Ex-situ conservation strategies such as the formation of somatic cell banks are valuable tools for the conservation of jaguars, whose population has been declining in recent years. Once properly established, these cells can be successfully leveraged for future applications. We aimed to assess the effects of in vitro culture and cryopreservation on the establishment of fibroblasts derived from jaguars. Initially, we identified five dermal fibroblastic lines using morphology and immunophenotyping assays; these lines were then subjected to two experiments. In the first experiment, the viability, metabolism, and proliferative activity of cells at different passages (first, third, and tenth) were evaluated. In the second experiment, the cells were cryopreserved and the levels of reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm) and apoptosis were evaluated after one, three, and ten passages. Noncryopreserved cells were used as controls. The in vitro culture after first, third, and tenth passages and cryopreservation conditions did not affect the proliferative activity and viability. However, cells cultured until tenth passage and frozen/thawed cells showed reduced metabolism. In addition, cryopreserved cells showed higher levels of intracellular ROS and altered ΔΨm when compared with those of noncryopreserved cells. Finally, frozen/thawed cells cultured after ten passages showed reduced proliferative activity and number of viable cells than did frozen/thawed cells cultured after one and three passages. In summary, we have shown that viable fibroblasts can be established from jaguar skin and that although these cells do not show altered viability and proliferative activity, they do undergo damage during extended culture and cryopreservation.

The giant anteater (Myrmecophaga tridactyla) is being threatened by natural habitat destruction and fragmentation, illegal hunting and road kills. In this context, the generation of basic information on the reproductive parameters of this species is vital, aiming to improve reproductive management via, amongst others, assisted reproductive technologies. This study aimed to describe the morphological and functional features of semen collected from captive giant anteaters. Electroejaculation was performed in 13 animals housed in zoos located in São Paulo state, Brazil. Semen samples were collected from 13 animals in 16 procedures. Samples were evaluated for volume, motility, vigor, pH, concentration, sperm morphology, and functional tests. The following mean values were obtained: volume 1.28 ± 0.27 mL; motility 28.3 ± 6.2%; vigor 2.4 ± 0.25; concentration 129.4 ± 36.1 × 106 sperm/mL; pH 7.4 ± 0.2. Total acrosome, head, midpiece, and tail sperm abnormalities were 3.2 ± 0.8%, 25.4 ± 3.6%, 20.7 ± 3.2%, and 14.7 ± 2.6%, respectively. Intact acrosome was found in 83.7 ± 3.1% and intact membrane in 81.1 ± 4.0% of all samples collected. Mitochondrial activity was 66.4 ± 6.0% (Class I), 18.7 ± 2.9% (Class II), 8.0 ± 2.0% (Class III), 3.9 ± 1.0% (Class IV), and 3.0 ± 0.9% (Class V). Sperm DNA fragmentation rate was 13.2 ± 3.7%. These results indicated that electroejaculation is a feasible method for semen collection in giant anteaters, allowing a more detailed description of the semen in this species.

The ploughshare tortoise (Astrochelys yniphora) is an endangered species. It is important to increase the number of tortoises in the wild and captivity; however, breeding in captivity, especially outside of the natural habitat area, has been unsuccessful worldwide. As a result of an emergency rescue due to a violation of the Act of Conservation of Endangered Species of Wild Fauna and Flora, Nogeyama Zoological Gardens(Yokohama, Japan) has housed ploughshare tortoises, and a natural breeding trial started in 2011. In this case, the captive environment in Nogeyama, especially room temperature and humidity, imitated the environment in the natural habitat of northwest Madagascar. The maximum temperature during the day was 30.8-32.5°C throughout the year, but the minimum temperature was 24°C during December and January, representing the rainy season, and 17°C during July, representing the dry season. The humidity was adjusted and the room was not humidified between November and February (dry season)but was humidified by sprinkling water once every few days in March and daily between April and October (rainy season). Natural breeding and egg-laying were observed 1 year after the captive environmental change. Three neonates were hatched from nine eggs in 3 years after the beginning of the breeding program. Thisstudy showed that for natural breeding of the ploughshare tortoises, it is important to adjust the temperature and humidity of the captive environment to imitate the temperature and humidity of the natural habitat.

A decline in sperm quality with age is a common prediction of senescence-based hypotheses and empirical studies. While widely studied across taxa, there is little known on the effect of ageing on sperm quality in amphibians, especially in captive populations used for controlled propagation and reintroduction efforts. Here, we investigated variation in sperm quality metrics (i.e., motility, concentration, and morphology) in the endangered Mississippi gopher frog (Lithobates sevosus) among males of three age categories using individuals from captive breeding populations housed at three different zoological institutions. Different aged males across the species expectant lifespan (1-9, 1-2, 3-4, and 8-9-year-old subcategories) were chosen in an attempt to identify an optimal breeding age relevant for captive breeding programs. Moreover, we explored and statistically controlled for potential differences in sperm quality which may be attributed to the type of induction hormones and source populations that differed among institutions. Results indicated that males of different ages did not differ in sperm motility or concentration. However, we did find that older males (8-9 years old) had significantly longer sperm than other age categories and younger males (1-2 years old) had significantly more atypical sperm than other age categories. Furthermore, we found no significant differences in any sperm quality metrics between the different induction hormones or source populations used at the different institutions. Within a captive breeding program, this information is especially valuable as our results indicate that males that have only recently sexually matured may not be ready to breed, while older males maintain sperm quality metrics presumably related to fertilization success.

Birth events across sixteen different mammalian species representing four orders were randomly dispersed through the week with no evidence that any species avoided giving birth when the zoos were most crowded with visitors indicating visitors were not sufficiently stressful to bring about birth delays.

The impact of visitors on the well-being of captive animals presents both positive and potentially negative consequences. While some amount of novelty through visitor stimulation offers the opportunity for a more complex captive environment, anecdotal evidence from primate staff observations at the Smithsonian's National Zoo and Conservation Biology Institute suggested that gorillas exhibited increased restlessness during the annual month-long late night ZooLights (ZL) event than before it. The current study compared activity budgets, aggression (interactions involving contact between conspecifics and displays toward visitors), and abnormal behaviors in two groups of socially housed gorillas for 1-month periods before, during, and after the 2015 ZL event. We also compared the fecal glucocorticoid metabolite profiles of all six gorillas across these same observation periods. Physiologically, most individuals appeared to cope appropriately with the increased visitor presence during the event. We saw little difference in contact aggression; however, abnormal behavior was observed in some gorillas during and after the event, which highlights the importance of individual analysis and data interpretation. As predicted, we found that the majority of gorillas rested less during ZL than during other observation periods, particularly adult females in the mixed sex troop. Preliminary results of this study aided the decision of zoo management to close initially the Great Ape House and subsequently most animal buildings during future ZL events to avoid the potential disruption of normal activity patterns. While the findings of this study were mixed as to the impact of the event on these gorillas, the results suggest that zoos should carefully consider the possible ramifications of keeping great ape buildings open for lengthy multi-night events.

The Puerto Rican Boa (Chilabothrus inornatus) was placed on the US Endangered Species List in 1970. Progress has been made since to clarify the recovery status of this species, though the design of a new recovery plan must include information regarding genetic variation within and among populations of this species. While measures of genetic diversity in wild populations of this species are finally becoming available, relative genetic diversity represented in ex situ populations is unknown, which hampers efforts to develop an ex situ species management plan. Here, we provide an analysis of genetic diversity in US public and private collections (zoos and breeders) using mitochondrial sequence data and five highly polymorphic nuclear microsatellite loci. We analyzed 50 boas from the US ex situ population and determined overall genetic diversity and relatedness among these individuals. We then compared these data to mitochondrial and microsatellite data obtained from 176 individuals from wild populations across the native range of the species. We found little inbreeding and a large amount of retained genetic diversity in the US ex situ population of C. inornatus relative to wild populations. Genetic diversity in the ex situ population is similar to that found in wild populations. Ours is only the second explicit attempt to characterize genetic diversity at the molecular level in ex situ populations of boid snakes. We anticipate that these results will inform current breeding strategies as well as offer additional information that will facilitate the continuation of ex situ conservation breeding or management in boas.

Japanese macaque societies are characterized by frequent intragroup conflict both in free-ranging and zoo-housed conditions. In zoos, understanding the factors that contribute to wounding is of interest because this knowledge can aid in proactive husbandry and management planning that can minimize the negative impacts wounding can have on individual welfare. This study sought to determine whether the variables sex, season and age predicted wounding rates. Data were collected for 24 months on 119 Japanese macaques living in 10 zoos, and we analyzed the contribution of variables using generalized linear mixed models. A total of 1,007 wounds were reported, and the best model included the interaction between sex and season. Follow-up analyses revealed that females incurred more wounds than males, and this sex difference was more pronounced during the breeding compared to the nonbreeding season. On average, individuals received 4.67 (±SEM: 0.55) wounds per year and 77.31% of the population incurred at least one wound in the study period. The majority of wounds were superficial and did not require veterinary intervention. Wound locations were not randomly distributed across body regions and were most often reported on the face. Finally, macaques living in larger social groups experienced more wounds, on average. This study represents the first quantification of wounding in this species and may provide insight to help inform husbandry and management strategies in zoos.

Rest of World), 3292 (Europe), 2605 (UK