Positive reinforcement conditioning as a tool for frequent minimally invasive blood and vaginal swab sampling in African lions (Panthera leo)

The South African National Research Foundation, the South African Veterinary Foundation and the Zebra Foundation for Veterinary Zoological Education.http://www.tandfonline.com/loi/haaw202021-12-29hj2020Anatomy and PhysiologyMammal Research Institut

Detection of Simonsiellaspp. in the vagina of lions and leopard in oestrus

Reports of the vaginal flora of wild cats such as lions or leopards are scarce. The micro‐organisms most commonly found in the vagina of clinically healthy cats are aerobic bacteria such as coagulase‐negative Staphylococcus, Streptococcus canis, and Escherichia coli. Simonsiellaspp are large Gram‐negative bacteria belonging to the Neisseriaceae family, typically found in the oral cavity and upper respiratory tract of many species. To date, there are no reports of the detection of Simonsiella spp in the vaginal flora of any felid. For a period of six months, daily behaviour monitoring was performed on six captive lionesses at a South African conservation centre, in parallel with the collection of vaginal swabs and interpretation of the resultant vaginal cytologies every other day. Oestrus was identified by typical female reproductive behaviours, as well as by enlarged and separated vulvar lips, and a predominant proportion of superficial cornified cells, clearing of the background, and high bacterial presence in the vaginal smear. Simonsiellaspp were identified by their characteristic morphology in 58% (60 of 103) of the vaginal samples collected during oestrus. They were also found in oral swabs of three out of three lions tested. Additionally, Simonsiellaspp were opportunistically found in a vaginal smear from a zoo housed female Sri Lankan leopard in oestrus, during a routine reproduction assessment. The finding of Simonsiella spp may be more common than previously suspected, transitory, and without detectable clinical relevance. A connection between occurrence of these bacteria and oestrus was apparent.The South African National Research Foundationhttp://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1439-05312019-12-01hj2018Anatomy and PhysiologyMammal Research Institut

Comparative reproduction of the female horse, elephant and rhinoceros: implications for advancing assisted reproductive technologies

Recent loss of rhinoceros subspecies has renewed interest in using more advanced assisted reproductive technologies (ART) in rhinoceroses and elephants. Currently, only semen collection, semen preservation and artificial insemination (AI) have been used repeatedly with success in these species. Although ovum pick-up (OPU) and intra-cytoplasmic sperm injection (ICSI) have been reported recently in rhinoceroses, the techniques are not yet optimised. In contrast, multiple ART applications are routinely used in the horse. Since elephants and rhinoceroses share some reproductive features with equids, we postulate that procedures such as OPU, ICSI, in vitro fertilisation (IVF) and embryo transfer (ET), which are well established in the horse, may represent a basis to develop protocols for endangered pachyderms. In this review, we summarise current knowledge on reproductive physiology relevant to ART. We discuss the current state of ART in all three families and the requirements for the successful implementation of OPU, ICSI, IVF and ET in these species

Use of gonadotropin releasing hormone (GnRH) vaccines for behavioural and reproductive control in managed Asian elephant Elephas maximus and African elephant Loxodonta africana populations

Because of their size and reproductive anatomy, surgical castration is not a practical option in adult elephants. However, similar to other species in human care, the effects of gonadectomy are desired in specific situations. This may be for contraceptive purposes, or for behavioural or veterinary management of elephants in human care or wild elephants managed in small reserves. Research into non‐surgical contraceptive measures for wild and domestic animals has resulted in an array of hormonal and immunological options to downregulate gonadal function. Driven by the production‐animal industry, commercial gonadotrophin releasing hormone (GnRH) vaccines are readily and cheaply available. This immunocontraceptive is effective in both males and females as it inhibits gonadotrophic hormone release and, thus, downstream stimulation of testicles and ovaries. Here, available studies from the past 10 years on the GnRH vaccine application in male and female elephants are reviewed. Furthermore, we add our own experience gathered from the treatment of male African elephants Loxodonta africana and female Asian elephants Elephas maximus in human care. The GnRH vaccine offers a viable approach for various management purposes. It should be noted that the GnRH vaccine was not primarily designed as a reversible contraceptive. Therefore, its use must be well justified on an individual basis and the effects closely monitored.https://zslpublications.onlinelibrary.wiley.com/journal/174810902020-07-01hj2020Anatomy and PhysiologyProduction Animal Studie

Cardiopulmonary parameters and arterial blood gases during etorphine-medetomidine-midazolam immobilization in free-ranging black rhinoceroses (Diceros bicornis) undergoing electro-ejaculation : a preliminary study

Conservation management interventions for the critically endangered black rhinoceros (Diceros bicornis) require immobilization, which offer opportunities for semen collection and cryopreservation to establish genetic reservoirs. In free-ranging rhinoceroses, a combination of the potent opioid etorphine and the tranquilizer azaperone is routinely used for chemical immobilization but is associated with muscle rigidity and severe cardiopulmonary changes. Additionally, azaperone inhibits semen emission. Seven free-ranging, male, sexually mature black rhinoceroses were immobilized with an alternative protocol consisting of 4.5mg etorphine, 5mg medetomidine, 50mg midazolam and 2,500 IU hyaluronidase delivered remotely by darting from a helicopter. During the immobilization, electro-ejaculation was performed with a portable electro-ejaculator, and a species-specific rectal probe. Animals were observed for muscle tremors. Longitudinal changes in respiratory rate, heart rate and peripheral oxyhemoglobin saturation, measured at 5min intervals, were assessed using a general mixed model. Non-invasive oscillometric blood pressure and arterial blood gas variables were measured at first handling and before reversal and compared using the Wilcoxon rank sum test. All animals were successfully immobilized, showed no muscle tremors, presented with normal heart rates and lactate concentration (<5 mmol/L), recovered uneventfully, but experienced acidemia, hypoxemia and hypercapnia. Induction time and total time in recumbency were 4.2 ± 0.41 and 38.4 ± 6.9min, respectively. Electro-stimulation commenced after 11.7 ± 3.98min and completed after 24.3 ± 6.65min. Semen-rich fractions were successfully collected from six animals. Our observations indicate that etorphinemedetomidine- midazolam provides a promising immobilization protocol for free-ranging black rhinoceroses, that allows for successful electro-ejaculation.Rhino Force SA NPChttps://www.frontiersin.org/journals/veterinary-science#am2022Production Animal Studie

Arterial blood gases and cardiorespiratory parameters in etorphine-medetomidine-midazolam immobilized free-ranging and game-farmed Southern white Rhinoceroses (Ceratotherium simum simum) undergoing electro-ejaculation

With the rapid loss of individuals in the wild, semen cryopreservation has gained importance to safeguard the genetic diversity of white rhinoceroses (Ceratotherium simum). For semen collection via electro-ejaculation, immobilization of free-ranging individuals requires the potent opioid etorphine, which is routinely combined with azaperone, but causes hypoxemia, hypercarbia, acidemia, muscle rigidity, tachycardia, and systemic hypertension. In this study, the suitability of two alternative immobilization protocols including etorphine, medetomidine, and midazolam at different doses (high vs. low etorphine) was evaluated in adult white rhinoceros bulls in two different management systems (free-ranging vs. game-farmed) and undergoing electro-ejaculation. Fourteen free-ranging (Group 1) and 28 game-farmed rhinoceroses (Group 2) were immobilized with ≈2.5 µg/kg etorphine (high dose), ≈2.5 µg/kg medetomidine, ≈25 µg/kg midazolam and 1,500–1,700 IU hyaluronidase and received ≈2.5 µg/kg of butorphanol intravenously at first handling. Twenty game-farmed animals (Group 3) received ≈1 µg/kg etorphine (low dose), ≈5 µg/kg medetomidine, ≈25 µg/kg midazolam and 1,700 IU hyaluronidase. Respiratory rate, heart rate and peripheral hemoglobin oxygen saturation (SpO2) were measured at 5-min intervals; non-invasive oscillometric blood pressures and arterial blood gases at first handling and before reversal of the immobilization; serum clinical chemistry analytes and hematocrit at first handling. Generalized mixed models (fixed factors: group, time, recumbency; random factor: individual rhinoceros) were applied to compare longitudinal changes between free-ranging and game-farmed rhinoceroses immobilized with the higher etorphine dose (Groups 1 and 2), and between the two protocols tested in the game-farmed rhinoceroses (Groups 2 and 3). All animals were successfully immobilized, presented with normal lactate concentrations (<5 mmol/L), experienced no muscle tremors and recovered uneventfully. Hypoxemia and hypertension persisted throughout the immobilization in all groups. Acidemia and hypercarbia were absent in Group 1, but present in the game-farmed animals. The lower etorphine dose in Group 3 resulted in significantly longer induction times, however, tachycardia was not observed. SpO2 was higher for sternal vs. lateral recumbency. Semen-rich fractions were recovered following electro-stimulation in 46 out of the 62 animals. Our findings suggest that etorphine-medetomidine-midazolam provides effective immobilization with fewer side effects compared to previous reports in white rhinoceroses and is suitable for successful electro-ejaculation.Rhino Force SA NPChttps://www.frontiersin.org/journals/veterinary-sciencedm2022Mammal Research InstituteProduction Animal Studie

Effects of GnRH vaccination in wild and captive African Elephant bulls (Loxodonta africana) on reproductive organs and semen quality

OBJECTIVES: Although the African elephant (Loxodonta africana) is classified as endangered by the International Union for Conservation of Nature (IUCN), in some isolated habitats in southern Africa, contraception is of major interest due to local overpopulation. GnRH vaccination has been promoted as a non-invasive contraceptive measure for population management of overabundant wildlife. We tested the efficacy of this treatment for fertility control in elephant bulls. METHODS: In total, 17 male African elephants that were treated with a GnRH vaccine were examined in two groups. In the prospective study group 1 (n = 11 bulls, ages: 8±36 years), semen quality, the testes, seminal vesicles, ampullae and prostate, which were all measured by means of transrectal ultrasound, and faecal androgen metabolite concentrations were monitored over a three-year period. Each bull in the prospective study received 5 ml of Improvac® (1000 μg GnRH conjugate) intramuscularly after the first examination, followed by a booster six weeks later and thereafter every 5±7 months. In a retrospective study group (group 2, n = 6, ages: 19±33 years), one examination was performed on bulls which had been treated with GnRH vaccine for 5±11 years. RESULTS: In all bulls of group 1, testicular and accessory sex gland sizes decreased significantly after the third vaccination. In six males examined prior to vaccination and again after more than five vaccinations, the testis size was reduced by 57.5%. Mean testicular height and length decreased from 13.3 ± 2.6 cm x 15.2 ± 2.8 cm at the beginning to 7.6 ± 2.1 cm x 10.2 ± 1.8 cm at the end of the study. Post pubertal bulls (>9 years, n = 6) examined prior to vaccination produced ejaculates with viable spermatozoa (volume: 8±175 ml, sperm concentration: 410-4000x106/ml, total motility: 0±90%), while after 5±8 injections, only 50% of these bulls produced ejaculates with a small number of immotile spermatozoa. The ejaculates of group 2 bulls (vaccinated >8 times) were devoid of spermatozoa. Faecal androgen metabolite concentrations measured in captive males decreased significantly after the fourth vaccination. None of the males entered musth during the treatment period. CONCLUSIONS: Our results showed a marked decrease in semen quality, testicle and secondary sex gland sizes following repeated GnRH vaccinations. After 2±4 years of continuous treatment every 5±7 months, the effects were similar to surgical castration.ISIScopu

Raw data ELE GnrH.xlsx

<p>Effects of GnRH Vaccination in Wild and Captive African Elephant Bulls (<i>Loxodonta africana</i>) on Reproductive Organs and Semen Quality<b>.</b></p><p><b>PLOSone Publication</b></p><p>The table added here show the raw data collected.</p><p><br></p

Konstruktion eines Maulgatters zur Untersuchung der Maulhöhle, Durchführung von Bronchoskopie und Sondierung des Magens bei Elefanten

OBJECTIVE : Here we tested the application of a full mouth speculum to sedated elephants in human care to gain access to the oral cavity, the trachea (bronchi) and esophagus (stomach) and therefore improve diagnostic and therapeutic options in elephant medicine. The construction of this oral speculum for elephants and the procedure are described. MATERIALS AND METHODS : The oral speculum is a steel construction consisting of 2 bite plates of 0.8 × 60.0 × 8.0 cm attached between 2 threaded guiding poles (40 cm). Through crank handles, the metal plates are dispersed once placed between the elephant‘s jaws in front of the molars. The oral speculum was applied in 26 elephants (6,16 Asian elephants, and 1,3 African elephants) during standing sedation. RESULTS : All sedated elephants tolerated the positioning of the mouth opener and subsequent manipulations well. The mouth opener was applied for the following procedures: inspection of the oral cavity (n = 2), placing a stomach tube (n = 16), and/or performing endoscopic examinations such as bronchoscopy (n = 20) and/or gastroscopy (n = 8). CONCLUSION : This method provides a new possibility to open the jaws to gain access to the molars, larynx and pharynx in captive elephants without full immobilization. Valuable samples for diagnostics may be obtained or animals medicated via stomach tube with this application. CLINICAL RELEVANCE : The mouth opener provides veterinarians with a new option to perform necessary diagnostic and therapeutic procedures around the oral cavity, airways and stomach in captive elephants during standing sedation with no need for a full anaesthesia.GEGENSTAND UND ZIEL : Ziel der Untersuchungen war, den Einsatz eines Maulgatters für Elefanten zu erproben. Der Artikel beschreibt die Konstruktion des Maulgatters sowie Anwendungsmöglichkeiten am sedierten Elefanten zur Spreizung der Kiefer, um den Zugang zu Maulhöhle, Trachea (Bronchien) und Ösophagus (Magen) für diagnostische und therapeutische Zwecke zu ermöglichen. MATERIAL UND METHODEN : Das Maulgatter ist eine stabile Stahlkonstruktion bestehend aus 2 Flacheisen von 0,8 × 60,0 × 8,0 cm Größe, die über 2 Gewindestangen von 40 cm Länge verbunden sind. Über 2 Handkurbeln können die Gewindestangen gedreht und somit die Flacheisen auseinander bewegt werden. Das Maulgatter wurde an 26 Elefanten (1,3 Loxodonta africana und 6,16 Elephas maximus) mit unterschiedlichen Anwendungen getestet. ERGEBNISSE : Bei insgesamt 26 Elefanten ließ sich das Maulgatter unter Standsedation erfolgreich einsetzen, um die Maulhöhle zu kontrollieren (n = 2), eine Magenschlundsonde zu legen (n = 12) und/oder eine Broncho- (n = 20) oder Gastroskopie (n = 8) durchzuführen. SCHLUSSFOLGERUNG : Durch die besondere Anatomie des Elefanten sind Ösophagus und Trachea nicht wie beim Pferd einfach über die Nasenöffnung zu erreichen. Das Maulgatter ermöglicht auf unkomplizierte Weise am stehenden sedierten Tier einen direkten Zugang, um Endoskopien durchzuführen, Proben zu nehmen oder Medikamante zu verabreichen. KLINISCHE RELEVANZ : Die beschriebene Methode ist eine neue Option für diagnostische und therapeutische Maßnahmen im Bereich von Maulhöhle, Atemwegen, Ösophagus und Magen beim Elefanten ohne Vollnarkose.https://www.thieme.de/de/tieraerztliche-praxis-grosstiere/aktuelle-ausgabe-120139.htm2022-11-17hj2022Mammal Research Institut

Use of anti-gonadotropin-releasing hormone vaccines in African elephants (Loxodonta africana): A review

Background: Androgen-related aggressive behaviour and musth cause serious problems in captive African elephant bulls and often lead to human and animal injuries, and damage to property. Objectives: To review the work carried out with anti-gonadotropin-releasing hormone (GnRH) vaccines to control androgen-related behaviour and fertility in captive and free-ranging elephant bulls and the induction of anoestrus in elephant cows. Method: In the first study, an anti-GnRH vaccine from Pepscan was tested in six bulls (four captive and two free-ranging). Once the vaccine Improvac® became available, the effect on behaviour, the reproductive organs and semen quality was tested. Improvac® was also used to attempt induction of anoestrus in elephant cows. Results: The first study proved that aggressive behaviours are significantly associated with increased faecal androgen concentrations. Musth (n = 1) and aggressive behaviour (n = 2) were down regulated and correlated with a decline in faecal androgen concentrations. Aggression and musth could be controlled with Improvac® (600 µg), but were more consistent when the dose was increased to 1000 µg administered every five to six months. The same dose down regulated testicular function and bulls (n = 17) were rendered infertile within 12 months after commencement of treatment. Initial attempts to induce anoestrous with 600 µg in free-ranging elephant cows gave inconclusive results, but 1000 µg in captive cows delivered five-monthly was successful. Conclusion: The treatment of elephant bulls with Improvac® resulted in the successful down-regulation of androgen-related behaviour and sperm production in captive and wild elephant bulls of various ages (≤ 34 years). Preliminary studies to induce anoestrus in cows with Improvac® appear to be successful