Female long-term sperm storage results in viable offspring in the Himalayan Mountain Pitviper, Ovophis monticola.

The ability of females to store sperm for extended periods in their reproductive tracts (termed long-term sperm storage, LTSS) has been reported across a diversity of vertebrate taxa. The evolutionary, ecological, and physiological significance of LTSS is wide-ranging and includes the ability to produce offspring when mates may be temporally scarce by way of decoupling copulation from ovulation, inbreeding avoidance, and the generation and maintenance of genetic diversity in progeny. Among vertebrate lineages, nonavian reptiles exhibit a remarkable capacity for LTSS, with the production of viable offspring reported after periods exceeding 6 years since prior contact with a potential mate. Given that female reptiles are able to store viable sperm for prolonged periods, it is important to disentangle LTSS from that of facultative parthenogenesis (FP), a reproductive trait which appears widespread among all reptile lineages. The implications of this distinction are particularly important in the context of the development and management of captive breeding programs. To accurately determine between the two reproductive strategies, genomic screening is highly recommended. Following a period of isolation for 13 months from a potential male mate, a female Himalayan Mountain Pitviper (Ovophis monticola) produced a clutch of three male offspring. Here, through genome-scale analyses of the female and her progeny, we document the first record of LTSS in this genus and exclude FP as the alternative hypothesis.

Radiographic and sonographic features, and histogram parameters of the liver and spleen in healthy Toco toucans (Ramphastos toco, Müller 1976).

This study aimed to evaluate radiographic and sonographic features, and histogram parameters based on grayscale ultrasound of the liver and spleen in healthy toco toucans. Fifteen adult toco toucans (Ramphastos toco), seven females and eight males, weighing approximately 650 g, were enrolled in the study. On the right lateral radiographic view, the liver was visualized in the midventral region of the coelomic cavity; ultrasonographically, the liver was located in the middle portion of the coelomic cavity in close relationship to the heart, and thoracic and abdominal air sacs. Two hepatic lobes were identified; the right lobe was larger than the left one. The spleen was visualized in 10 toco toucans on radiographs and only in eight toucans on ultrasound exams. The gallbladder was identified only on ultrasound. On the right lateral radiographic view, the spleen was visualized dorsal to the proventriculus and ventral to the lungs/air sacs as an oval shape. Ultrasonographically, the spleen was observed caudal to the liver, cranial to the proventriculus and craniodorsal to the ventriculus. There was no significant difference in spleen length for either radiographic or ultrasound measurements. The brightness intensity (BI) for mean gray level (G) and standard deviation of gray levels (SG ) in the liver were 101.315 (± 16.170) and 12.453 (± 2.616), respectively. Mean G and SG levels in the spleen were 63.940 (± 18.321) and 7.494 (± 3.595), respectively. In conclusion, the sonographic features and histogram parameters indicated that the liver is more echogenic and heterogeneous than the spleen, which must be considered for diagnosing alterations in these organs.

Serum vitamin D in sanctuary chimpanzees (Pan troglodytes) in range countries: A pilot study.

BACKGROUND: Vitamin D is essential for skeletal health, calcium homeostasis and general health. The major and more stable form of vitamin D in circulation is 25-hydroxyvitamin D (25-OH-D); this is the most valuable indicator of vitamin D status. There are studies on laboratory and zoo-housed chimpanzees; however, serum vitamin D status has not been documented in chimpanzees in range countries. OBJECTIVES: (1) Determine the range of circulating 25-OH-D concentrations in chimpanzees in range countries. (2) Assess the influence of age, sex, and sun exposure on 25-OH-D serum concentrations. METHODS: Opportunistic blood samples were obtained from 127 clinically healthy chimpanzees. Serum 25-OH-D concentration was measured with a commercially available competitive ELISA. RESULTS: The median overall 25-OH-D concentration for chimpanzees in range countries was 46.24 nmol/L (range: 17.10-109.23 nmol/L). Males had a significantly lower concentration (40.15 nmol/L) than females (49.61 nmol/L), and infants (37.99 nmol/L) had a significantly lower concentration than adults (46.04 nmol/L). Concentrations of 25-OH-D in chimpanzees in sunnier habitats were significantly higher compared to thick tropical forest habitat. CONCLUSION: The present constitutes a large dataset of serum 25-OH-D concentrations in range country sanctuary chimpanzees and contributes to document normal ranges. Age, sex, and sun exposure influenced serum concentrations of 25-OH-D in sanctuary chimpanzees.

Partial Amputation of a Nonreducing Prolapsed Phallus in an Ostrich (Struthio camelus).

A 19-year-old male ostrich (Struthio camelus) was referred to a veterinary teaching hospital (São Paulo State University, Botucatu, Brazil) due to a 6-month history of recurrent prolapse of the phallus. On physical examination, 2 ulcerative wounds were present on the phallus, as well as caseous plaques and myiasis. Conservative treatment resulted in improvement but prolapse of the phallus remained. Thus, a decision was made to perform a partial phallectomy. The surgery was successful and no postoperative complications occurred. When the ostrich was reexamined 6 months postsurgery, the ostrich was alert, in good health, and the surgical site completely healed. The owner verbally reported no recurrence of the phallus prolapse 1 year after surgery.

Serial ultrasonographic measurements of fetal parameters over three successive pregnancies in a captive Eastern black-and-white colobus monkey (Colobus guereza).

This study provides ultrasonographic fetal growth charts for the Eastern black-and-white colobus monkey (Colobus guereza). Throughout three consecutive gestations (-162 to -2 days to parturition) in a single dam, we opportunistically obtained ultrasonographic measurements for the following parameters: biparietal diameter, head circumference, humerus length, femur length, tibia length, radius length, thoracic width, kidney length, and crown-rump length. Biparietal diameter was the most consistently measured parameter. First detection of fetuses occurred between 96 and 162 days before parturition. This report demonstrates that voluntary transabdominal ultrasound can be well-tolerated in the colobus monkey using operant conditioning. These findings may be useful to assess fetal development and predict parturition dates in the absence of a known conception date in this species.

Radiographic measurements of the hind limbs in capuchin monkeys (Sapajus spp.).

BACKGROUND: Radiographic measurements are a useful tool to assess skeletal health and development in primates. This study aimed to perform radiographic measurements of hind limbs in capuchin monkeys. METHODS: Twelve Sapajus spp. were used; ten adults and two sub-adults, nine females and three males. RESULTS AND CONCLUSIONS: The pelvimetry showed that the pelvic inlet area presented mean values of 7.63, 10.23, and 5.43 cm2 for adult males, adult females, and sub-adult females, respectively. The mean inclination angle was 129.45° and the mean values for the mechanical lateral proximal and distal femoral angles were 102.32° and 90.93°, respectively. The mean values for the anatomic lateral proximal and distal femoral angles were 104.59° and 85.98°, respectively. In conclusion, the radiographic measurements proved feasible for hind limb evaluation in Sapajus spp. and can be used for comparison with animals presenting orthopedic lesions.

MORBIDITY AND MORTALITY OF BINTURONGS (ARCTICTIS BINTURONG) IN UNITED STATES ZOOLOGICAL INSTITUTIONS (1986-2019).

Veterinary literature on diseases in binturong (Arctictis binturong) is often limited to case series or reports without population-level information. Morbidity and mortality data were collected from North American institutions through survey completion or submission of medical records. Responses included information about 74 individuals (37 males, 30 females, and 7 unknown neonates) representing 22 institutions, from 1986 to 2019. Antemortem data were available from 39 individuals, and postmortem data were available from 53 individuals. Eighteen individuals had both antemortem and postmortem records available. Age (mean ± SD) at death for adults (n = 41) was 15.2 ± 4.3 yr. Morbidity events were summarized by affected organ system; 160 events were reported. The systems with the most frequently reported events were gastrointestinal (33%; 53/160), integumentary (19%; 31/160), urinary (12%; 20/160), and musculoskeletal (19 [12%] of 160). Excluding neonates, primary causes of mortality included neoplasia (51%; 21/41), infectious or inflammatory diseases (24%; 10/41), and cardiovascular disease (17%; 7/41). Neoplasms (51%; 21/41) confirmed on histopathology included renal adenocarcinoma (47%; 10/21); mammary carcinoma (14%; 3/21); pancreatic islet cell carcinoma (2 [10%] of 21); and single instances of multicentric lymphoma, uterine carcinoma, and submucosal urethral adenoma. There were three additional cases of presumed neoplasia without histopathologic confirmation; masses were detected in the liver, heart base, and pancreas. Metastases were reported in 15 (71%) of 21 neoplasms. Although neoplasia and cardiovascular disease were common causes of mortality, they were rarely diagnosed antemortem. Neoplasia was often malignant and generally diagnosed after metastasis. Preventive medicine protocols with improved renal and cardiovascular evaluation are warranted and may result in earlier detection of subclinical disease in binturong.

Nighttime behavior and the length of social relationships in male Asian elephants.

To evaluate elephant welfare, it is important to understand their use of time both during day and night. The length of social relationships can influence how much time they spend in different activities. We assessed daytime and nighttime activity budgets of male Asian elephants at Denver Zoo and examined how length of relationships influenced nighttime behavior. Using scan sampling we investigated activity budget and proximity to a conspecific, and used General Estimating Equations to compare them across day and night and across new and established dyads at night. During daytime, elephants spent significantly more time exhibiting affiliative and agonistic behaviors, and in proximity to a conspecific, and less time resting, compared to night. Overnight, the odds of resting were significantly lower in new social dyads compared to established dyads, and new dyads spent more time exhibiting agonistic behavior and in proximity to a conspecific compared to established dyads. Our study suggests that male elephants at Denver Zoo have developed strong relationships and highlights the importance of systematically observing elephants overnight so that managers make decisions that improve animal welfare.

Social Dynamics of a Newly Integrated Bachelor Group of Asian Elephants (Elephas maximus): Welfare implications.

Male associations are a typical component of elephant society, allowing bulls to practice appropriate social behavior. To improve zoo elephant welfare, it is important to provide bulls with social opportunities. In fall 2018, Denver Zoo added two bull Asian elephants to its existing all-male group of three bulls, offering the opportunity to conduct a systematic behavioral study of the integration of the new bulls into the resident group. We recorded aggressive and affiliative behaviors before, during, and after the introduction of the new males. The proportion of aggressive behavior was significantly higher during the five-month introduction period compared to before their introduction. By the end of the study period, the elephants engaged in significantly more affiliative behavior and less aggressive behavior than during the introduction period, suggesting they had formed a new stable social dynamic. These results suggest group compatibility and positive elephant welfare resulting from housing male elephants together and can be used to inform management plans for bull elephants that prioritize their welfare.

Integrated approaches to howler monkey (Alouatta spp.) medicine in professional care and conservation.

Howler monkeys (Alouatta spp.) are threatened by anthropogenic pressures such as habitat fragmentation and deforestation, while conservation efforts are challenging to coordinate as natural geographic distribution ranges are the largest of any New World primate. On a One Health front, howler monkeys represent a great model to investigate the infectious disease dynamics between wild primates and humans as several infectious diseases affecting howlers have a demonstrated zoonotic potential. Howler monkey populations in professional care offer a window to investigate susceptibility to diseases in this species such as yellow fever (YF) and malaria, plus a myriad of endoparasite phyla, as well as vector-borne diseases such as Chagas disease and leishmaniasis. More studies are urgently needed to provide species-specific, medically relevant information as well as clinical descriptions of animals considered medically healthy. Moreover, howler monkeys are a challenging species to breed and maintain in professional care; additionally, reproductive parameters have been published only for a handful of species in this genus. On a One Health approach communication and collaborative health surveillance involving wildlife and zoo experts will ease the identification of factors that contribute to disease emergence facilitating the integration of human, animal, and environmental health. The One Welfare concept is based on the inextricable connection among animal welfare, human welfare, and environmental conservation. Integrating One Health and One Welfare into actions both in-situ and ex-situ will promote the sustainability of the forests and restoration of the ecosystems that those species inhabit, transitioning to a comprehensive One Conservation approach.

Of Elephants and Other Mammals: A Comparative Review of Reproductive Tumors and Potential Impact on Conservation.

Reproductive tumors can impact conception, pregnancy, and birth in mammals. These impacts are well documented in humans, while data in other mammals are limited. An urgent need exists to understand the reproductive impact of these lesions in endangered species, because some endangered species have a documented high prevalence of reproductive tumors. This article documents that the prevalence of both benign and malignant neoplasia differs between African and Asian elephants, with Asian elephants more frequently diagnosed and negatively affected by both. The prevalence of these tumors across mammalian species is compared, and impact plus treatment options in human medicine are reviewed to inform decision making in elephants. Evidence suggests that reproductive tumors can negatively impact elephant conservation. Future studies that document reproductive outcomes, including the success of various treatment approaches in elephants with tumors will benefit conservation efforts.

Vaginal foreign bodies in six nonhuman primates with underlying pathological conditions.

Four female Japanese macaques (Macaca fuscata) from the same group as well as Wolf's guenon (Cercopithecus wolfi) and a Western lowland gorilla (Gorilla gorilla gorilla) from a second institution presented with vaginal foreign bodies in parallel with diseases of the urogenital tract or with endocrine disorders. These foreign bodies were associated with a mild to marked, diffuse vaginitis in all cases. Underlying pathological conditions consisted of a cavernous uterine hemangioma in a 20-year-old macaque, diffuse endometritis in a 21-year-old macaque, an in situ endometrial carcinoma in a 24-year-old macaque, endometritis and an ovarian cyst-like structure in the 27-year-old Western lowland gorilla, chronic cystitis and chronic renal disease in a 24-year-old macaque, and a history of hypothyroidism with irregular reproductive cycles in the 12-year-old Wolf's guenon. Vaginal foreign bodies have been reported in nonhuman primates used in biomedical research, but their concurrence with underlying conditions has not been explored. In women, vaginal foreign bodies have been linked to serious underlying medical conditions. This case series emphasizes the recommendation to investigate this abnormal behavior for underlying medical or adverse psychosocial conditions in primates under human care.

A Multi-Institutional Collaboration to Understand Neoplasia, Treatment and Survival of Snakes.

This multi-institutional collaborative study of neoplasia in snakes reviewed medical records of snakes at each facility to determine species prevalence, survival, and methods of treatment. Complete species numbers of snakes were also collected at each facility. In total, 65 species, 133 snakes, and 149 unique neoplasias were included in this study. Affected species, age, sex, and their tumor prevalence, tumor type and location, metastasis, treatment, and survival data are reported. The highest species-specific tumor prevalence was in Common or Northern Watersnakes (Nerodia sipedon) (30.8%, n = 4 of 13), Eastern Diamond-Backed Rattlesnakes (Crotalus adamanteus) (26.3%, n = 5 of 19), and Timber rattlesnakes (Crotalus horridus) (22.7%, n = 5 of 22). Malignant tumors predominated (86.6%, n = 129 of 149) with soft tissue sarcomas being the most common (30.2%, n = 45 of 149). Snakes with malignant neoplasia, metastases, or indeterminate presence of metastases were statistically more likely to die from their neoplasms than snakes having either benign neoplasia or no diagnosed metastases (p < 0.05). Gender, taxonomic family, and species of those evaluated did not significantly affect the outcome of snakes with neoplasia. Only 27.1% (n = 36 of 133) of snakes received a reported form of treatment and, for those treated, surgical excision was the most common treatment modality. There was not a significant difference in outcome based on treatment; however, surgery and chemotherapy were associated with death from a cause other than their tumor.

Reproductive one health in primates.

One Health is a collaborative trans-disciplinary approach to health; integrating human, animal, and environmental health. The focus is often on infection disease transmission and disease risk mitigation. However, One Health also includes the multidisciplinary and comparative approach to disease investigation and health of humans, animals, and the environment. One key aspect of environmental/ecosystem health is conservation, the maintenance of healthy, actively reproducing wildlife populations. Reproduction and reproductive health are an integral part of the One Health approach: the comparative aspects of reproduction can inform conservation policies or breeding strategies (in situ and ex situ) in addition to physiology and disease. Differences in reproductive strategies affect the impact poaching and habitat disruption might have on a given population, as well as ex situ breeding programs and the management of zoo and sanctuary populations. Much is known about chimpanzees, macaques, and marmosets as these are common animal models, but there is much that remains unknown regarding reproduction in many other primates. Examining the similarities and differences between and within taxonomic groups allows reasonable extrapolation for decision-making when there are knowledge gaps. For example: (1) knowing that a species has very low reproductive rates adds urgency to conservation policy for that region or species; (2) identifying species with short or absent lactation anestrus allows ex situ institutions to better plan contraception options for specific individuals or prepare for the immediate next pregnancy; (3) recognizing that progestin contraceptives are effective contraceptives, but may be associated with endometrial hyperplasia in some species (in Lemuridae but not great apes) better guides empirical contraceptive choice; (4) recognizing the variable endometriosis prevalence across taxa improves preventive medicine programs. A summary of anatomical variation, endocrinology, contraception, pathology, and diagnostics is provided to illustrate these features and aid in routine physical and postmortem examinations as well as primate management.

Reproductive suppression of giraffe (Giraffa camelopardalis) under managed care using a GnRH immunological product.

Giraffe present unique contraception challenges as males persistently pursue females during estrus. Year-round pursuit during frequent recurring estrus can pose significant risk under slippery conditions. Complete ovarian suppression is a useful tool in giraffe because it eliminates estrous behavior, interest from the male, and controls reproduction. Effective reproduction control in giraffes has been achieved with porcine zona pellucida, oral melengestrol acetate, and depot medroxy-progesterone acetate. However, these methods allow some degree of folliculogenesis and estrous behavior. Improvest® is a gonadotropin releasing hormone (GnRH) immunological product that elicits antibodies against GnRH and abrogates the effects of endogenous GnRH. This study evaluated the efficacy of Improvest® for gonadal suppression in seven females and one male giraffe by monitoring steroid hormones. Seven female giraffe were treated intramuscularly with an initial dose, a booster at 4 weeks and maintenance boosters at 3-month intervals (600 µg/dose) for 12 months. Six females were on supplemental contraception during the induction phase because separation from males was not possible. In the male (treated with 400 µg), testosterone concentrations decreased after the second injection. However, even with low serum testosterone concentrations, mounting (of nontreated females) behavior was still observed occasionally. Ovarian activity was suppressed in all treated females and interest by the males stopped; supplemental contraceptives (during the induction phase) did not impede the effect of Improvest®. After 15.3 months (seven doses), Improvest® was discontinued in three females which no longer needed contraception. In these females, ovarian activity was noted approximately 90 days after the last dose.

Host Identity and Geographic Location Significantly Affect Gastrointestinal Microbial Richness and Diversity in Western Lowland Gorillas (Gorilla gorilla gorilla) under Human Care.

The last few decades have seen an outpouring of gastrointestinal (GI) microbiome studies across diverse host species. Studies have ranged from assessments of GI microbial richness and diversity to classification of novel microbial lineages. Assessments of the "normal" state of the GI microbiome composition across multiple host species has gained increasing importance for distinguishing healthy versus diseased states. This study aimed to determine baselines and trends over time to establish "typical" patterns of GI microbial richness and diversity, as well as inter-individual variation, in three populations of western lowland gorillas (Gorilla gorilla gorilla) under human care at three zoological institutions in North America. Fecal samples were collected from 19 western lowland gorillas every two weeks for seven months (n = 248). Host identity and host institution significantly affected GI microbiome community composition (p < 0.05), although host identity had the most consistent and significant effect on richness (p = 0.03) and Shannon diversity (p = 0.004) across institutions. Significant changes in microbial abundance over time were observed only at Denver Zoo (p < 0.05). Our results suggest that individuality contributes to most of the observed GI microbiome variation in the study populations. Our results also showed no significant changes in any individual's microbial richness or Shannon diversity during the 7-month study period. While some microbial taxa (Prevotella, Prevotellaceae and Ruminococcaceae) were detected in all gorillas at varying levels, determining individual baselines for microbial composition comparisons may be the most useful diagnostic tool for optimizing non-human primate health under human care.

Ultrasonography of the gastrointestinal tract of toco toucans (Ramphastos toco).

Baseline data for wall thickness in healthy toucans can be used to diagnose disease in sick individuals. This study aimed to evaluate the use of ultrasonography for the examination of the gastrointestinal tract of healthy toco toucans (Ramphastos toco). Ten non-anaesthetized healthy adult toco toucans were evaluated. Food was withheld for 3 h prior to the ultrasound examination. Longitudinal and transverse images of the gastrointestinal tract were obtained using ventromedian cranial and ventromedian caudal approaches to evaluate the morphology and stratification. The wall thickness of the sections was measured: proventriculus and ventriculus, duodenum, jejunum/ileum, and cloaca. Additionally, the thickness of the mucosal layer of the duodenum and jejunum/ileum was also measured. Subsequently, the ratios of the duodenal wall thickness to the duodenal mucosa layer and the total wall thickness of the jejunum/ileum to the jejunum/ileum layer mucosa were stablished. The wall thickness of the proventriculus was significantly higher than the wall thickness of ventriculus. The wall thickness of the duodenum was significantly higher than the wall thickness of jejunum/ileum. The mean ratios of the duodenal wall thickness to the duodenal mucosa layer and the wall thickness of the jejunum/ileum to the jejunum/ileum layer were 83.15% and 80.07%, respectively. In conclusion, it was possible to assess the stomach and intestines using ultrasound examination in healthy toco toucans, due to their medium body size and no interference by surrounding air sacs.

HYDROMETRA IN ROCK HYRAX (PROCAVIA CAPENSIS) UNDER MANAGED CARE.

In zoos, rock hyrax (Procavia capensis) are commonly maintained in groups and population control is necessary. Here we report on hydrometra prevalence in a group of rock hyraxes. Prevalence of hydrometra in this small group (4/8) far exceeds reproductive pathology prevalence previously reported for this species under managed care. Affected females were nulliparous, but had not been contracepted; instead they were maintained as a single-sex group. The first case presented as sudden death and three additional cases were diagnosed antemortem via ultrasound. Two of these underwent ovariohysterectomy to treat the severe hydrometra. The last case was a mild hydrometra and during follow-up exam was found to have spontaneously resolved. Detailed information regarding clinical presentation, diagnostics and surgical techniques are provided.

Severe Hydroxyapatite Deposition Disease in Binturongs (Arctictis binturong).

In a collection of 6 young binturongs (Arctictis binturong), 3 presented with anorexia, lethargy, and footpad swelling diagnosed by surgical biopsy as calcinosis circumscripta. Despite supportive care over the next 4 years, affected binturongs had progressive weight loss, hyperphosphatemia, and hyperkalemia and developed large radiodense deposits in tissues adjacent to appendicular joints, thoracolumbar vertebrae, and the sternum. Two binturongs died and necropsies showed severe periarticular mineral deposition with fibrosis and granulomatous inflammation. Additionally, binturongs had mineralization of distal renal tubules, mineralization of the renal interstitium, and marked mineralization of the pulmonary interstitium and peribronchiolar smooth muscle. Foot biopsies from the surviving binturong were evaluated by spectroscopic microanalysis and were positive for hydroxyapatite, Ca10(PO4)6(OH)2. Periarticular and footpad mineralization in these binturongs is consistent with hydroxyapatite deposition disease, which is rarely described in humans and animals as a primary familial condition or a secondary condition often associated with renal injury.

Diagnosis and treatment of endometriosis in a hooded capuchin (Sapajus apella).

Spontaneous endometriosis is common in women, great apes and Old World monkeys, but rare in New World monkeys. We report on the clinical diagnosis and successful combination of surgical and medical treatment in a case of spontaneous endometriosis in a diabetic hooded capuchin. Addressing this painful condition had a positive welfare impact.