Reduce, Replace, Refine: Determining A Posteriori Reference Intervals for Biochemistry in Hermann's Tortoise (Testudo hermanni).

Biochemical and hematological analyses are important for the assessment of animal health. However, for most wild species their use is hindered by the scarcity of reliable reference intervals. Indeed, collecting body fluids (e.g., blood, urine) in free-ranging animals is often technically challenging. Further, sampling many individuals would be essential to consider major sources of variations, such as species, populations, sex, age, and seasons. One alternative, according to the reduction, refinement, and replacement framework, is to establish reference intervals a posteriori using literature survey and unpublished data. We produced reference intervals for free-ranging Hermann's tortoises (Testudo hermanni), using analyses performed on blood samples collected in previous studies and conservation programs conducted in the field between 2010 and 2016 in southern France (n=195 individuals). Thirteen parameters were analyzed: packed-cell volume, blood concentrations of corticosterone, testosterone, glycemia, cholesterol, triglyceride, urea, uric acid, calcium, sodium, potassium, asparagine aminotransferase (AST), and alanine aminotransferases (ALT). Reference intervals for subgroups defined by sex and season were relevant for corticosterone, triglyceride, and calcium (sex) and cholesterol (season). Comparing our results with those obtained in captive individuals in Germany, except for urea and AST levels the intervals from both free-ranging versus captive tortoises were similar, suggesting that reference intervals established from captive individuals may be suitable for free-ranging populations in this species.

Use of two point-of-care analyzers provides reference intervals of prothrombin time and activated partial prothrombin time in nonanesthetized healthy ferrets (Mustela putorius furo).

OBJECTIVE: Coagulation tests are an essential tool in the diagnosis and management of coagulopathies in mammals. The aim of the current study was to establish reference intervals for prothrombin time (PT) and activated partial PT (aPTT) in healthy ferrets using 2 different point-of-care analyzers (Idexx Coag DX and MS QuickVet Coag Combo). ANIMALS: 86 clinically healthy ferrets under 3 years of age (47 females and 39 males) from 4 breeders and 2 private practices. PROCEDURES: Blood samples were collected from the cranial vena cava in all ferrets without anesthesia and placed in trisodium 3.2% citrated plastic tubes. Sixty-six blood samples from the 4 ferret breeding farms and 1 private practice were analyzed using the Idexx Coag DX and 21 from the other private practice using the MS QuickVet Coag Combo. RESULTS: Reference intervals for the Idexx Coag DX were as follows: aPTT (n = 65), 69.84 to 105.99 seconds; PT (65), 14.44 to 21.98 seconds. Reference intervals for the MS QuickVet Coag Combo were as follows: aPTT (n = 21), 74.90 to 115.50 seconds; PT (21), 18.31 to 23.05 seconds. With both types of analyzers, there was no significant age effect on aPTT and PT. CLINICAL RELEVANCE: This study provided coagulation times for 2 point-of-care analyzers in healthy ferrets as a tool for the diagnosis of coagulopathies.

Prevalence of Reproductive Disorders including Mammary Tumors and Associated Mortality in Female Dogs.

Female dogs, especially intact or neutered lately, are at increased risk for reproductive disorders including mammary tumors (MTs). This retrospective study evaluated the prevalence of reproductive pathology and associated mortality in a cohort of female dogs presented at a single veterinary clinic. The medical records of female dogs born in 2000-2003 were reviewed. The study included 599 cases, of which 293 were followed up until death. Causes of death were analyzed according to the spaying status. Among the 599 female dogs, 306 were intact (51%), 50 (8%) had been spayed before 2 years of age (ES, early spaying), and 243 (41%) after 2 years (LS, late spaying). During their lifetime, 79 dogs (13.2%) developed pyometra, and 160 (26.7%) a mammary tumor. Among the 293 dogs with complete follow-up, 103 (35.1%) had at least one MT during their lifetime, of which 53 (51.5%) died of their mammary cancer. Spayed (ES + LS) female dogs had a 4-fold decreased risk of dying from mammary cancer (OR = 0.23, 95% CI 0.11-0.47, p < 0.0001) compared to intact females. In this low-sterilization rate population, MTs developed in 35.1% of female dogs over their lifetime and was the cause of death in half of them.

Brain Ependymoma in an African Grey Parrot (Psittacus erithacus erithacus).

A 14-year-old unsexed African grey parrot (Psittacus erithacus erithacus) was presented with a 12-hour history of neurological signs and vomiting. The external physical examination of the patient revealed lethargy, moderate hypothermia, a head tilt, and horizontal nystagmus. Whole-body radiographic imaging and blood biochemistry parameters were unremarkable, and a serological test to detect bornavirus was negative. A computed tomography scan of the bird revealed a large cystic lesion located on the fourth ventricle of the brain. In spite of treatment (nonsteroidal anti-inflammatory drugs and antibiotic and antiparasitic therapy), the parrot's health continued to decline, and it was euthanatized 12 days after presentation. A complete postmortem examination was performed on the patient's brain. Histopathological interpretation of tissues submitted described a large neoformation composed of cells arranged in perivascular pseudorosettes. Hyperchromatic nuclei and marked anisokaryosis were suggestive of a malignant tumor. The tissue mass was associated with significant dilation of the fourth ventricle and a severe peripheral gliosis. The histopathological diagnosis of the neoformation was an ependymoma. Ependymomas are glial tumors of the ependymal cells that line the central canal and the ventricles of the brain and are rarely described in mammals. In birds, ependymomas were only described in budgerigars (Melopsittacus undulatus). In human medicine, the recommended treatment is surgical removal of the tumor when possible, followed by radiotherapy.

Ferret Behavior Medicine.

Domestic ferrets (Mustela putorius furo) are common zoologic companion animals and display specific body language and vocalizations. Social interactions, play behavior, and resting periods are important keystone in domestic ferret behavior. Specific housing and environmental enrichment are recommended to preserve the expression of normal behavior and physiology in ferrets. Presence of abnormal behaviors, including aggression, urination and defecation outside the litter box, stereotypies, and absence of play behavior, should be carefully monitored by veterinarians and ferret owners to assess ferret wellness. Specific considerations, such as deafness, poor vision, and hybridation with other mustelids, may play a role in ferret behavior.

Clinical Management of Avian Renal Disease.

Treatment of avian renal disease relies on supportive care, such as fluid therapy and nutritional support. Analgesia and adaptations of the environment are indicated in cases of renal disease associated with painful joints. Other treatments vary with the underlying etiology and may include systemic antibiotics, antifungal therapy, vitamin A supplementation, or chelation therapy.

Multi-host disease management: the why and the how to include wildlife.

In recent years, outbreaks caused by multi-host pathogens (MHP) have posed a serious challenge to public and animal health authorities. The frequent implication of wildlife in such disease systems and a lack of guidelines for mitigating these diseases within wild animal populations partially explain why the outbreaks are particularly challenging. To face these challenges, the French Ministry of Agriculture launched a multi-disciplinary group of experts that set out to discuss the main wildlife specific concepts in the management of MHP disease outbreaks and how to integrate wildlife in the disease management process.This position paper structures the primary specific concepts of wildlife disease management, as identified by the working group. It is designed to lay out these concepts for a wide audience of public and/or animal health officers who are not necessarily familiar with wildlife diseases. The group's discussions generated a possible roadmap for the management of MHP diseases. This roadmap is presented as a cycle for which the main successive step are: step 1-descriptive studies and monitoring; step 2-risk assessment; step 3-management goals; step 4-management actions and step 5-assessment of the management plan. In order to help choose the most adapted management actions for all involved epidemiological units, we integrated a decision-making framework (presented as a spreadsheet). This tool and the corresponding guidelines for disease management are designed to be used by public and health authorities when facing MHP disease outbreaks. These proposals are meant as an initial step towards a harmonized transboundary outbreak response framework that integrates current scientific understanding adapted to practical intervention.

Accuracy and precision of measurements performed on three-dimensional printed pelvises when compared to computed tomography measurements.

The preoperative contouring of plates decreases the duration of surgery and improves the quality of the reduction of pelvic fractures. Patient-tailored three-dimensionally printed pelvises might be an interesting tool for achieving that purpose. Currently, no study has evaluated the accuracy of measurements performed on three-dimensional printed models in comparison with computed tomography data for complex bones, such as the pelvis. This study examined whether the measurements obtained on pelvises printed using dual-material fused deposition modeling technology are not significantly different from those obtained on computed tomography images. The computed tomography images of the pelvic region from 10 dogs were used to produce three-dimensionally printed models with a dual-material fused deposition-modeling process. Four segments were measured on both three-dimensionally printed models and computed tomography images. The measurements were performed by three observers and repeated twice. Concordance correlation coefficients were used to assess the precision and accuracy of the measurements as well as evaluate the agreement between the methods. The accuracy of measurements between the methods was > 0.99 for all measurements. The precision was almost perfect for AE (0.996), substantial for BD and BC (0.963 and 0.958, respectively), and moderate for CD (0.912). These results indicate that, despite some minor variations, the measurements performed on printed models reproduced the computed tomography data reliably.

Agreement between Repeated Subjective Visual Evaluations of Body Condition of St. Lawrence Beluga Whale ( Delphinapterus leucas) Carcasses Using a Visual Analog Scale.

The determination of body condition is a fundamental component of an evaluation during postmortem examinations of cetaceans. Three veterinarians experienced in beluga postmortem techniques subjectively evaluated 62 carcasses of beluga whales ( Delphinapterus leucas) in duplicate, using a visual analog scale. The result of evaluation and scoring was repeatable in cachectic animals and animals in very good condition but did not capture the continuum of body condition determinations between the obvious visual conditions.

Evidence-Based Advances in Aquatic Animal Medicine.

Fish and aquatic invertebrates deserve evidence-based medicine. Pharmacologic information is available; most pharmacokinetic studies are derived from the aquaculture industry and extrapolated to ornamental fish. Conversely, advanced diagnostics and information regarding diseases affecting only ornamental fish and invertebrates require more peer-reviewed experimental studies; the examples of carp edema virus, sea star wasting disease, seahorse nutrition, and gas bubble disease of fish under human care are discussed. Antinociception is also a controversial topic of growing interest in aquatic animal medicine. This article summarizes information regarding new topics of interest in companion fish and invertebrates and highlights some future avenues for research.

Evaluation of High Dosages of Oral Meloxicam in American Kestrels ( Falco sparverius ).

To evaluate the toxicity of short-term high doses of meloxicam in American kestrels ( Falco sparverius ), 32 male captive-born, 1- to 4-year-old American kestrels were randomly assigned to 4 groups: 3 groups treated with meloxicam (n = 9 per group) and a control group (n = 5). Meloxicam was administered orally via feeding tube in the proventriculus at 2, 10, and 20 mg/kg every 12 hours for 7 days for the treatment groups, while the control group received saline solution. The birds were evaluated for the presence of clinical signs, abnormalities in the complete blood cell count and in the plasma biochemical panel for the 20-mg/kg group, and gross and histopathologic lesions. No clinical signs or mortality were observed in any group. No significant differences of clinical relevance were found in results of the packed cell volume, total solids, and biochemical panel, and no evidence of renal toxicity was found in the treatment or control groups. A significant correlation was found between hepatic lipidosis and meloxicam dose (P = .02). Two of 9 birds in the 20-mg/kg group developed gastric ulcers, although this result was not significant. None of the birds in the 2- and 10-mg/kg groups had similar lesions. Finally, meloxicam dosages up to 20 mg/kg did not result in nephrotoxicity in American kestrels. Further toxicologic studies to evaluate hepatotoxicity and gastrotoxicity of meloxicam in avian species are needed.

Microsporidiosis in Vertebrate Companion Exotic Animals.

Veterinarians caring for companion animals may encounter microsporidia in various host species, and diagnosis and treatment of these fungal organisms can be particularly challenging. Fourteen microsporidial species have been reported to infect humans and some of them are zoonotic; however, to date, direct zoonotic transmission is difficult to document versus transit through the digestive tract. In this context, summarizing information available about microsporidiosis of companion exotic animals is relevant due to the proximity of these animals to their owners. Diagnostic modalities and therapeutic challenges are reviewed by taxa. Further studies are needed to better assess risks associated with animal microsporidia for immunosuppressed owners and to improve detection and treatment of infected companion animals.

Comparison of a human portable glucometer and an automated chemistry analyzer for measurement of blood glucose concentration in pet ferrets (Mustela putorius furo).

This study compared blood glucose concentrations measured with a portable blood glucometer and a validated laboratory analyzer in venous blood samples of 20 pet ferrets (Mustela putorius furo). Correlation and agreement were evaluated with a Bland-Altman plot method and Lin's concordance correlation coefficient. Blood glucose concentrations measured with the laboratory analyzer and the glucometer ranged from 1.9 to 8.6 mmol/L and from 0.9 to 9.2 mmol/L, respectively. The glucometer had a poor agreement and correlation with the laboratory analyzer (bias, -0.13 mmol/L; level of agreement, -2.0 to 3.6 mmol/L, concordance correlation coefficient 0.665). The relative sensitivity and specificity of the portable blood glucometer for detection of hypoglycemia were 100% (95% CI: 66% to 100%) and 50% (95% CI: 20% to 80%), respectively. Positive and negative predictive values were 67% (95% CI: 39% to 87%) and 100% (95% CI: 46% to 100%), respectively. Based on these results, clinicians are advised to be cautious when considering the results from this handheld glucometer in pet ferrets, and blood glucose concentrations should be determined with a laboratory analyzer validated for this species.

Comparaison entre un lecteur de glycémie portable pour humain et un analyseur automatisé de biochimie dans le but d'évaluer de la glycémie chez des furets domestiques(Mustela putorius furo). L'objectif de l'étude était de comparer les valeurs de glycémie mesurées par un glycomètre portable et un analyseur de laboratoire certifié pour des prélèvements sanguins veineux effectués sur 20 furets de compagnie (Mustela putorius furo). L'équivalence des méthodes a été évaluée grâce à un diagramme de Bland-Altman et au coefficient de corrélation de concordance de Lin. Les glycémies mesurées par l'analyseur de laboratoire et le glycomètre étaient respectivement comprises entre 1,9 à 8,6 mmol/L et de 0,9 à 9,2 mmol/L. Les degrés d'agrément et de corrélation entre le glycomètre et l'analyseur de laboratoire étaient faibles (biais, −0,13 mmol/L; niveau d'agrément, −2,0 à 3,6 mmol/L, coefficient de corrélation de concordance 0,665). La sensibilité et la spécificité du glycomètre concernant la détection d'hypoglycémie étaient respectivement de 100 % (95 % CI : 66­100 %) et de 50 % (95 % CI : 20­80 %) et les valeurs predictives positive et negative étaient respectivement de 67 % (95 % CI : 39­87 %) et de 100 % (95 % CI : 46­100 %). En s'appuyant sur ces résultats, l'utilisation d'un glycomètre portable devraient être réalisée avec précaution en pratique chez les furets de compagnie et les valeurs de glycémie devraient être déterminées par un analyseur de laboratoire certifié pour cette espèce.(Traduit par les auteurs).

Tetralogy of Fallot in a 6-year-old albino ferret (Mustela putorius furo).

Tetralogy of Fallot associated with bidirectional shunting across a large ventricular septal defect, was found in a 6-year-old ferret. The prognosis associated with tetralogy of Fallot is usually poor. This case is interesting given the advanced age of the ferret. The bidirectional shunting, responsible for an acyanotic disease, may explain the unexpected prolonged survival in this ferret.

Tétralogie de Fallot chez un furet de 6 ans(Mustela putorius furo) . Une tétralogie de Fallot, associée à un shunt bidirectionnel au travers d'une communication interventriculaire béante, a été découverte chez un furet de six ans. Le pronostic de la tétralogie de Fallot est habituellement sombre. La survie prolongée de l'animal est intéressante. Il est probable que le shunt bidirectionnel, responsable d'une maladie acyanotique, explique l'espérance de vie prolongée de ce furet.(Traduit par les auteurs).

Relationship between red vent syndrome and anisakid larvae burden in wild atlantic salmon (salmo salar).

The pathogenesis of the recently recognized "red vent syndrome" in wild Atlantic salmon (Salmo salar) is not fully understood. Pathologic observations indicate that this syndrome is associated with the presence of nonencapsulated larvae of the nematode Anisakis simplex in the body wall, the lower intestinal wall, and the visceral cavity surrounding the vent region. We evaluated the relationship between the occurrence of red vent syndrome and intensity of infection with Anisakis sp. larvae in naturally infected fish. Salmon caught by sport anglers were opportunistically evaluated to detect red vent syndrome. We included 106 salmon with red vent syndrome and 98 without red vent syndrome in this study. Intensity of infection was established by counting the total number of perivisceral larvae and by determining the number of larvae per gram in 10 g of pepsin-digested perianal tissue. The severity of inflammatory changes was also evaluated in standard histologic sections of the perianal area using a semiquantitative scale. Salmon with red vent syndrome had significantly higher intensity of inflammation than salmon without red vent syndrome (P=0.008). The odds of having red vent syndrome increased with the number of perianal larvae per gram of perianal tissue (P=0.002; odds ratio [OR]=1.12; 95% confidence interval: [1.05; 1.22]) but not with the number of perivisceral larvae, fish length, or gender. Although these results support the association between this syndrome and intensity of infection by A. simplex, the relationship is not strong (OR near 1), suggesting that the clinical expression of red vent syndrome at an individual level, and the emergence of this disease on a global scale, must be determined by other factors, such as timing of infection.

Oral rabies vaccination of raccoons and striped skunks with ONRAB® baits: multiple factors influence field immunogenicity.

Multiple control methods have been used in North America to manage the spread of rabies caused by the raccoon (Procyon lotor) rabies virus variant (RRVV). Recently, oral vaccination with ONRAB(®) vaccine baits, which contain an adenovirus rabies glycoprotein recombinant, has been made available as an additional tool for rabies control. Our objectives were to estimate rabies antibody prevalence in wild-caught raccoons and striped skunks (Mephitis mephitis), and identify factors influencing the probability of being antibody positive at the individual level in these species, following oral rabies vaccination (ORV) campaigns in which ONRAB was distributed aerially in 2007-2009 in southern Québec, Canada. Following the aerial distribution of 43-155 ONRAB baits/km(2), the annual percentages of antibody-positive raccoons and skunks varied between 35% and 56% and 11% and 17%, respectively. In raccoons, the probability of being antibody positive was positively associated with age and density of ONRAB distributed, and influenced by the number of previous ORV campaigns conducted. Conversely, this probability was negatively associated with estimated abundance of raccoons in the trapping cell and proportion of residential areas near the raccoon capture location. None of the variables examined explained variation in the probability of being antibody positive in skunks. Our results indicate that the ONRAB density applied during ORV campaigns should be adjusted to account for variations in raccoon population density and presence of residential areas to increase the likelihood of creating an effective immunological barrier against RRVV. The high percentage of juvenile raccoons (annual mean =45 ± 3 [SE]%) and skunks (66 ± 2%) captured during post-ORV monitoring suggests that ORV campaigns should be conducted at least annually to account for the recruitment of naïve individuals into the populations. In Québec, the increasing use of ONRAB coincided with the elimination of rabies caused by RRVV. Nonetheless, our results indicate that improvements to this vaccine bait and/or the distribution techniques are required to increase its efficacy, especially in striped skunks.

From science to action and from action to science: the Nunavik Trichinellosis Prevention Program.

OBJECTIVES: During the 1980s, walrus-meat consumption caused infections with the parasite Trichinella nativa in Nunavik inhabitants. In response to these events, stakeholders set up the community-based Nunavik Trichinellosis Prevention Program (NTPP). The objectives of the present communication are to review the NTPP, describe how science and action were interwoven in its development and identify its assets and limitations. Study design. Descriptive study. METHODS: The NTPP relies on a pooled digestion assay of tongue samples taken from each harvested walrus. The public health recommendations depend on the results of the analyses: infected walrus meat should be destroyed; parasite-free meat may be eaten raw or cooked. RESULTS: All communities involved in the walrus hunt participate in the NTPP and a high percentage of harvested walruses are included in the NTPP. Infected animals account for 2.9% of the walruses tested (20/694) since 1992. The NTPP permitted the early management of a trichinellosis event in 1997. Since then, it prevented the new occurrence of outbreaks related to walruses hunted by Nunavimmiut. CONCLUSIONS: The absence of recent major outbreaks of trichinellosis in Nunavik may reasonably be attributed to the NTPP. The success of the program stands on many facilitating factors such as the nature of the disease and its source, the existence of an efficient analytic method, the strong involvement of the different partners including direct resource users, as well as the comprehensive bidirectional science-to-action approach that has been followed.

Emaciation and larval filarioid nematode infection in boreal owls (Aegolius funereus).

Microfilariae are considered non-pathogenic in wild birds. The objective of the current communication is to report host reactions to microfilarial infection of unusual intensity in emaciated boreal owls (Aegolius funereus). An unusually large number of boreal owls (n = 21) were submitted to the Canadian Cooperative Wildlife Health Center-Quebec Region for post-mortem examination during the winter of 2009. Nineteen out of 21 birds were considered emaciated based on atrophy of adipose tissue and pectoral muscles and suboptimal weight. A microscopic examination of a subset of nine owls revealed the presence of microfilariae in six owls. Three of the birds with a heavy parasite burden had masses of larval nematodes obstructing large vessels of the lungs. The emaciated owls are believed to have died from starvation due to a cyclic decrease in prey abundance in the boreal forest. This cycle also drives winter movements of boreal owls to urbanized areas of southern Quebec, presumably accounting for the large number of birds submitted in 2009. In the most severely infected owls, the extreme microfilarial burden might have caused an alteration in circulatory dynamics, gaseous exchanges and also probably some metabolic cost. Consequently, microfilariae could have significantly contributed to the death of some of these owls.

Low sensitivity of antemortem gill biopsies for the detection of subclinical Pseudodactylogyrus bini infestations in American eels (Anguilla rostrata).

The objectives of this study were twofold: to determine the sensitivity of gill biopsies used for the screening of Pseudodactylogyrus bini in subclinically infested American eels (Anguilla rostrata) and to evaluate the effect of bath anesthesia on the proportion of infested eels. Fifty-four eels were euthanized following either intracoelomic injection of T61 or deep anesthesia with tricaine methanesulfonate. Proportions of eels infested by P. bini diagnosed by gill biopsies taken through the gill slit and by microscopic examination of the entire gill arches were compared. The euthanasia protocol had no significant effect on parasite abundance. The prevalence of monogeneans detected by examination of nonterminal antemortem gill biopsies and by complete gill examinations was 20% and 70%, respectively. The sensitivity of gill biopsies (29%) to detect P. bini in subclinically infested eels is therefore low, and should be used for group screening rather than individual screening.