Kibble diet is associated with higher faecal glucocorticoid metabolite concentrations in zoo-managed red wolves (Canis rufus).

The red wolf (Canis rufus) is a critically endangered canid that exists solely because of the establishment of the ex situ population in the late 1980s. Yet, the population under human care suffers from gastrointestinal (GI) disease in captivity. While the cause of GI disease is unknown, it is speculated that environmental factors can influence GI health of zoo-managed red wolves. The goal of the present study was to investigate the relationship between faecal glucocorticoid metabolite (FGM) concentrations, a biomarker for stress, and environmental factors for zoo-managed red wolves. Faecal samples were collected from 14 adult wolves three times a week for 5 to 12 months. Using a single-antibody cortisol enzyme immunoassay, FGM concentrations were quantified. Environmental factors were collected for each participating wolf on dietary type, sex, type of public access to enclosure, density (enclosure size [ft2]/number of wolves living in enclosure) and a monthly average status of GI health. Red wolves that ate a commercial kibble diet had both higher FGM concentrations over time and higher baseline FGM concentrations compared to individuals that received commercial kibble mixed with commercial meat. Density, public access or GI health were not related to FGM concentration; however, males had higher baseline FGM concentrations compared to female red wolves. Our findings suggest that management conditions, particularly diet, can strongly influence FGM concentration in the zoo-managed red wolf population. Findings from this study highlight the importance of management choices on individual welfare. Maintaining a healthy captive population of red wolves is imperative for the persistence of the species, including successful future reintroductions.

Hormonal variation and temporal dynamics of musth in Asian elephants (Elephas maximus) are associated with age, body condition and the social environment.

The sustainability of endangered Asian elephants in human care is threatened in part by low breeding success and concerns over individual animal wellbeing. Male elephants have received less research attention compared to females, yet males deserve special consideration due to their unique reproductive biology (particularly the sexual state of "musth") and the complex interaction of physiological, environmental, and social pressures they face. We measured fecal androgen metabolites (FAMs), fecal glucocorticoid metabolites (FGMs), and fecal triiodothyronine metabolites (FT3s) collected weekly over approximately 12 months from 26 male Asian elephants housed in zoos across the US, hypothesizing that FAM, FGM, and FT3 concentrations would be associated with temporal correlates of musth and would vary further with intrinsic (musth status, age, body condition) and extrinsic (social environment) factors. The duration of each musth episode was positively associated with exposure to male conspecifics and negatively associated with body condition. Further, elevated FAM concentrations were associated with social exposure, age, and body condition, and FGM concentrations also varied with age and body condition. FT3 concentrations were not associated with any factor we measured. We also identified periods of lower FAM concentration than confirmed musth episodes (but still higher than baseline FAM concentrations) that we termed "elevated FAM episodes." The durations of these episodes were negatively correlated with exposure to other male elephants. Together, these results provide evidence that hormone profiles (including those that are predicted to change around musth) vary significantly between male Asian elephants in a way that may be attributed to intrinsic and extrinsic factors. Studies like these serve to enhance the sustainability of ex-situ populations by providing wildlife managers with information to enhance the health, welfare, and reproduction of threatened species like Asian elephants.

Social Behavior and Group Formation in Male Asian Elephants (Elephas maximus): The Effects of Age and Musth in Wild and Zoo-Housed Animals.

Asian elephants are endangered, and the long-term viability of the species depends on integrative approaches to address the sustainability of in-situ and ex-situ populations. Growing evidence shows that male elephants exhibit extensive and flexible social behavior that rivals the complexity of that of females. Male elephant sociality is expected to change dramatically around the unique sexual state of musth. However, data related to male Asian elephant sociality is lacking. Here, we conducted complementary observations in Wasgamuwa National Park, Sri Lanka, and North American zoos of male Asian elephant social behavior. Age and musth status, along with other factors, were associated with variation in social behavior and group formation of males. In wild male elephants, both musth status and age impacted elephant associations within all-male and mixed-sex groups: non-musth elephants were generally sighted less often in mixed-sex groups as they aged, while the inverse occurred with musth elephants. Musth status interacted with age to predict the number of conspecifics with which a wild male elephant associated: younger males were observed with more females during non-musth (but the opposite was true during musth), and male elephants between 20 and 30 years were observed with the highest number of male conspecifics except during musth. Finally, we found variation in aggression, prosocial behavior, and submissive behavior was influenced by intrinsic (age and musth status) and extrinsic factors (group size and type) in similar ways in both populations; prosocial behavior was most common and was influenced by the number of conspecifics present (both populations), and age, group type, and musth status (zoo population), while aggression was rare, especially among older elephants. We suggest that longitudinal studies of this threatened species will be particularly helpful to promote the reproduction and conservation of Asian elephants in in-situ and ex-situ environments.

Influence of Reproductive Status on Equine Serum Proteome: Preliminary Results.

The reproductive cycle and early pregnancy represent dynamic physiological states in mammals, but mechanisms involved in early pregnancy in the domestic horse remain poorly understood. Proteins in uterine secretions have been studied, but the proteome of peripheral serum during various reproductive states has not been investigated. This study characterized and compared the serum proteome in the domestic horse during various reproductive states. Serum was collected from three mares during: (1) estrus (day [d] -1; d 0 = ovulation), (2) diestrus (d 12.5, non-mated), (3) early pregnancy (d 12.5, pregnant), and (4) nonpregnant (d 12.5, unsuccessfully mated) states. Serum proteins in each sample were analyzed by Nano LC-MS/MS, and 308 proteins were identified. Differentially-expressed proteins (DEP; > 1.5-fold or < - 0.5-fold) were identified by comparison of protein relative abundance between reproductive states: (1) diestrus compared to estrus (DEP = 71), (2) pregnant compared to diestrus (DEP = 72), and (3) non-pregnant compared to pregnant (DEP = 81). DEPs were analyzed for biological function using PANTHER (pantherdb.org). Several pregnancy-specific proteins previously identified in equine pregnant histotroph, including Apolipoprotein A-I, Complement C3, and Histone H4, were detectable in the serum. The ability to detect these biomarkers in serum provides a more readily available option for investigating and understanding early equine pregnancy.

Differential expression of urinary volatile organic compounds by sex, male reproductive status, and pairing status in the maned wolf (Chrysocyon brachyurus).

The maned wolf (Chrysocyon brachyurus) is an induced ovulator. Though the mechanism of ovulation induction remains unknown, it is suspected to be urinary chemical signals excreted by males. This study assessed volatile organic compounds (VOCs) in weekly urine samples across 5 months from 13 maned wolves (6 intact males, 1 neutered male, 6 females) with the goal of identifying VOCs that are differentially expressed across sex, reproductive status, and pairing status. Solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) were used to extract and separate VOCs that were identified via spectral matching with authentic standards, with spectral libraries, or with new software that further matches molecular fragment structures with mass spectral peaks. Two VOCs were present across all 317 urine samples: 2,5-dimethyl pyrazine and 2-methyl-6-(1-propenyl)-pyrazine. Fifteen VOCs differed significantly (Adj. P < 0.001 and |log2 fold change| >2.0) between intact males and females. Using partial least squares-discriminant analysis, the compounds with the highest importance to the sex classification were delta-decalactone, delta-dodecalactone, and bis(prenyl) sulfide. Sixty-two VOCs differed between intact males and the neutered male. Important classifier compounds were 3-ethyl 2,5-dimethyl pyrazine, 2-methyl-6-(1-propenyl)-pyrazine, and tetrahydro-2-isopentyl-5-propyl furan. Several VOCs established as important here have been implicated in reproductive communication in other mammals. This study is the most robust examination of differential expression in the maned wolf thus far and provides the most comprehensive analysis of maned wolf urinary VOCs to date, increasing the sample size substantially over previous chemical communication studies in this species. New data analysis software allowed for the identification of compounds in the hormone-producing mevalonate pathway which were previously unreported in maned wolf urine. Several putative semiochemicals were identified as good candidates for behavioral bioassays to determine their role in maned wolf reproduction, and specifically in ovulation induction.

Gut Microbiomes Differ Among Dietary Types and Stool Consistency in the Captive Red Wolf (Canis rufus).

Captive management of many wildlife species can be challenging, with individuals displaying health disorders that are not generally described in the wild population. Retrospective studies have identified gastrointestinal (GI) diseases, in particular inflammatory bowel disease (IBD), as the second leading cause of captive adult red wolf (Canis rufus) mortality. Recent molecular studies show that imbalanced gut microbial composition is tightly linked to IBD in the domestic dog. The goal of the present study was to address two main questions: (1) how do red wolf gut microbiomes differ between animals with loose stool consistency, indicative of GI issues, and those with normal stool consistency and (2) how does dietary type relate to stool consistency and red wolf gut microbiomes? Fresh fecal samples were collected from 48 captive wolves housed in eight facilities in the United States and from two wild wolves living in Alligator River National Wildlife Refuge, NC, United States. For each individual, the stool consistency was categorized as loose or normal using a standardized protocol and their diet was categorized as either wild, whole meat, a mix of whole meat and kibble or kibble. We characterized gut microbiome structure using 16S rRNA gene amplicon sequencing. We found that red wolves with a loose stool consistency differed in composition than wolves with normal stool consistency, suggesting a link between GI health and microbiome composition. Diet was not related to stool consistency but did significantly impact gut microbiome composition; gut microbiome composition of wolves fed a kibble diet were significantly different than the gut microbiome composition of wolves fed a mixed, whole meat and wild diet. Findings from this study increase the understanding of the interplay between diet and GI health in the red wolf, a critical piece of information needed to maintain a healthy red wolf population ex situ.

Utilizing Camera Traps, Closed Circuit Cameras and Behavior Observation Software to Monitor Activity Budgets, Habitat Use, and Social Interactions of Zoo-Housed Asian Elephants (Elephus maximus).

Accredited zoos and aquariums value superior animal husbandry and strive to ensure that the physical, psychological, and social needs of animals are met. In North America, the Association of Zoos and Aquariums (AZA) relies on species-specific standards to ensure facilities provide the best care for collection animals. The AZA also makes explicit recommendations for long-term monitoring of welfare. Data collected through behavioral observations can be used to modify management as animals respond over time to social, environmental, or physical changes. In long-lived, social species like elephants, it is particularly important to document herd dynamics, calf development, geriatric health, and social bonds throughout their lifetimes. The Smithsonian's National Zoological Park housed one male and six female Asian elephants in dynamic social groupings. Behavioral observations were conducted on all elephants for two years using two methods involving ZooMonitor, closed circuit cameras, and camera traps. The goal was to compare how these two methods function to provide individual activity budgets, habitat use, and social interactions. Methodologies such as these, alone or in combination, have the potential to produce valuable data about potential changes in welfare over time in a zoological setting and can be performed either by staff or volunteers with high reliability.

Influence of Metabolic Status and Diet on Early Pregnant Equine Histotroph Proteome: Preliminary Findings.

Insulin resistance (IR) is characterized by an increase in biomarkers of systemic inflammation and susceptibility to laminitis in horses. Impacts on reproduction include a lengthened interovulatory period in horses. Dietary omega-3 (docosahexaenoic acid [DHA]) promotes anti-inflammatory processes, has been implicated in health benefits, and can reduce cytokine secretion. This preliminary study investigated the impact of IR as well as the influence of dietary supplementation (DHA) on the uterine fluid proteome in early pregnant horses. Mares were artificially inseminated; uterine fluid and embryos were collected on d 12.5 after ovulation. Uterine fluid was pooled for metabolic and diet categories (n = 8; n = 2 per metabolic and dietary status) and concentrated, and the proteome was analyzed using tandem mass spectrometry (iTRAQ). Five proteins met differential abundance criteria (±1.5-fold change, P < .05) in all comparisons (Control C, IS vs. C, IR; C, IS vs. DHA, IS; C, IR vs. DHA, IR). Serum amyloid A, afamin, and serotransferrin were upregulated in C, IR mares but downregulated in DHA, IR mares when compared to C, IS and C, IR, respectively. Quantitative PCR supported mass spectrometry results. The presence of serum amyloid A and serotransferrin in histotroph of IR mares potentially indicates an inflammatory response not seen in IS counterparts. These preliminary findings provide novel evidence on the potential impact of insulin resistance and DHA supplementation on the secreted equine uterine proteome during early pregnancy.

Longitudinal fecal hormone monitoring of adrenocortical function in zoo housed fishing cats (Prionailurus viverrinus) during institutional transfers and breeding introductions.

The ex situ population of fishing cats (Prionailurus viverrinus) has become increasingly important for the conservation of this species. Unfortunately, captivity-induced stress is a concern and potential factor for lack of breeding success in this small felid, resulting in an unsustainable population. The objectives of this study were to: 1) validate an enzyme immunoassay for monitoring of fecal glucocorticoid metabolite (FGM) concentrations in the fishing cat; 2) identify potential exogenous stressors in the captive environment; 3) pinpoint management techniques that may lower FGM concentrations; and 4) determine if FGM concentrations are related to breeding success. Through a successful adrenocorticotrophic hormone challenge and additional laboratory methods, a cortisol enzyme immunoassay was validated as an effective tool for detecting FGM in this species. Between 2010 and 2013, longitudinal FGM monitoring was conducted in 26 fishing cats in the North American Species Survival Plan®. Exogenous stressors that elevated FGM concentrations included: chemical immobilizations; permanent transfers between facilities; construction; facility events; and fights/aggression among breeding pairs. Management factors that lowered FGM concentrations included: increased animal-keeper interaction through formal training; and providing indoor, off-exhibit, holding areas. In addition, social housing of individuals (either established breeding pairs or same sex pairs) decreased FGM concentrations. Individuals with breeding success (defined as observed copulations during the study period) also had lower FGM concentrations than unsuccessful individuals. Findings indicate that management factors play a role in lowering glucocorticoid (stress) levels in fishing cats, which may ultimately affect breeding success in the ex situ population.

Evaluation of management in North American zoos to enhance breeding success of the fishing cat (Prionailurus viverrinus) ex situ population.

The ex situ population of fishing cats (Prionailurus viverrinus) face many challenges to its sustainability such as mate incompatibility, low founder numbers and disease prevalence. The North American population was monitored for a three-year period during institutional transfers and breeding introductions. In total, 26 fishing cats, including 15 different breeding pairs were monitored during 20 transfers. Most institutional transfers occurred in the fall months (September, October and November; 62%; n = 13) and males were transferred more often (62%; n = 13). Breeding success (observed copulations) was recorded in 33% (n = 5) of pairs but only 13% (n = 2 pairs) produced offspring during the study period. Institutions with successful breeding pairs had a greater number of indoor, off-exhibit enclosures (2.67 ± 0.29 enclosures; n = 5) compared to facilities with unsuccessful pairs (1.69 ± 0.25 exhibits; n = 6; p = 0.035). In addition, facilities housing successful pairs performed positive reinforcement training more frequently (14.77 ± 3.27 training days/month) than facilities with unsuccessful pairs (4.00 ± 2.73 days/month; p = 0.035). A binomial generalized linear model showed that friendly vocalizations (p = 0.000) during physical introductions of intended breeding pairs predicted copulation success. Introductions performed when a female was exhibiting estrous behavior (p = 0.020), was also predictive of copulation success. Results from this study are the first comprehensive analyses of captive management in the fishing cat. Environmental factors and management approaches are highlighted that could advance animal welfare and improve reproductive success in this species.

Physiological impacts of housing maned wolves (Chrysocyon brachyurus) with female relatives or unrelated males.

The maned wolf is a threatened canid species native to South America. Previous studies have suggested the species exhibits induced ovulation. In captive breeding facilities, reproductive success is low while rates of neonatal mortality are high. Females that are not recommended for breeding are frequently housed together. However there has never been a systematic study of the reproductive consequences of co-housing females. This study was conducted for three purposes, to: (1) corroborate the presence of induced ovulation, (2) determine whether elevated cortisol is implicated in neonatal pup mortality, and (3) evaluate the endocrine correlates of group housed females. Using fecal hormone monitoring for estrogen, progesterone, and cortisol, 43 cycles from 33 female maned wolves were studied from 2002 to 2015. Females were categorized by their reproductive status: pregnant and successfully raised pups (PR; n = 11), pregnant with neonatal pup demise within 3 days (PL; n = 7), housed with a male but no signs of breeding or pregnancy (PP; n = 10), housed singly (S; n = 8), or housed with related females (F; n = 7). Estrogen and progestagen remained at baseline for all females not housed with a male (S, F), while elevations consistent with ovulation were seen in females housed with a male (PP, PL, PR). Compared to PR females, PL individuals showed similar cortisol levels throughout the cycle and slightly lower progesterone levels during gestation. As for the effect of co-housing related females, F females showed estrogen and progesterone levels lower even than S females while cortisol levels were elevated compared to all other groups. These findings support the previous evidence of induced ovulation in the maned wolf. Although elevated cortisol does not seem to be implicated in pup loss, a non-significant trend towards lower progesterone during gestation could be implicated. Future studies should assess depressed progesterone levels as a correlate to neonatal pup mortality. Female maned wolves housed with related females experience suppressed reproductive hormones and elevated adrenal hormones. Therefore, a more systematic study of hormonal and behavioral correlates to co-housing with related females is warranted.

Group management influences reproductive function of the male cheetah (Acinonyx jubatus).

Although the free-ranging cheetah is generally socially solitary, as many as 60% of males live in same-sex (usually sibling) coalitions. Under ex situ conditions, the cheetah experiences low reproductive success with only ~18% of males having ever produced young. Most male cheetahs (85%) are managed in captivity in coalitions, but with no data on the influence of social grouping on reproductive parameters. We examined the influence of singleton versus coalition management on various male cheetah physiological traits, including ejaculate quality and gonadal and adrenal hormone metabolite concentrations. We also assessed behaviour within coalitions for evidence of social hierarchy through initiation of interactions with group mates and relatedness to physiological traits. Ejaculate quality (including total motile and structurally normal spermatozoa per ejaculate) and androgen concentration profiles were higher (P<0.05) in coalition compared with singleton males. These results support the conclusion that testis function in the cheetah, specifically related to the development of normal, motile spermatozoa and androgen production, is influenced by management with same-sex conspecifics. The findings have implications for ex situ conservation breeding programs by suggesting that reproductive quality can be enhanced through group maintenance of cheetah males.

Motile Sperm Output by Male Cheetahs (Acinonyx jubatus) Managed Ex Situ Is Influenced by Public Exposure and Number of Care-Givers.

The collective cheetah (Acinonyx jubatus) population in zoological institutions has never been self-sustaining because of challenges in natural reproduction. A retrospective analysis of North American zoo-breeding records has revealed that >90% of litters produced since 2003 occurred in facilities 'off-display' from the public. We examined seminal, endocrine, and behavioral traits of 29 adult male cheetahs that were: 1) managed in public exhibit or off-display facilities; 2) maintained by different numbers of cheetah-specific care-givers; and 3) living adjacent to varying numbers of adult conspecifics. Cheetahs housed off-display produced more total motile sperm/ejaculate (P = 0.04) than on-exhibit males. This finding was mirrored in our laboratory's historical records where two-fold more total motile sperm (P < 0.01) were measured in ejaculates from individuals with no public exposure (n = 43) compared to on-exhibit (n = 116) counterparts. Males at institutions with ≤3 care-givers also produced more total motile sperm/ejaculate (P < 0.03) and spent more time behaviorally active (P < 0.01) than at facilities using >3 care-givers. Exposure to high numbers of conspecifics within the same institution did not impact (P > 0.05) seminal traits, and presence of the public, care-giver number, or animals/facility had no influence (P > 0.05) on androgen or glucocorticoid excretion or other behavioral metrics. Findings indicate that male cheetahs are sensitive to general public exposure and too many care-givers, resulting in compromised motile sperm output/ejaculate with mechanism of action unrelated to altered androgen or glucocorticoid excretion.

Induction of ovarian activity and ovulation in an induced ovulator, the maned wolf (Chrysocyon brachyurus), using GnRH agonist and recombinant LH.

Assisted reproductive techniques, such as ovarian manipulation and artificial insemination, are useful for enhancing genetic management of threatened wildlife maintained ex situ. In this study, we used noninvasive fecal hormone monitoring to investigate (1) the influence of pairing with a male on endocrine responses of female maned wolves (Chrysocyon brachyurus) to a GnRH agonist (deslorelin) and (2) the efficiency of recombinant LH (reLH) on ovulation induction in females housed alone. Deslorelin (2.1 mg Ovuplant) was given to females that were either paired with a male (n = 4) or housed alone (n = 7); the implant was removed 7 to 11 days postimplantation. Three of seven singleton females were injected with reLH (0.0375 mg) on the day of implant removal, whereas the remaining females (n = 4) did not receive the additional treatment. Fecal samples were collected 5 to 7 days/wk from all females starting 11 days prior to hormone insertion until at least 70 days post implant removal for a total of 11 hormone treatment cycles. Fecal estrogen and progestagen metabolites were extracted and analyzed by enzyme immunoassay. Evidence of ovulation, demonstrated by a surge of estrogen followed by a significant rise in progestagen, occurred in all paired females. Three of the four singleton females that did not receive reLH treatment exhibited no rise in progestagen after an estrogen surge. All singleton females treated with reLH exhibited a rise in fecal progestagen after injection, indicating ovulation. In conclusion, deslorelin is effective at inducing ovarian activity and ovulation in paired female maned wolves; however, exogenous reLH is needed to induce ovulation in females housed alone. The findings obtained from this study serve as a foundation for future application of artificial insemination to enhance genetic management of this threatened species ex situ.

Spermatozoa from the maned wolf (Chrysocyon brachyurus) display typical canid hyper-sensitivity to osmotic and freezing-induced injury, but respond favorably to dimethyl sulfoxide.

We assessed the influences of medium osmolality, cryoprotectant and cooling and warming rate on maned wolf (Chrysocyon brachyurus) spermatozoa. Ejaculates were exposed to Ham's F10 medium (isotonic control) or to this medium plus NaCl (350-1000mOsm), sucrose (369 and 479mOsm), 1M glycerol (1086mOsm) or dimethyl sulfoxide (Me2SO, 1151mOsm) for 10 min. Each sample then was diluted back into Ham's medium and assessed for sperm motility and plasma membrane integrity. Although glycerol and Me2SO had no influence (P>0.05), NaCl and sucrose solutions affected sperm motility (P<0.05), but not membrane integrity. Motility of sperm exposed to <600mOsm NaCl or sucrose was less (P<0.05) than fresh ejaculate, but comparable (P>0.05) to the control. As osmolality of the NaCl solution increased, motility decreased to <5%. In a separate study, ejaculates were diluted in Test Yolk Buffer containing 1M glycerol or Me2SO and cooled from 5°C to -120°C at -57.8°C, -124.2°C or -67.0°C/min, frozen in LN2, thawed in a water bath for 30s at 37°C or 10s at 50°C, and then assessed for motility, plasma- and acrosomal membrane integrity. Cryopreservation markedly (P<0.05) reduced sperm motility by 70% compared to fresh samples. Higher (P<0.05) post-thaw motility (20.0±1.9% versus 13.5±2.1%) and membrane integrity (51.2±1.7% versus 41.5±2.2%) were observed in samples cryopreserved in Me2SO than in glycerol. Cooling rates influenced survival of sperm cryopreserved in glycerol with -57.8°C/min being advantageous (P<0.05). The findings demonstrate that although maned wolf spermatozoa are similar to domestic dog sperm in their sensitivity to osmotic-induced motility damage, the plasma membranes tolerate dehydration, and the cells respond favorably to Me2SO as a cryoprotectant.

Effects of management strategies on glucocorticoids and behavior in Indian rhinoceros (Rhinoceros unicornis): translocation and operant conditioning.

The ex situ Indian rhino population experienced a decrease in genetic diversity indicating that the breeding program could possibly benefit from novel reproductive management strategies to ensure population sustainability. We sought to determine how management tools used for reproductive management, specifically translocation and operant conditioning, impact physiological and behavioral measures of welfare in Indian rhinos. First, an adrenocorticotropic hormone challenge performed in an adult male resulted in a 38-fold increase in urinary and a 3.5-fold increase in fecal glucocorticoid metabolites (FGM). Mean and peak FGM differed among three females, but all demonstrated elevated (P < 0.0001) concentrations for variable durations after translocation that lasted up to 9 weeks. Lastly, behavioral and adrenal responses of two females to operant conditioning to stand during transrectal ultrasound exams were monitored and rhinos differed in their mean and peak FGM concentrations. However, FGM were not different before versus during training or on pasture versus in the barn. One female exhibited more stereotypic behavior during training in the barn than on pasture (P < 0.05); although, stereotypies (1.73% of time) were relatively uncommon overall. In summary, individual variation exists in FGM both at baseline levels and in response to a stressor. In addition, while a transient rise in glucocorticoid activity post-translocation indicated that Indian rhinos have a physiological response to changes in their environment, minor alterations in daily routines using operant conditioning only resulted in minimal changes in behaviors and FGM.

Impacts of environmental pressures on the reproductive physiology of subpopulations of black rhinoceros (Diceros bicornis bicornis) in Addo Elephant National Park, South Africa.

Black rhinoceros are an icon for international conservation, yet little is known about their physiology due to their secretive nature. To overcome these challenges, non-invasive methods were used to monitor rhinoceros in two sections of Addo Elephant National Park, South Africa, namely Addo and Nyathi. These sections were separated by a public road, and the numbers of elephants, predators and tourists were higher in Addo. Faecal samples (n = 231) were collected (from July 2007 to November 2010) from known individuals and analysed for progestagen and androgen metabolite (FPM and FAM, respectively) concentrations. As biotic factors could impact reproduction, we predicted that demographics, FPM and FAM would vary between sections and with respect to season (calendar and wet/dry), climate and age of the rhinoceros. Mean FPM concentrations from pregnant females were seven times higher (P < 0.05) than samples from non-pregnant rhinoceros. Positive relationships were found between monthly temperatures and FPM from non-pregnant females (r (2) = 0.25, P = 0.03) and the percentage of calves born (r = 0.609, P = 0.04). Although FAM peaked in the spring, when the majority of calves (40%) were conceived, no seasonal patterns in male androgen concentrations were found with respect to month of conception and parturition. Females in Addo had a longer inter-calving interval and were less likely to be pregnant (P < 0.05) compared with those in Nyathi. The biotic stressors (e.g. predators and more competitors) within Addo section could be affecting the reproductive physiology of the rhinoceros negatively. Enhanced knowledge about how black rhinoceros populations respond to environmental stressors could guide management strategies for improving reproduction.

Ovarian cycle activity varies with respect to age and social status in free-ranging elephants in Addo Elephant National Park, South Africa.

Free-ranging African elephants live in a fission-fusion society, at the centre of which is the matriarch. Matriarchs are generally older females that guide their families to resources and co-ordinate group defense. While much is known about elephant society, knowledge is generally lacking about how age affects the physiology of wild elephants. Investigation of the ovarian activity of free-ranging elephants could provide insight into the reproductive ageing process, with implications for population management. Faecal samples were collected from 46 individuals ranging in age from 14 to 60 years for a 2-year period, and progestagen metabolite analyses were used to examine relationships between social status, age, season, and ovarian activity in female elephants in Addo Elephant National Park, South Africa. Social status was the strongest predictor of faecal progestagen metabolite concentrations in non-pregnant elephants, with grand matriarchs (n = 6) having the lowest values compared with matriarchs (n = 21) and non-matriarch females (n = 19). Likewise, social status and age were the strongest predictors of faecal progestagen metabolite concentrations in pregnant elephants (n = 27). The number of years since a non-pregnant female gave birth to her last calf (post-partum duration) was longer for older females with a higher social status, as well as during the dry season. Our results indicate that social standing and age of elephants are related to reproductive function, and that older females exhibit reductions in ovarian capacity. These results expand our understanding of reproduction and fertility throughout an elephant's lifespan, and the factors that impact gonadal function in free-ranging females. Given that possible over-abundance of elephants in areas such as Addo Elephant National Park is fuelling the debate over how best to manage these populations, knowledge about the reproductive potential of high-ranking females can provide managers with biological data to identify the best candidates for controlling growth through translocation or contraception.

Characterizing sleep behavior of the wild black rhinoceros (Diceros bicornis bicornis).

STUDY OBJECTIVES: The objectives of this study were to characterize sleep patterns and determine factors, including sex, age, season, and environmental pressures, that influence sleep in the endangered black rhinoceros (rhino; Diceros bicornis bicornis). DESIGN: To noninvasively observe sleep behavior of wild rhinos, digital infrared cameras were erected on poles at two bedding sites from September 2009 to March 2010. SETTING: The study site was located in South Africa's Addo Elephant National Park (AENP) in the Main Camp (Addo) and Nyathi sections. PARTICIPANTS: A total of 2,417 photos captured rhino sleep behavior on eight individual rhinos during 40 separate sleeping bouts (Addo, n = 15; Nyathi, n = 25). INVENTIONS: N/A. RESULTS: Results demonstrated that age and season did not affect rhino sleep behavior (P > 0.05); however, sex did influence the length of sleep bouts with males (n = 27; mean, 105.6 ± 11.3 min; range, 14.0-202.0 min) sleeping longer (F(1,48) = 6.93, P = 0.01) than females (n = 13; mean, 58.6 ± 10.4 min; range, 11.0-132.0 min). Park section did not influence the length of sleep episodes, but did affect (rw(40) = 0.88; P < 0.025) the time at which rhinos slept (Addo, 20:00-24:00; Nyathi, 20:00-04:00). CONCLUSIONS: This is the first study to characterize sleep behavior in wild black rhinos. This study resulted in a greater understanding of the biologic factors that affect sleep in wild rhinos and can provide information to assist their management and conservation.

International Society for Wildlife Endocrinology: the future of endocrine measures for reproductive science, animal welfare and conservation biology.

Hormone analysis is a precise and widely accepted tool for monitoring reproductive function and responses to stressors. Although hormones are present and can be measured in various biological matrices, non-invasive methods have gained popularity over the past 30 years as a more practical approach for assessing ovarian, testicular and, more recently, adrenocortical activity in intractable wildlife species. Non-invasive hormone monitoring also has been key to understanding biological mechanisms related to observed behaviours of captive and free-ranging animals. Despite the increasing popularity of this research field, wildlife endocrinologists have not had a specific forum for sharing and discussing their latest findings, technical developments and common challenges. To provide such a communication platform, the International Society for Wildlife Endocrinology (ISWE) was established in 2010, followed by an international meeting held on 3-4 November 2011 at the Toronto Zoo, Canada. Over several sessions, keynote speakers and participants discussed recent developments of new and innovative methods for hormone monitoring, as well as the latest advances in basic endocrinology as applied to adrenal function, reproductive physiology, animal health, ecology and evolution. Here, we introduce ISWE to the scientific community and discuss how this new society will serve as a resource for wildlife endocrinologists worldwide.