Amphibian Dermatology.

Amphibians are susceptible to a multitude of skin disorders, many of which can appear grossly similar. The most common clinical presentations include hyperemia, discoloration, dermal mass, ulceration, and necrosis. Many amphibian skin diseases are related to captive husbandry. The diagnostic process starts with environmental evaluations, a full history, physical examination and sampling for direct observation, histology, polymerase chain reaction testing, and bacterial and fungal culture. This review emphasizes the main conditions encountered in amphibian dermatology.

CardiOvascular examination in awake Orangutans (Pongo pygmaeus pygmaeus): Low-stress Echocardiography including Speckle Tracking imaging (the COOLEST method).

INTRODUCTION: Cardiovascular diseases have been identified as a major cause of mortality and morbidity in Borneo orangutans (Pongo pygmaeus pygmaeus). Transthoracic echocardiography is usually performed under anesthesia in great apes, which may be stressful and increase risks of peri-anesthetic complications in case of cardiac alteration. The aim of the present pilot study was hence to develop a quick and non-stressful echocardiographic method (i.e., the COOLEST method) in awake Borneo orangutans (CardiOvascular examination in awake Orangutans: Low-stress Echocardiography including Speckle Tracking imaging) and assess the variability of corresponding variables. MATERIALS AND METHODS: Four adult Borneo orangutans trained to present their chest to the trainers were involved. A total of 96 TTE examinations were performed on 4 different days by a trained observer examining each orangutan 6 times per day. Each examination included four two-dimensional views, with offline assessment of 28 variables (i.e., two-dimensional (n = 12), M-mode and anatomic M-mode (n = 6), Doppler (n = 7), and speckle tracking imaging (n = 3)), representing a total of 2,688 measurements. A general linear model was used to determine the within-day and between-day coefficients of variation. RESULTS: Mean±SD (minimum-maximum) images acquisition duration was 3.8±1.6 minutes (1.3-6.3). All within-day and between-day coefficients of variation but one (n = 55/56, 98%) were <15%, and most (51/56, 91%) were <10% including those of speckle tracking systolic strain variables (2.7% to 5.4%). DISCUSSION: Heart morphology as well as global and regional myocardial function can be assessed in awake orangutans with good to excellent repeatability and reproducibility. CONCLUSIONS: This non-stressful method may be used for longitudinal cardiac follow-up in awake orangutans.

Reproductive Techniques for Ovarian Monitoring and Control in Amphibians.

Ovarian control and monitoring in amphibians require a multi-faceted approach. There are several applications that can successfully induce reproductive behaviors and the acquisition of gametes and embryos for physiological or molecular research. Amphibians contribute to one-quarter to one-third of vertebrate research, and of interest in this context is their contribution to the scientific community's knowledge of reproductive processes and embryological development. However, most of this knowledge is derived from a small number of species. In recent times, the decimation of amphibians across the globe has required increasing intervention by conservationists. The captive recovery and assurance colonies that continue to emerge in response to the extinction risk make existing research and clinical applications invaluable to the survival and reproduction of amphibians held under human care. The success of any captive population is founded on its health and reproduction and the ability to develop viable offspring that carry forward the most diverse genetic representation of their species. For researchers and veterinarians, the ability to monitor and control ovarian development and health is, therefore, imperative. The focus of this article is to highlight the different assisted reproductive techniques that can be used to monitor and, where appropriate or necessary, control ovarian function in amphibians. Ideally, any reproductive and health issues should be reduced through proper captive husbandry, but, as with any animal, issues of health and reproductive pathologies are inevitable. Non-invasive techniques include behavioral assessments, visual inspection and palpation and morphometric measurements for the calculation of body condition indices and ultrasound. Invasive techniques include hormonal injections, blood sampling, and surgery. Ovarian control can be exercised in a number of ways depending on the application required and species of interest.

Persistent fibrosis, hypertrophy and sarcomere disorganisation after endoscopy-guided heart resection in adult Xenopus.

Models of cardiac repair are needed to understand mechanisms underlying failure to regenerate in human cardiac tissue. Such studies are currently dominated by the use of zebrafish and mice. Remarkably, it is between these two evolutionary separated species that the adult cardiac regenerative capacity is thought to be lost, but causes of this difference remain largely unknown. Amphibians, evolutionary positioned between these two models, are of particular interest to help fill this lack of knowledge. We thus developed an endoscopy-based resection method to explore the consequences of cardiac injury in adult Xenopus laevis. This method allowed in situ live heart observation, standardised tissue amputation size and reproducibility. During the first week following amputation, gene expression of cell proliferation markers remained unchanged, whereas those relating to sarcomere organisation decreased and markers of inflammation, fibrosis and hypertrophy increased. One-month post-amputation, fibrosis and hypertrophy were evident at the injury site, persisting through 11 months. Moreover, cardiomyocyte sarcomere organisation deteriorated early following amputation, and was not completely recovered as far as 11 months later. We conclude that the adult Xenopus heart is unable to regenerate, displaying cellular and molecular marks of scarring. Our work suggests that, contrary to urodeles and teleosts, with the exception of medaka, adult anurans share a cardiac injury outcome similar to adult mammals. This observation is at odds with current hypotheses that link loss of cardiac regenerative capacity with acquisition of homeothermy.

Reproductive Medicine in Amphibians.

Reproduction of amphibians includes ovulation, spermiation, fertilization, oviposition, larval stage and development, and metamorphosis. A problem at any stage could lead to reproductive failure. To stimulate reproduction, environmental conditions must be arranged to simulate changes in natural habits. Reproductive life history is well documented in amphibians; a thorough knowledge of this subject will aid the practitioner in diagnosis and treatment. Technologies for artificial reproduction are developing rapidly, and some protocols may be transferable to privately kept or endangered species. Reproductive tract disorders are rarely described; no bacterial or viral diseases are known that specifically target the amphibian reproductive system.

PROPOSED SIMPLE METHOD FOR ELECTROCARDIOGRAM RECORDING IN FREE-RANGING ASIAN ELEPHANTS (ELEPHAS MAXIMUS).

Electrocardiography represents a relevant diagnostic tool for detecting cardiac disease in animals. Elephants can present various congenital and acquired cardiovascular diseases. However, few electrophysiologic studies have been reported in captive elephants, mainly due to challenging technical difficulties in obtaining good-quality electrocardiogram (ECG) tracings, and no data are currently available for free-ranging Asian elephants (Elephas maximus). The purpose of this pilot prospective study was to evaluate the feasibility of using a simple method for recording ECG tracings in wild, apparently healthy, unsedated Asian elephants (n = 7) in the standing position. Successful six-lead recordings (I, II, III, aVR, aVL, and aVF) were obtained, with the aVL lead providing the best-quality tracings in most animals. Variables measured in the aVL lead included heart rate, amplitudes and duration of the P waves, QRS complexes, T and U waves, and duration of the PR, QT, and QU intervals. A negative deflection following positive P waves, representative of an atrial repolarization wave (Ta wave), was observed for five out of the seven elephants.

Surgery in Amphibians.

Amphibian surgery has been especially described in research. Since the last decade, interest for captive amphibians has increased, so have the indications for surgical intervention. Clinicians should not hesitate to advocate such manipulations. Amphibian surgeries have no overwhelming obstacles. These patients heal well and tolerate blood loss more than higher vertebrates. Most procedures described in reptiles (mostly lizards) can be undertaken in most amphibians if equipment can be matched to the patients' size. In general, the most difficult aspect would be the provision of adequate anesthesia.

PREVALENCE OF VALVULAR REGURGITATIONS IN CLINICALLY HEALTHY CAPTIVE LEOPARDS AND CHEETAHS: A PROSPECTIVE STUDY FROM THE WILDLIFE CARDIOLOGY (WLC) GROUP (2008-2013).

The purpose of this prospective study was to evaluate transthoracic echocardiograms from clinically healthy large felids for the presence of valvular regurgitations (VR). Physiologic VR commonly occur in normal dogs and cats, but the percentage of large felids with VR has not been previously reported. During a 5-yr study period (2008-2013), 28 healthy animals were evaluated under general anesthesia: 16 cheetahs (Acinonyx jubatus soemmeringuii) with a mean age of 1.5±0.8 yr (range 0.7-3.5 yr), 5 Amur leopards (Panthera pardus orientalis), 1 snow leopard (Uncia uncia), and 6 clouded leopards (Neofelis nebulosa). For this study, all the leopards were gathered in one so-called "leopards group" with a mean age of 2.8±3.4 yr (range 0.3-10.7 yr). All valves observed in each view were examined for evidence of regurgitant jets and turbulent blood flow using the color-flow Doppler mode. Valves were also examined for structural changes. Mitral valve and aortic cusp abnormalities were considered to be of congenital origin. Mitral valve lesions led to mitral insufficiency in all the felids. Aortic cusp abnormalities led to aortic regurgitation in 94% of the cheetahs and 67% of the leopards. Leopards showed a predominance of early systolic mitral regurgitations, whereas all the mitral regurgitation jets in cheetahs were holosystolic. Tricuspid regurgitation was found in 81% of the cheetahs and in 50% of the leopards, whereas pulmonic regurgitation was detected in 44% of the cheetahs and 33% of the leopards. Interestingly, none of these tricuspid and pulmonic regurgitations were associated with two-dimensional structural valve abnormalities, thus suggesting their physiologic origin, as described in humans, cats, and dogs. In conclusion, subclinical valvular diseases are common in apparently healthy leopards and cheetahs. Longitudinal follow-up of affected animals is therefore required to assess their clinical outcome.

Endoscopy and Endosurgery in Nonhuman Primates.

Endoscopy in nonhuman primates (NHPs) has resulted in improvements in research and clinical care for more than 4 decades. The indications and procedures are the same as in humans and the approach is similar to that in dogs, cats, and humans. Selected procedures are discussed including rhinoscopy, tracheobronchoscopy, upper gastrointestinal endoscopy, laparoscopy, and endoscopic salpingectomy. This short overview provides practitioners with pragmatic elements for safe and effective endoscopy in NHPs.

Endoscopy in Amphibians.

Despite advances in exotic animal endoscopy, descriptions involving amphibians are scarce. Amphibian endoscopy shares some similarities with reptiles, especially in lizards. Selected procedures are discussed, including stomatoscopy, gastroscopy, coelioscopy, and biopsy of coelomic organs and lesions. This short overview provides the practitioner with pragmatic advice on how to conduct safe and effective endoscopic examinations in amphibians.

Propagation method of saving valuable strains from a Mycobacterium liflandii infection in Western clawed frogs (Silurana tropicalis).

Mycobacterium liflandii has been responsible for an emerging infection reported in the international trade of Western clawed frogs (Silurana tropicalis). This study shows that this mycolactone-producing Mycobacterium (MPM) has expanded its distribution range to France. The results of this study suggest that the use of in vitro fertilization to maintain genetic lines could be a temporary solution for valuable S. tropicalis propagation.

Atherosclerosis associated with pericardial effusion in a central bearded dragon (Pogona vitticeps).

Atherosclerosis is a common disease in pet birds, particularly in psittacines, and is frequently found when performing postmortem examinations on adult and old dogs, in which it is mainly associated with endocrine diseases, such as hypothyroidism and diabetes mellitus. However, atherosclerosis is poorly documented in reptiles and consequently poorly understood. In the current case report, atherosclerosis and pericardial effusion were diagnosed in a 2-year-old male central bearded dragon (Pogona vitticeps) based on ultrasound visualization, necropsy, and histologic examination.

Successful treatment of a congenital pulmonic valvular stenosis in a snow leopard (Uncia uncia) by percutaneous balloon valvuloplasty.

A 3-yr-old intact female snow leopard (Uncia uncia) was evaluated for progressive apathy, lethargy, and decreased appetite. Cardiac auscultation revealed a left basal grade IV/VI systolic ejection murmur, and an echocardiogram confirmed a severe pulmonic valvular stenosis (pressure gradient of 98 mm Hg). The lesion was managed by balloon valvuloplasty, resulting in a marked pressure gradient reduction (30 mm Hg). The cat recovered well, and clinical signs resolved. This is the first description of a pulmonary valve stenosis and management with balloon valvuloplasty in a wild felid.

Treatment of a retroperitoneal abscess by omentalization in an orangutan (Pongo pygmaeus pygmaeus).

A 38-yr-old orangutan (Pongo pygmaeus pygmaeus) presented with chronic lethargy and difficulty in locomotion that progressed to weakness, anorexia, and permanent dorsal and/or lateral recumbency. The orangutan was immobilized with ketamine. Abdominal ultrasonography revealed a mass in the caudal portion of the abdomen. Exploratory surgery was performed, but the mass could not be resected. Instead, the mass was drained and omentalized in an attempt to establish continuous drainage after surgery. The only complication was a wound infection that was treated locally with a disinfectant and installation of a drain that was changed every 2 days under anesthesia. Omentalization was successful in providing continuous fluid drainage for this retroperitoneal abscess and required minimal postoperative handling of the animal.

Chromosome evolution in the subtribe Bovina (Mammalia, Bovidae): the karyotype of the Cambodian banteng (Bos javanicus birmanicus) suggests that Robertsonian translocations are related to interspecific hybridization.

Three subspecies of banteng (Bos javanicus) have been described: B. j. javanicus in Java, B. j. lowi in Borneo, and B. j. birmanicus in Cambodia, Lao PDR, Myanmar, Thailand and Vietnam. In this paper we provide the first description of the karyotype of the Cambodian banteng. The chromosomal complement of B. j. birmanicus differs from that of B. j. javanicus, which was previously found to be similar to that of cattle, Bos taurus (2n = 60). The Cambodian banteng karyotype has a diploid number of 2n = 56 (FN = 62) and the karyotype consists of 26 pairs of acrocentric chromosomes and two pairs of submetacentric chromosomes. Comparisons with other species of the subtribe Bovina show that the two pairs of bi-armed chromosomes resulted from two centric fusions involving the equivalent of cattle chromosomes 1 and 29, and 2 and 28, respectively. Cross-species fluorescence in-situ hybridization (FISH) with B. taurus whole chromosome paints and satellite DNA I probes was used to identify the chromosomes involved in the translocations, and their orientation. We suggest that Robertsonian translocations (1;29) and (2;28) have been fixed in the common ancestor of Cambodian banteng as a consequence of hybridization with the kouprey (Bos sauveli) during the Pleistocene epoch.

Transgenesis procedures in Xenopus.

Stable integration of foreign DNA into the frog genome has been the purpose of several studies aimed at generating transgenic animals or producing mutations of endogenous genes. Inserting DNA into a host genome can be achieved in a number of ways. In Xenopus, different strategies have been developed which exhibit specific molecular and technical features. Although several of these technologies were also applied in various model organizms, the attributes of each method have rarely been experimentally compared. Investigators are thus confronted with a difficult choice to discriminate which method would be best suited for their applications. To gain better understanding, a transgenesis workshop was organized by the X-omics consortium. Three procedures were assessed side-by-side, and the results obtained are used to illustrate this review. In addition, a number of reagents and tools have been set up for the purpose of gene expression and functional gene analyses. This not only improves the status of Xenopus as a powerful model for developmental studies, but also renders it suitable for sophisticated genetic approaches. Twenty years after the first reported transgenic Xenopus, we review the state of the art of transgenic research, focusing on the new perspectives in performing genetic studies in this species.

Mycobacterium szulgai infection in a captive population of African clawed frogs (Xenopus tropicalis).

A colony of captive Xenopus tropicalis became infected with Mycobacterium szulgai. Clinical signs, when observed, were lethargy, weight loss, and emaciation. Visceral granulomas were common findings at laparoscopy and necropsy. The diagnosis of mycobacteriosis was based on histologic appearance and Ziehl-Neelsen staining of tissues. The identification of M. szulgai organisms was based on comparison of the 16S rRNA gene sequence with several GenBank databases. There have been no reports of this mycobacterial species as the causative agent of naturally occurring disease in amphibians.

Ebstein anomaly in a meerkat (Suricata suricatta).

An 8-yr-old male meerkat (Suricata suricatta) presented with sudden lethargy. A globular-shaped heart associated with pleural effusion on chest thoracic radiographs was consistent with congestive heart failure, and echocardiography was performed under general anesthesia. It revealed an Ebstein anomaly, with mild pericardial effusion and marked right heart enlargement. The animal was treated with imidapril chlorydrate. After 4 mo of treatment, thoracic radiographs still showed right-sided cardiomegaly; however, the animal appeared clinically normal.