the Ohio River were 2-fold higher than those in other watersheds. The median whole body burden of ∑All was calculated to be 1580 µg. PFOS and perfluorooctanoic acid (PFOA) concentrations in whole blood of some river otters exceeded the human toxicity reference values, which warrant further studies.

Updating the AIHTS Trapping Standards to Improve Animal Welfare and Capture Efficiency and Selectivity.

In 1999, after pressure from the European Union, an Agreement on International Humane Trapping Standards (AIHTS) that would result in the banning of the steel-jawed leghold traps in the European Community, Canada, and Russia was signed. The United States implemented these standards through an Agreed Minute with the European Community. Over the last two decades, scientists have criticized the AIHTS for (1) omitting species that are commonly trapped; (2) threshold levels of trap acceptance that are not representative of state-of-the-art trap technology; (3) excluding popular traps which are commonly used by trappers although they are known to cause prolonged pain and stress to captured animals; (4) inadequate coverage of capture efficiency and species selectivity (i.e., number of captures of target and non-target species) performance. Concerns about the ability of standards and test procedures to ensure animal welfare, and about the implementation of standards, have also been voiced by wildlife biologists, managers, and conservation groups. In this review, we present a synopsis of current trapping standards and test procedures, and we compare the standards to a then contemporary 1985-1993 Canadian trap research and development program. On the basis of the above-noted concerns about AIHTS, and our experience as wildlife professionals involved in the capture of mammals, we formulated the following hypotheses: (1) the list of mammal species included in the AIHTS is incomplete; (2) the AIHTS have relatively low animal welfare performance thresholds of killing trap acceptance and do not reflect state-of-the-art trapping technology; (3) the AIHTS animal welfare indicators and injuries for restraining traps are insufficient; (4) the AIHTS testing procedures are neither thorough nor transparent; (5) the AIHTS protocols for the use of certified traps are inadequate; (6) the AIHTS procedures for the handling and dispatching of animals are nonexistent; (7) the AIHTS criteria to assess trap capture efficiency and species selectivity are inappropriate. We conclude that the AIHTS do not reflect state-of-the-art trapping technology, and assessment protocols need to be updated to include trap components and sets, animal handling and dispatching, and trap visit intervals. The list of traps and species included in the standards should be updated. Finally, the concepts of capture efficiency and trap selectivity should be developed and included in the standards. Based on our review, it is clear that mammal trapping standards need to be revisited to implement state-of-the-art trapping technology and improve capture efficiency and species selectivity. We believe that a committee of international professionals consisting of wildlife biologists and veterinarians with extensive experience in the capture of mammals and animal welfare could produce new standards within 1-2 years. We propose a series of measures to fund trap testing and implement new standards.

The development and use of a spatially explicit model for river otters to evaluate environmental hazards: a case study on the Department of Energy's Savannah River Site.

The Department of Energy's (DOE) Savannah River Site (SRS) faces a legacy of radionuclide and metal contamination from industrial processes that occurred throughout the site. Northern river otters (Lontra canadensis) are appropriate receptors for studying the effects of long-term, low-level contamination because they are long-lived, higher trophic level organisms susceptible to accumulating high levels of pollutants. The purpose of this study was to use latrine surveys to examine patterns of wetland latrine usage; explicitly model northern river otter resource selection on the landscape level; and utilize the model results within an ecological risk assessment (ERA) framework to assess potential effects of metals and radiocesium (137Cs) on the population for the SRS as a case study. River drainages and associated wetlands were surveyed for latrine sites and scats were collected and analyzed for 137Cs activity to validate model results. The spatially explicit resource model predicted otter drainage reach use and was used in an ERA to develop exposure models for nine heavy metals as well as 137Cs on the SRS population of river otters. The evaluation predicted that the only contaminant occurring at high enough levels to cause population effects was mercury and that the observed concentrations were probably not high enough to cause significant impairment. However, multiple metals were above action level thresholds. The field validation process showed an unexpected preference for one man-made treatment wetland that was heavily contaminated, showing that the ERA process is complex and must be approached using multiple scales.

STRESS LEVELS IN CAPTURED RIVER OTTERS (LONTRA CANADENSIS) DECREASED AFTER TRANSPORTATION TO REINTRODUCTION SITES.

The level of stress that animals endure during capture, handling, transportation, and release processes is a major concern of animal reintroduction projects. Animals under chronic stress are more susceptible to disease and other deleterious issues that could reduce their survival in a new environment. Northern river otters ( Lontra canadensis ) have been reintroduced in 22 states in the United States and may be susceptible to developing chronic stress during the reintroduction process. We assessed stress levels in five river otters captured from wild populations in Washington, held in captivity for up to 21 days, and then transported to New Mexico for reintroduction. Glucocorticoid levels in fecal samples of all otters tested decreased from when they were held captive in Washington to the time of release. This outcome suggests that habituation to captivity before transport and release may serve to minimize the likelihood of an otter being released while experiencing a potentially burdensome level of stress.

Low Titers of Canine Distemper Virus Antibody in Wild Fishers (Martes pennanti) in the Eastern USA.

Canine distemper virus (CDV) infects species in the order Carnivora. Members of the family Mustelidae are among the species most susceptible to CDV and have a high mortality rate after infection. Assessing an animal's pathogen or disease load prior to any reintroduction project is important to help protect the animal being reintroduced, as well as the wildlife and livestock in the area of relocation. We screened 58 fishers for CDV antibody prior to their release into Pennsylvania, US, as part of a reintroduction program. Five of the 58 (9%) fishers had a weak-positive reaction for CDV antibody at a dilution of 1:16. None of the fishers exhibited any clinical sign of canine distemper while being held prior to release.

Utility of two modified-live virus canine distemper vaccines in wild-caught fishers (Martes pennanti).

BACKGROUND: Canine distemper virus (CDV) infects families in the order Carnivora. As a preventive measure, vaccinations against CDV are frequently given to mustelids in captive environments. OBJECTIVES: Our objectives were to compare the utility between two modified-live virus canine distemper vaccines (MLV CDV's), Fervac-D® (no longer manufactured) and Galaxy-D® (now manufactured by MSD Animal Health as part of a multivalent vaccine), in developing an immune response in wild-caught fishers. ANIMALS AND METHODS: The Pennsylvania Fisher Reintroduction Project (PFRP) used 14 wild-caught fishers during one year of the project to evaluate the utility of vaccinations against CDV as part of any reintroduction project. Fishers were injected subcutaneously in the nape of the neck with their designated vaccine. RESULTS: Fervac-D® did not effectively stimulate development of a serologic antibody response, whereas Galaxy-D® had adequate seroconversion or rise of titer levels to suggest that the general use of MLV CDV may be suitable in fishers pending further studies. CONCLUSION: We recommend that future studies be conducted, evaluating the use of currently produced vaccines in fishers. Future research should also focus on the length of days required between administration of primary and booster vaccines to achieve sufficient immune response. CLINICAL IMPORTANCE: If only primary doses are required, then hard-release reintroduction projects for fishers could be recommended. If primary and booster vaccines are required then soft-release reintroduction projects should be recommended that include captive management periods, allowing for appropriate vaccination intervals needed to maximize the probability of protection against CDV.

Efficacy of two canine distemper vaccines in wild Nearctic river otters (Lontra canadensis).

Canine distemper virus (CDV), a contagious morbillivirus, infects families in the order Carnivora, including Nearctic river otters (Lontra canadensis). As a preventative measure, vaccinations against CDV are frequently given to mustelids in captive environments. The Pennsylvania River Otter Reintroduction Project (PRORP) used wild-caught river otters to evaluate the efficacy and need for vaccinations against CDV as part of any reintroduction project. The objectives of this study were to: 1) evaluate the prevalence of exposure to CDV in wild river otters, 2) determine the immunologic response of river otters (i.e., seroconversion) after vaccination with a single (primary) vaccine dose compared to a second (booster) dose of Galaxy-D, a modified live-virus canine distemper (CD) vaccine (MLV CDV), and 3) determine the immunologic response after being vaccinated with a primary vaccination compared to a booster dose of Fervac-D, an MLV CDV. River otters were injected subcutaneously in the nape of the neck with their designated vaccine. Timeframes for collection of blood samples and/or injection of booster vaccines varied depending on the parameters of PRORP. Ten of the 22 river otters had positive prevaccination titer levels to CD. Both vaccines, Galaxy-D and Fervac-D, produced sufficient seroconversion or rise of titer levels (86% and 57%, respectively) to recommend the use of vaccines in wild river otters. Future studies are recommended to evaluate currently produced CD vaccines. Future research should also focus on the number of days required between administration of primary and booster vaccines to achieve sufficient immune response. If only a primary dose is required, then hard-release reintroduction projects for river otters could be recommended. If primary and booster vaccines are required then soft-release reintroduction projects should be recommended. Soft-release projects should include captive management periods that allow for appropriate vaccination intervals and boosters needed to maximize the probability of protection against CDV.

Impact of population expansion on genetic diversity and structure of river otters (Lontra canadensis) in Central North America.

Populations of North American river otters (Lontra canadensis) declined throughout large portions of the continent during the early 1900s due to habitat degradation and unregulated trapping. River otters had been extirpated in North Dakota (ND), but the Red River Valley has since been recolonized, with potential source populations including the neighboring states of Minnesota or South Dakota, or the Canadian province of Manitoba (MB). We genotyped 9 microsatellite loci in 121 samples to determine the source population of river otters in the Red River Valley of ND, as well as to assess population structure and diversity of river otters in central North America. Overall, genetic diversity was high, with an average observed heterozygosity of 0.58. Genetic differentiation was low (F ST < 0.05) between river otters in ND and those of Minnesota, suggesting that eastern ND was recolonized by river otters from Minnesota. River otters from MB were genetically distinct from all other sampled populations. Low genetic differentiation (F ST = 0.044) between South Dakota and Louisiana (LA) suggested that reintroductions using LA stock were successful. The genetic distinctiveness of river otters from different geographic regions should be considered when deciding on source populations for future translocations.

Chemical restraint of fishers (Martes pennanti) with ketamine and medetomidine-ketamine.

We chemically restrained fishers (Martes pennanti) as part of a captive-management protocol designed to facilitate veterinary evaluation and treatment, and conditioning on a high-calorie diet before reintroduction in Pennsylvania. We compared the safety and efficacy of ketamine (KET) and medetomidine-ketamine (MED-KET) by monitoring immobilization intervals (induction time, down time, alert time, and recovery time) and physiologic responses (pulse rate, respiration rate, rectal temperature, blood pressure, oxygen saturation, and mean arterial pressure) during restraint. We administered MED-KET at 0.4 mg MED combined with 20.0 mg KET to males and at 0.2 mg MED combined with 10.0 mg KET to females. The x +/- SD dosages were MED 0.07 +/- 0.008 mg/kg + KET 3.7 +/- 0.5 mg/ kg for males and MED 0.07 +/- 0.007 mg/kg + KET 3.6 +/- 0.3 mg/kg for females. KET alone was administered at 100.0 mg to males and at 50.0 mg to females. resulting in x +/- SD dosages of 18.7 +/- 1.8 mg/kg for males and 19.2 +/- 2.2 mg/kg for females. Mean induction time did not differ between fishers restrained with MED-KET (4.6 min) and KET (4.5 min). However, compared with KET, MED-KET resulted in longer mean down time (36.2 vs. 142.2 min), alert time (40.8 vs. 146.8). and recovery time (81.1 vs. 199.4 min). Fishers that received MED-KET were mildly bradycardic and hypertensive compared with those that received KET. Although KET resulted in increased muscle tension and labored respiration, it would be effective for performing brief, noninvasive procedures for fishers because induction was rapid, recovery was short and calm, anesthesia was not profound, and physiologic response was generally expected on the basis of known drug pharmacology. Medetomidine-ketamine also immobilized fishers effectively, providing rapid induction, physiologic response typical to alpha2 agonism, calm recovery, and possibly a plane of anesthesia adequate for invasive procedures such as tooth removal or surgery.

Microsatellite variation in the reintroduced Pennsylvania elk herd.

Relocation programs have restored elk (Cervus elaphus) to portions of its vast historical range. We examine the consequences of these relocation programs by assessing variation at 10 microsatellite loci in three elk herds, a source herd (Yellowstone National Park), a large herd reintroduced from Yellowstone (Custer State Park) and a bottlenecked herd reintroduced from both Yellowstone and Custer (the Pennsylvania herd). Observed single locus heterozygosities ranged from 0.000 to 0.739. Multi-locus heterozygosities ranged from 0.222 to 0.589. Although significant differences were detected among all three herds, the Yellowstone National Park and Custer State Park herds possessed similar levels of variation and heterozygosity, and the genetic distance between these two herds was small. The Pennsylvania herd, on the other hand, experienced a 61.5% decrease in heterozygosity relative to its source herds, possessed no unique and few rare alleles, and the genetic distances between the Pennsylvania herd and its sources were large. Simulations were performed to identify bottleneck scenarios in agreement with levels of variation in the Pennsylvania herd. Our data confirm that the rate of population growth post-relocation may have important genetic consequences and indicate that theoretical predictions regarding the maintenance of genetic variation during relocation events must be viewed with caution when small numbers of a polygynous species are released.