Jack Kottwitz

Enhancing captive Indian rhinoceros genetics via artificial insemination of cryopreserved sperm

The objective of this study was to design an artificial insemination (AI) protocol using cryopreserved spermatozoa to obtain pregnancies in captive Indian rhinoceroses (Rhinoceros unicornis). Four methods developed varied by timing and approach, as follows; Method 1: females (n=2) were inseminated pre- and post-ovulation under general anesthesia, Method 2: females (n=2) were inseminated pre-ovulation without anesthetic via endoscopy, Method 3: females (n=1) were inseminated pre-ovulation without anesthetic via manual insertion of an insemination catheter, Method 4: females (n=2) were inseminated same as Method 3 with the addition of standing sedation. Semen deposition site varied as a result of changes in AI technology and experience. All females conceived following intrauterine AI using three methods. Four pregnancies (n=3 females) produced via Method 3 and 4 resulted in term births (n=2 male calves, n=2 female calves) at 481.8±12.8days post-AI. Unfortunately, two early pregnancy losses were documented in a fourth female conceiving via Method 2. Pregnancy rates were 0%, 22%, 17%, and 50% for Method 1-4, respectively. Method 3 and 4 rates improved to 29% and 67%, respectively when accounting for AIs conducted only on ovulatory estrous cycles. Spermatozoa (n=5 males) were cryopreserved 0.3-9.3 y prior to successful AI procedures. The lowest dose of frozen-thawed sperm resulting in conception was 500×10(6) motile sperm. Mean time from AI to ovulation in conceptive and non-conceptive cycles was 26±11.8h and 66±80.7h, respectively.

Formalizing the definition of meta‐analysis in Molecular Ecology

Meta‐analysis, the statistical synthesis of pertinent literature to develop evidence‐based conclusions, is relatively new to the field of molecular ecology, with the first meta‐analysis published in the journal Molecular Ecology in 2003 (Slate & Phua 2003). The goal of this article is to formalize the definition of meta‐analysis for the authors, editors, reviewers and readers of Molecular Ecology by completing a review of the meta‐analyses previously published in this journal. We also provide a brief overview of the many components required for meta‐analysis with a more specific discussion of the issues related to the field of molecular ecology, including the use and statistical considerations of Wrights FST and its related analogues as effect sizes in meta‐analysis. We performed a literature review to identify articles published as ‘meta‐analyses’ in Molecular Ecology, which were then evaluated by at least two reviewers. We specifically targeted Molecular Ecology publications because as a flagship journal in this field, meta‐analyses published in Molecular Ecology have the potential to set the standard for meta‐analyses in other journals. We found that while many of these reviewed articles were strong meta‐analyses, others failed to follow standard meta‐analytical techniques. One of these unsatisfactory meta‐analyses was in fact a secondary analysis. Other studies attempted meta‐analyses but lacked the fundamental statistics that are considered necessary for an effective and powerful meta‐analysis. By drawing attention to the inconsistency of studies labelled as meta‐analyses, we emphasize the importance of understanding the components of traditional meta‐analyses to fully embrace the strengths of quantitative data synthesis in the field of molecular ecology.

NONBACTERIAL THROMBOTIC ENDOCARDITIS IN A FERRET (MUSTELA PUTORIUS FURO)

Abstract A 4-yr-old male ferret (Mustela putorius furo) was presented with a 3-day history of pelvic limb ataxia and weakness. A cellulitis associated with a deep bite wound was identified and initially treated with antibiotics and anti-inflammatory medication. Two weeks later, a grade IV/VI cardiac murmur was identified on physical examination. Echocardiographic examination revealed irregular thickening of the aortic valve leaflets, with normal chamber dimensions and normal systolic function. The ferrets physical condition rapidly deteriorated and it was subsequently euthanatized. Myxomatous degeneration of the aortic valve leaflets, with ulceration and vegetative lesions, and multiorgan infarctions were identified during necropsy. A bacterial etiologic agent was not identified from blood culture or histopathology. Lesions present were consistent with nonbacterial thrombotic endocarditis, a novel condition in the ferret.

Neocortical neuronal morphology in the newborn giraffe (Giraffa camelopardalis tippelskirchi) and African elephant (Loxodonta africana)

Although neocortical neuronal morphology has been documented in the adult giraffe (Giraffa camelopardalis tippelskirchi) and African elephant (Loxodonta africana), no research has explored the cortical architecture in newborns of these species. To this end, the current study examined the morphology of neurons from several cortical areas in the newborn giraffe and elephant. After cortical neurons were stained with a modified Golgi technique (N = 153), dendritic branching and spine distributions were analyzed by using computer‐assisted morphometry. The results showed that newborn elephant neurons were considerably larger in terms of all dendritic and spine measures than newborn giraffe neurons. Qualitatively, neurons in the newborns appeared morphologically comparable to those in their adult counterparts. Neurons in the newborn elephant differed considerably from those observed in other placental mammals, including the giraffe, particularly with regard to the morphology of spiny projection neurons. Projection neurons were observed in both species, with a much larger variety in the elephant (e.g., flattened pyramidal, nonpyramidal multipolar, and inverted pyramidal neurons). Although local circuit neurons (i.e., interneurons, neurogliaform, Cajal–Retzius neurons) resembled those observed in other eutherian mammals, these were usually spiny, which contrasts with their adult, aspiny equivalents. Newborn projection neurons were smaller than the adult equivalents in both species, but newborn interneurons were approximately the same size as their adult counterparts. Cortical neuromorphology in the newborn giraffe is thus generally consistent with what has been observed in other cetartiodactyls, whereas newborn and adult elephant morphology appears to deviate substantially from what is commonly observed in other placental mammals. J. Comp. Neurol. 524:257–287, 2016.

Meta-Analysis of the Effects of Adhesion Barriers on Adhesion Formation in the Horse.

OBJECTIVE To determine the efficacy of adhesion barriers in horses using quantitative statistical analysis. STUDY DESIGN Meta-analytical review. METHODS A search using PubMed/MEDLINE and Google Scholar was performed, followed by secondary searches of veterinary trade journals, bibliographies of relevant articles, manufacturer websites, and technical reference guides. Randomized experimental trials in healthy horses were considered that included both a treatment and control group. The endpoint required was euthanasia or laparoscopy to identify adhesion formation. A meta-analysis was performed using a random effects model, with the effect size calculated as an odds ratio (OR) with 95% confidence intervals (CI). Statistical significance was set at P<.05. RESULTS Out of 354 peer reviewed publications that met the search criteria, a total of 9 relevant studies were identified and investigated the use of sodium carboxymethylcellulose (CMC) solutions, sodium hyaluronate/carboxymethylcellulose (HA/CMC) membranes, hyaluronate (HA), and fucoidan solutions. The odds of adhesions in horses treated with an adhesion barrier were significantly lower than untreated controls (OR=0.102; 95% CI [0.041, 0.254]; P<.001). When analyzed as subsets for each type of barrier, horses treated with HA/CMC membranes and CMC solutions had significant OR for fewer adhesions (OR=0.061; 95% CI [0.013, 0.292]; OR=0.119; 95% CI [0.034, 0.415], respectively; P<.001). CONCLUSION The meta-analysis demonstrated adhesion barriers provide a positive effect on the odds of adhesion formation. These results are tempered by the limitations of the study, including the small sample size and a bias towards publication of studies with only positive findings.

RECOGNITION OF AN IMPORTANT WATER QUALITY ISSUE AT ZOOS: PREVALENCE AND POTENTIAL THREAT OF TOXIC CYANOBACTERIA

Abstract: Zoo animals may be particularly vulnerable to water sources contaminated with cyanobacterial toxins given their nonvoluntary close association with this resource. However, the prevalence and potential threat of toxic cyanobacteria in this setting are unknown. Several otherwise unexplained yellow-bellied slider (Trachemys scripta scripta) deaths were documented in a zoo moat with recurring blooms of toxic Microcystis aeruginosa. Furthermore, an extremely high and potentially lethal concentration of the hepatotoxin microcystin (166 ng/g) was found in the liver of a necropsied turtle that died in this moat. A subsequent monthly survey of water quality revealed detectable concentrations of microcystin in all moats (0.0001 to 7.5 μg/L), with moats higher than 1 μg/L being significantly higher than the threshold for safe drinking water recommended by the World Health Organization. These results demonstrate that cyanobacterial blooms are an important water quality issue in zoos, and future research is necessary to identify potential associations among water quality, zoo animal health, and moat management strategies.

RESULTS OF THE MEGAVERTEBRATE ANALGESIA SURVEY: ELEPHANTS AND RHINO

Abstract An online survey utilizing Survey Monkey linked through the American Association of Zoo Veterinarians listserve examined current practices in megavertebrate analgesia. Data collected included drugs administered, dosing regimens, ease of administration, efficacy, and adverse events. Fifty-nine facilities (38 housing elephants, 33 housing rhinoceroses) responded. All facilities administered nonsteroidal anti-inflammatory drugs (NSAIDs), with phenylbutazone (0.25–10 mg/kg) and flunixin meglumine (0.2–4 mg/kg) being most common. Efficacy was reported as “good” to “excellent” for these medications. Opioids were administered to elephants (11 of 38) and rhinoceroses (7 of 33), with tramadol (0.5–3.0 mg/kg) and butorphanol (0.05–1.0 mg/kg) being most common. Tramadol efficacy scores were highly variable in both elephants and rhinoceroses. While drug choices were similar among institutions, substantial variability in dosing regimens and reported efficacy between and within facilities indicates the need for pharmacokinetic studies and standardized methods of analyzing response to treatment to establish dosing regimens and clinical trials to establish efficacy and safety.

Pyloric trichobezoar in a Canadian lynx (Lynx canadensis).

An adult female Canadian lynx (Lynx canadensis) was presented with a 3-wk history of anorexia and lethargy. Initial examination and diagnostics did not provide a diagnosis. The lynx continued to demonstrate vague clinical signs, including anorexia and an abnormal gait. During follow-up immobilizations 2 wk later, a barium gastrointestinal study revealed a pyloric obstruction. Abdominal exploratory surgery was elected, and a gastrotomy and an enterotomy of the proximal duodenum were performed to remove the pyloric obstruction. The obstruction was determined to be a trichobezoar. Fleas, a likely cause of hair ingestion through grooming, were noted during surgical preparation. The lynx made a full recovery from surgery. Reoccurrence of the trichobezoar was prevented after surgery with the use of monthly flea control and three times a week hairball laxative.

RESULTS OF THE MEGAVEREBRATE ANALGESIA SURVEY: GIRAFFE AND HIPPOPOTAMUS

Abstract Results of an online survey posted on the American Association of Zoo Veterinarians listserv examined the patterns of analgesic medication and pain management modalities used for captive giraffe and hippopotami. Compiled data included signalment, drugs administered, dosing regimens, subjective efficacy scores, ease of administration, and adverse events. Nineteen institutions exhibiting hippopotami (Hippopotamus amphibious) and pygmy hippopotami (Choeropsis liberiensis) and 45 exhibiting giraffe (Giraffa camelopardalis spp.) responded. Phenylbutazone was the most-commonly administered nonsteroidal anti-inflammatory drug (NSAID), followed by flunixin meglumine, but doses varied widely. Eight institutions reported adverse events from NSAID administration. Tramadol was the most-commonly administered opioid followed by butorphanol. Only one adverse event was reported for opioids. Twenty-three of 45 institutions exhibiting giraffe utilized alternative analgesia methods including gabapentin, glucosamine-chondroitin, local anesthetics, and low level laser therapy. Six of 19 institutions exhibiting hippopotami administered omega 3-6 fatty acids, gabapentin, glucosamine-chondroitin, and α-2 adrenergics to provide analgesia. While all reporting zoological institutions administered similar drugs, there was substantial variation and diversity in both dosing regimens and frequencies, indicating the need for both preclinical and clinical studies supporting dosing regimens.

RECURRENT EAR MITE (OTODECTES CYNOTIS) INFESTATION IN THREE RELATED GROUPS OF PATAGONIAN CAVIES (DOLICHOTIS PATAGONUM)

Abstract:  Two of three groups of Patagonian cavies (Dolichotis patagonum) contracted Otodectes cynotis infestations after exposure to mite-infested feral cats. Otodectes cynotis infestations were initially identified in 9 of 10 cavies in group 1. Multiple feral cats with O. cynotis infestations were observed in the vicinity of the Patagonian cavies and were subsequently removed. The Patagonian cavies were initially treated with ivermectin (0.4 mg/kg s.q.) every 2 wk, but ivermectin was discontinued after the third treatment due to injury to one of the Patagonian cavies during capture. Sixteen months after initial treatment, clinical signs of scratching the pinnae, hemorrhagic lesions on the ear margins, head shaking, and O. cynotis mites in the auricular canal were again noted in all Patagonian cavies in group 1. Repeated administration of ivermectin (0.4 mg/kg s.q. every 2 wk for three treatments) failed to eliminate the mites in two of the Patagonian cavies. Selamectin administered (20 mg/kg, topically between the shoulder blades) to all Patagonian cavies eliminated the mite infestation after a single application. The Patagonian cavies remained O. cynotis mite free for 2 yr, until males (group 2) and females (group 3) were separated for population control. Three months after separation, 8 of the 12 females in group 3 again were infestated with O. cynotis mites. Feral cats with O. cynotis infestations were again noted in the vicinity of group 3. A single dose of selamectin applied topically eliminated all mites in all treated Patagonian cavies. Group 2 was not exposed to feral cats and remained mite free.