MORTALITY AND MORBIDITY REVIEW IN THE UNITED KINGDOM EX SITU POPULATION OF EUROPEAN WILDCATS (FELIS SILVESTRIS) BETWEEN 2000 AND 2021.

The Scottish population of the European wildcat (Felis silvestris), the only remaining native felid species in the United Kingdom, is critically endangered and was declared functionally extinct by the International Union for Conservation of Nature in 2019. This retrospective study investigated the causes of morbidity and mortality reported in the United Kingdom captive wildcat population from 01 January 2000 to 31 December 2021. The aim was to assess the health and welfare of ex situ wildcats and, by making recommendations for management based on study findings, contribute to the sustainability of the population under managed care. Younger wildcats accounted for 85.7% of all morbidity cases (kittens, 7/77; young adults, 59/77), and the gastrointestinal (67.5% [52/77]), musculoskeletal (10.4% [8/77]), and integumentary (5.2% [4/77]) systems were most affected. Mortality was primarily associated with disease of the gastrointestinal (13.5% [12/89]), respiratory (13.5% [12/89]), neurological (5.6% [5/89]), and renal (5.6% [5/89]) systems. One quarter of all the histopathology examinations reported gastritis with associated Helicobacter-like organisms, often combined with pancreatitis or cholangiohepatitis. Neonates represented 25% (22/89) of all deaths, a high percentage compared with that of previous reviews in other nondomestic felids.

APPLICATION OF A NOVEL ASPERGILLUS LATERAL-FLOW DEVICE IN THE DIAGNOSIS OF ASPERGILLOSIS IN CAPTIVE GENTOO PENGUINS (PYGOSCELIS PAPUA PAPUA).

Aspergillosis is the primary fungal disease affecting captive penguins globally. Its diagnosis remains challenging, and currently no tests are both sensitive and specific for the detection of early infection. The present study evaluated a recently developed Aspergillus lateral-flow device (AspLFD) for the detection of Aspergillus spp. antigen in plasma and glottis mucus from captive penguins. In a pilot retrospective study, banked frozen plasma samples from captive penguins were reviewed: samples from 11 gentoo penguins (Pygoscelis papua papua) and 4 king penguins (Aptenodytes patagonicus) fulfilled the inclusion criteria and were used in the analysis. Positive plasma AspLFD test results were found in 80% (four of five) of the aspergillosis-positive cases tested. All of the aspergillosis-negative cases tested negative (10 of 10) on the AspLFD test. In a cohort prospective study, paired plasma and glottis swab samples were opportunistically and nonrandomly collected from captive gentoo penguins. In total, 26 penguins were tested. In the negative control group, AspLFD test was negative on plasma and swab in 100% of birds (14 of 14). In the aspergillosis-positive group, AspLFD test was positive on plasma samples from 33% (4 of 12) of birds, on swab samples from 50% (6 of 12) of birds, and on either plasma or swab samples from 75% (9 of 12) of birds. The AspLFD is currently used for the diagnosis of aspergillosis in humans and also shows promise for use in penguins. Larger prospective studies are recommended.

HYPERVITAMINOSIS D IN A GIANT ANTEATER (MYRMECOPHAGA TRIDACTYLA) AND A LARGE HAIRY ARMADILLO (CHAETOPHRACTUS VILLOSUS) RECEIVING A COMMERCIAL INSECTIVORE DIET.

Hypervitaminosis D was diagnosed in a giant anteater (Myromecophaga tridactyla) and a large hairy armadillo (Chaetophractus villosus) being fed a commercial insectivore diet. Clinical findings included weight loss, reduced appetite, vomiting, and suspected abdominal discomfort. Hypercalcemia (3.68 and 2.04 mmol/L total and ionized calcium, respectively) was detected in the anteater, and plasma 25(OH)D levels were measured and found to be 808.7 and 379.4 nmol/L for the anteater and armadillo, respectively. Dietary change resulted in a reduction of 25(OH)D levels in both animals and resolution of hypercalcemia in the giant anteater. Dietary analysis of the commercial insectivore food revealed levels of vitamin D3 higher than the data-sheet values. This case report demonstrates that hypervitaminosis D in Xenarthra can be associated with significant clinical signs.

USE OF CLINDAMYCIN IN PALLAS' CATS [OTOCOLOBUS (FELIS) MANUL] TO REDUCE JUVENILE TOXOPLASMOSIS-ASSOCIATED MORTALITY RATES.

Pallas' cat [Otocolobus (Felis) manul] experiences a high mortality rate from toxoplasmosis. During the period 2006-2016, the overall mortality rate for this species from all causes during the first year of life was 71.59% in European Association of Zoos and Aquaria institutions, with the most significant infectious cause from systemic toxoplasmosis (20.6%) as confirmed by postmortem examination and histopathology. Clindamycin was used starting in 2014 in two collections that had previously experienced 100% mortality rates by toxoplasmosis in kittens less than one year of age, covering key Toxoplasma gondii exposure periods for kittens (n = 17) as a prophylactic measure. This protocol resulted in a 67.03% (95% confidence interval 41.76-78.61%) reduction in the first year mortality rate over a two-year period to 5.88% in those animals treated.

MANDIBULAR SQUAMOUS CELL CARCINOMA IN A ROCK HYRAX ( PROCAVIA CAPENSIS).

A 9-yr-old, entire female captive rock hyrax ( Procavia capensis) was presented with a 1-wk history of sialorrhea. On clinical examination, a mass was identified, encompassing the rostral mandible and intermandibular area, with associated mucosal ulceration, marked gingival recession, and loosening or loss of adjacent teeth. Skull radiography and cytology of fine-needle aspirates of the mass were suggestive of squamous cell carcinoma. Based on a suspected poor prognosis, the animal was humanely euthanized. Postmortem histological examination of samples confirmed a diagnosis of oral squamous cell carcinoma with invasion of mandibular bone. Neoplasia is uncommonly reported in hyraxes, which has led to the assumption that they may share mechanisms of cancer resistance with elephants, their closest extant relatives. This is the first report of squamous cell carcinoma in this species.

SUSPECTED HYPOVITAMINOSIS A-ASSOCIATED SALT GLAND ADENITIS IN NORTHERN ROCKHOPPER PENGUINS ( EUDYPTES MOSELEYI).

Supraorbital salt-excreting glands are present in at least 10 avian orders and are largest in marine species, including penguins. Diseases of the avian salt gland have been described infrequently. From September 2015, five captive northern rockhopper penguins ( Eudyptes moseleyi) were presented over a 6-wk period for unilateral or bilateral supraorbital swellings. In September 2016, two cases recurred and two additional cases were identified. Histopathology demonstrated salt gland adenitis with extensive squamous metaplasia. Blood plasma testing demonstrated marked vitamin A and E deficiencies within the colony. Prolonged frozen storage of feed-fish was implicated as a cause of vitamin depletion; reducing storage times and addition of dietary supplementation prevented recurrence.

CLINICAL SIGNS, ANTEMORTEM DIAGNOSTICS, AND PATHOLOGICAL FINDINGS ASSOCIATED WITH MYCOBACTERIUM AVIUM SUBSPECIES PARATUBERCULOSIS INFECTION IN MISHMI TAKIN ( BUDORCAS TAXICOLOR TAXICOLOR).

Mycobacterium avium subspecies paratuberculosis (MAP) is a cause of contagious and typically fatal enteric disease, primarily affecting ruminant and pseudoruminant species. During a MAP outbreak in a captive collection, six of nine adult Mishmi takin ( Budorcas taxicolor taxicolor) showed marked weight loss over 1-3 mo, followed by an acute deterioration. Fecal culture and microscopy failed to identify MAP shedding. Necropsy findings included grossly normal intestines and marked enlargement of mesenteric lymph nodes. Histological findings included multibacillary granulomatous enteritis, mesenteric lymphadenitis, and periportal hepatitis. MAP was confirmed by culture of intestinal and lymph node tissues from the index case. Results of antemortem serological testing using an indirect ELISA (ID SCREEN® Paratuberculosis Indirect) were corroborated by findings at necropsy or survival of the outbreak. Mishmi takin appear to show high MAP susceptibility and a rapid disease course compared with domestic ruminant species.

Development and validation of a versatile non-invasive urinary steroidomics method for wildlife biomonitoring.

Wildlife conservation is often challenged by a lack of knowledge about the reproduction biology and adaptability of endangered species. Although monitoring steroids and related molecules can increase this knowledge, the applicability of current techniques (e.g. immunoassays) is hampered by species-specific steroid metabolism and the requisite to avoid invasive sampling. This study presents a validated steroidomics method for the (un)targeted screening of a wide range of sex and stress steroids and related molecules in urine using ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). In total, 50 steroids (conjugated and non-conjugated androgens, estrogens, progestogens and glucocorticoids) and 6 prostaglandins could be uniquely detected. A total of 45 out of 56 compounds demonstrated a detection limit below 0.01 ng µL-1. Excellent linearity (R2 > 0.99), precision (CV < 20 %), and recovery (80-120 %) were observed for 46, 41, and 39 compounds, respectively. Untargeted screening of pooled giant panda and human samples yielded 9691 and 8366 features with CV < 30 %, from which 84.1 % and 83.0 %, respectively, also demonstrated excellent linearity (R2 > 0.90). The biological validity of the method was investigated on male and female giant panda urine (n = 20), as well as pooled human samples (n = 10). A total of 24 different steroids were detected with clear qualitative and quantitative differences between human and giant panda samples. Furthermore, expected differences were revealed between female giant panda samples from different reproductive phases. In contrast to traditional biomonitoring techniques, the developed steroidomics method was able to screen a wide range of compounds and provide information on the putative identities of metabolites potentially important for reproductive monitoring in giant pandas. These results illustrate the advancements steroidomics brings to the field of wildlife biomonitoring in the pursuit to better understand the biology of endangered species.

The novel use of urinary androgens to optimise detection of the fertile window in giant pandas.

Abstract: Giant pandas are mono-estrus seasonal breeders, with the breeding season typically occurring in the spring. Successful fertilization is followed by an embryonic diapause, of variable length, with birth in the late summer/autumn. There is a need for additional understanding of giant panda reproductive physiology, and the development of enhanced biomarkers for impending proestrus and peak fertility. We aimed to determine the utility of non-invasive androgen measurements in the detection of both proestrus and estrus. Urine from 20 cycles (-40 days to +10 days from peak estrus) from 5 female giant pandas was analyzed for estrogen, progestogens and androgens (via testosterone and DHEA assays), and hormone concentrations were corrected against urinary specific gravity. Across proestrus, estrogens increased while progestogens and androgens decreased - at the point of entry into proestrus, androgens (as detected by the testosterone assay) decreased prior to progestogens and gave 4 days advanced warning of proestrus. At the time of peak estrus, androgens (as detected by the DHEA assay) were significantly increased at the time of the decrease in estrogen metabolites from the peak, acting as an alternative confirmatory indicator of the fertile window. This novel finding allows for enlargement of the preparative window for captive breeding and facilitates panda management within breeding programmes. Androgens allow an enhanced monitoring of giant panda estrus, not only advancing the warning of impending proestrus, but also prospectively identifying peak fertility. Lay summary: Giant pandas have one chance at pregnancy per year. The 2-day fertile window timing varies by year and panda. This is monitored by measuring the level of estrogens in the urine, which increase, indicating an upcoming fertile period. After 1-2 weeks of increase, estrogens peak and fall, marking the optimal fertile time. We tested other hormones to see if we can predict the fertile window in advance, and the specific fertile time with more accuracy. In 20 breeding seasons from 5 females, we found androgens, usually thought of as male hormones, had an important role. Testosterone gives 4 days advanced warning of estrogens increasing. DHEA identified peak estrogen and the fertile time before needing to see a confirmed decrease in estrogen itself. Therefore, androgens help improve monitoring of the giant panda breeding season, giving early warning of fertility, key in facilitating captive breeding and giant panda conservation.

Ultrasound measurement of optic nerve sheath diameter in patients with a clinical suspicion of raised intracranial pressure.

BACKGROUND: To assess if ultrasound measurement of the optic nerve sheath diameter (ONSD) can accurately predict the presence of raised intracranial pressure (ICP) and acute pathology in patients in the emergency department. METHODS: This 3-month prospective observational study used ultrasound to measure the ONSD in adult patients who required CT from the emergency department. The mean ONSD from both eyes was measured using a 7.5 MHz ultrasound probe on closed eyelids. A mean ONSD value of >0.5 cm was taken as positive. Two radiologists independently assessed CT scans from patients in the study population for signs of raised ICP and signs of acute pathology (cerebrovascular accident, subarachnoid, subdural or extradural haemorrhage and tumour). Specificity, sensitivity and κ values, for interobserver variability between reporting radiologists, were generated for the study data. RESULTS: In all, 26 patients were enrolled into the study. The ONSD measurement was 100% specific (95% CI 79% to 100%) and 86% sensitive (95% CI 42% to 99%) for raised ICP. For any acute intracranial abnormality the value of ONSD was 100% specific (95% CI 76% to 100%) and 60% sensitive (95% CI 27% to 86%). κ Values were 0.91 (95% CIs 0.73 to 1) for identification of raised ICP on CT and 0.84 (95% CIs 0.62 to 1) for any acute pathology on CT, between the radiologists. CONCLUSIONS: This study shows that ultrasound measurement of ONSD is sensitive and specific for raised ICP in the emergency department. Further observational studies are needed but this emerging technique could be used to focus treatment in unstable patients.

Eurasian beaver (Castor fiber) health surveillance in Britain: Assessing a disjunctive reintroduced population.

BACKGROUND: Numerous translocations of Eurasian beavers have occurred with little implementation of standardised health screening. Pre-release health screening enables the selection of individuals with the best survival prospects and reduces potential health risks, but this is by-passed during unofficial releases. Beaver reintroduction to Britain has been haphazard and currently disjunctive populations of varying status exist. METHODS: This observational cross section study investigated the health status of three beaver populations, with 90 live beavers tested for a range of pathogens comprising 56 from Tayside (unofficially released Scotland), nine from Knapdale (officially released Scotland) and 25 from Devon (unofficially released England). In addition, a further 32 cadavers were screened (25 from Tayside and seven from Knapdale). RESULTS: All beavers were in good physical condition, did not harbour any non-native disease or parasites of concern and demonstrated remarkably low levels of any disease or parasite exposure. CONCLUSION: Beavers are establishing and adapting well to British landscapes and are not acting as reservoirs of significant zoonotic diseases. Official, licensed reintroduction programmes may appear overly convoluted; however, reputational damage of unofficial releases should be considered, along with the health and welfare of the animals involved and collateral damage to other wildlife, domestic animals and humans.

Evidence of Leptospira species and their significance during reintroduction of Eurasian beavers (Castorfiber) to Great Britain.

The Scottish Beaver Trial (SBT) reintroduced the Eurasian beaver (Castor fiber) in 2009 using wild-caught Norwegian beavers. This included a six-month prerelease quarantine in Devon, England. The International Union for Conservation of Nature (IUCN) and government guidelines for health screening were followed, including testing for Leptospira species. Unlicensed beavers, from Germany, were also identified in Scotland (Tayside) and Devon (later forming the River Otter Beaver Trial (ROBT)) and were health-screened under licence. Due to positive Leptospira species results and lack of prerelease screening in ROBT and Tayside, beavers from Germany and Norway (range sources) were screened. One hundred and fifty-six samples from 151 beavers were analysed by Leptospira species quantitative PCR (qPCR) (n=73 kidney (postmortem)/urine samples (antemortem)) or microscopic agglutination test (MAT, Leptospira pools 1-6) (n=83 serum samples). No beavers from Norway (95 per cent confidence interval (CI) 0-5.6 per cent, n=52), Tayside or SBT postrelease (95 per cent CI 0-4.6 per cent, n=63) tested positive. Seven beavers from Germany and Devon were positive. This gives an overall 9.3 per cent (95 per cent CI 5.2-15.1 per cent) exposure level, of which 4.6 per cent (95 per cent CI 1.9-9.3 per cent) suggested infection on a positive qPCR (n=1) or MAT titre of at least 1/400 (n=6), although none had abnormal physical, biochemical or haematological changes. This study suggests that Leptospira species infection in wild Eurasian beavers occurs at a low level, has no sex bias and does not appear to cause significant morbidity or mortality.

Urinary estrogens as a non-invasive biomarker of viable pregnancy in the giant panda (Ailuropoda melanoleuca).

Female giant pandas show complex reproductive traits, being seasonally monoestrus, displaying a variable length embryonic diapause and exhibiting pseudopregnancy. Currently, there is no confirmatory non-invasive biomarker of blastocyst implantation or pregnancy. This study aimed to monitor urinary estrogens across gestation in pregnancy (n = 4), pseudopregnancy (n = 4) and non-birth cycles (n = 5) in the giant panda. A pregnancy-specific profile of estrogens corrected for urinary specific gravity was identified during the gestation period. Pregnant females showed increasing concentrations of estrogens for 29 days until birth, no increase was observed during pseudopregnancy and the two profiles were distinguishable from each other for the final 2 weeks of the cycle suggesting the estrogens are of placental origin. This allowed a nomogram, starting at a known fixed point during the cycle, to be created and tested with cycles of known outcome, and cycles which were inseminated but did not result in a birth. Non-birth profiles showed deviations from that of pregnancy. We believe these deviations indicate the point of failure of the placenta to support a developing cub. Non-invasive longitudinal monitoring of estrogen concentrations therefore has the potential to be developed as a panda pregnancy test to predict viable cub development.

Absence of hantavirus in water voles and Eurasian beavers in Britain.

Hantaviruses are RNA viruses (order Bunyavirales, family Hantaviridae) found in rodent, bat and insectivore reservoir-hosts and have been reported as an emerging significant zoonotic risk in Europe. As part of two native semiaquatic rodent restoration projects, tissue and urine samples were tested for hantavirus from water voles (Arvicola amphibius) (n=26, in 2015) and Eurasian beavers (Castor fiber) (n=20, covering 2010-2015) using a pan-hantavirus nested real-time PCR test. Kidney and lung samples were also analysed by light microscopy after haematoxylin and eosin staining of formalin-fixed paraffin wax sections. Individuals selected included those forming the source of release animals and from those already free-living in Britain in areas targeted for release, to identify existing reservoirs. For water voles all tested individuals were from Britain (n=26); for beavers some were from Britain (Scotland) (n=9) and some were samples from wild Norwegian (Telemark region) (n=6) and German (Bavaria region) animals (n=5) that formed the source of accepted wild populations currently present in Scotland. All samples tested from both species were negative for hantavirus RNA and showed no significant histopathological changes suggesting that reservoir infection with hantavirus in water voles in Britain and Eurasian beavers present in Britain, Norway and Bavaria, Germany, is unlikely.

Outcomes of a 'One Health' Monitoring Approach to a Five-Year Beaver (Castor fiber) Reintroduction Trial in Scotland.

The Scottish Beaver Trial, involving the translocation and release of 16 wild Norwegian beavers (Castor fiber) to Scotland, provides a good example of a 'One Health' scientific monitoring approach, with independent monitoring partners on ecology and public health feeding into veterinary health surveillance. Pathogen detection did not prohibit beaver release, although eight beavers were seropositive for Leptospira spp. Six deaths (37.5%) occurred during Rabies quarantine, followed by the death of two animals shortly after release and two wild-born kits due to suspected predation. Two host-specific parasites, the beaver fluke (Stichorchis subtriquetrus) and beaver beetle (Platypsyllus castoris) were also reintroduced.

Plasma protein electrophoresis as a prognostic indicator in Aspergillus species-infected Gentoo penguins (Pygoscelis papua papua).

BACKGROUND: Avian aspergillosis presents a significant threat to captive penguin populations. Currently, a lack of objective prognostic factors limits disease staging, objective reassessment throughout treatment, comparative evaluation of treatment regimes, and appropriate timing of euthanasia. OBJECTIVE: The study objective was to investigate absolute and relative plasma protein fractions by agarose gel electrophoresis (EPH) as predictors of survival in Gentoo penguins (Pygoscelis papua papua) under treatment for aspergillosis. METHODS: One hundred and eighty-three EPH profiles from individual clinical cases were examined retrospectively. Animal survival to 90 days post sampling was established from clinical records; birds either survived (n = 146) or died within 90 days (n = 37), and time to death was recorded. RESULTS: Fourteen variables showed statistically significant differences (P < .05) between surviving and dying birds. Receiver operating characteristic curve analysis identified total albumin concentration (albumin + prealbumin) and albumin-to-globulin (A:G) ratio as having strongest discriminatory values (95% CI) at 0.788 (0.710-0.866) and 0.784 (0.696-0.871), respectively. Albumin (concentration and percentage of total protein) displayed moderate discriminatory value but additionally a weak positive correlation with time to death (95% CI); r = .353 (0.033-0.608) and .424 (0.116-0.658), respectively. CONCLUSIONS: Optimized test cutoffs for total albumins, albumin (concentration and percentage of total protein), and A:G ratio achieved moderate sensitivity and specificity, strong negative predictive values, but weak positive predictive values due to a low prevalence of death. Selection of appropriate test cutoff values may provide valuable adjunctive prognostic tools for clinical decision-making when the prognosis is difficult to assess clinically.

Giant Panda (Ailuropoda melanoleuca) Buccal Mucosa Tissue as a Source of Multipotent Progenitor Cells.

Since the first mammal was cloned, the idea of using this technique to help endangered species has aroused considerable interest. However, several issues limit this possibility, including the relatively low success rate at every stage of the cloning process, and the dearth of usable tissues from these rare animals. iPS cells have been produced from cells from a number of rare mammalian species and this is the method of choice for strategies to improve cloning efficiency and create new gametes by directed differentiation. Nevertheless information about other stem cell/progenitor capabilities of cells from endangered species could prove important for future conservation approaches and adds to the knowledge base about cellular material that can be extremely limited. Multipotent progenitor cells, termed skin-derived precursor (SKP) cells, can be isolated directly from mammalian skin dermis, and human cheek tissue has also been shown to be a good source of SKP-like cells. Recently we showed that structures identical to SKPs termed m-SKPs could be obtained from monolayer/ two dimensional (2D) skin fibroblast cultures. Here we aimed to isolate m-SKPs from cultured cells of three endangered species; giant panda (Ailuropoda melanoleuca); red panda (Ailurus fulgens); and Asiatic lion (Panthera leo persica). m-SKP-like spheres were formed from the giant panda buccal mucosa fibroblasts; whereas dermal fibroblast (DF) cells cultured from abdominal skin of the other two species were unable to generate spheres. Under specific differentiation culture conditions giant panda spheres expressed neural, Schwann, adipogenic and osteogenic cell markers. Furthermore, these buccal mucosa derived spheres were shown to maintain expression of SKP markers: nestin, versican, fibronectin, and P75 and switch on expression of the stem cell marker ABCG2. These results demonstrate that giant panda cheek skin can be a useful source of m-SKP multipotent progenitors. At present lack of sample numbers means that we can only postulate why we were unable to obtain m-SKPs from the lion and red panda cultures. However the giant panda observations point to the value of archiving cells from rare species, and the possibilities for later progenitor cell derivation.

Haematology and Serum Biochemistry Parameters and Variations in the Eurasian Beaver (Castor fiber).

Haematology parameters (N = 24) and serum biochemistry parameters (N = 35) were determined for wild Eurasian beavers (Castor fiber), between 6 months - 12 years old. Of the population tested in this study, N = 18 Eurasian beavers were from Norway and N = 17 originating from Bavaria but now living extensively in a reserve in England. All blood samples were collected from beavers via the ventral tail vein. All beavers were chemically restrained using inhalant isoflurane in 100% oxygen prior to blood sampling. Results were determined for haematological and serum biochemical parameters for the species and were compared between the two different populations with differences in means estimated and significant differences being noted. Standard blood parameters for the Eurasian beaver were determined and their ranges characterised using percentiles. Whilst the majority of blood parameters between the two populations showed no significant variation, haemoglobin, packed cell volume, mean cell haemoglobin and white blood cell counts showed significantly greater values (p<0.01) in the Bavarian origin population than the Norwegian; neutrophil counts, alpha 2 globulins, cholesterol, sodium: potassium ratios and phosphorus levels showed significantly (p<0.05) greater values in Bavarian versus Norwegian; and potassium, bile acids, gamma globulins, urea, creatinine and total calcium values levels showed significantly (p<0.05) greater values in Norwegian versus Bavarian relict populations. No significant differences were noted between male and female beavers or between sexually immature (<3 years old) and sexually mature (≥3 years old) beavers in the animals sampled. With Eurasian beaver reintroduction encouraged by legislation throughout Europe, knowledge of baseline blood values for the species and any variations therein is essential when assessing their health and welfare and the success or failure of any reintroduction program. This is the first study to produce base-line blood values and their variations for the Eurasian beaver.

Echinococcus multilocularis Detection in Live Eurasian Beavers (Castor fiber) Using a Combination of Laparoscopy and Abdominal Ultrasound under Field Conditions.

Echinococcus multilocularis is an important pathogenic zoonotic parasite of health concern, though absent in the United Kingdom. Eurasian beavers (Castor fiber) may act as a rare intermediate host, and so unscreened wild caught individuals may pose a potential risk of introducing this parasite to disease-free countries through translocation programs. There is currently no single definitive ante-mortem diagnostic test in intermediate hosts. An effective non-lethal diagnostic, feasible under field condition would be helpful to minimise parasite establishment risk, where indiscriminate culling is to be avoided. This study screened live beavers (captive, n = 18 or wild-trapped in Scotland, n = 12) and beaver cadavers (wild Scotland, n = 4 or Bavaria, n = 11), for the presence of E. multilocularis. Ultrasonography in combination with minimally invasive surgical examination of the abdomen by laparoscopy was viable under field conditions for real-time evaluation in beavers. Laparoscopy alone does not allow the operator to visualize the parenchyma of organs such as the liver, or inside the lumen of the gastrointestinal tract, hence the advantage of its combination with abdominal ultrasonography. All live beavers and Scottish cadavers were largely unremarkable in their haematology and serum biochemistry with no values suspicious for liver pathology or potentially indicative of E. multilocularis infection. This correlated well with ultrasound, laparoscopy, and immunoblotting, which were unremarkable in these individuals. Two wild Bavarian individuals were suspected E. multilocularis positive at post-mortem, through the presence of hepatic cysts. Sensitivity and specificity of a combination of laparoscopy and abdominal ultrasonography in the detection of parasitic liver cyst lesions was 100% in the subset of cadavers (95%Confidence Intervals 34.24-100%, and 86.7-100% respectively). For abdominal ultrasonography alone sensitivity was only 50% (95%CI 9.5-90.6%), with specificity being 100% (95%CI 79.2-100%). For laparoscopy alone sensitivity was 100% (95% CI 34.2-100%), with specificity also being 100% (95% CI 77.2-100%). Further immunoblotting, PCR and histopathological examination revealed one individual positive for E. multilocularis, whilst the other individual was positive for Taenia martis.