Reproductive potential and implant loss in female hamadryas baboons (Papio hamadryas) previously contracepted with melengestrol acetate contraceptive implants at AZA institutions.

Melengestrol acetate (MGA) implants are a progestin-based reversible contraceptive used to manage fertility in animals. MGA implants are recommended for replacement every 2 years; however, reproduction may be suppressed longer if implants are not removed. In this study, we investigated whether the probability of reproducing (pR) differed among nonimplanted females, females with MGA implants removed, and females whose implants were not removed. In addition, since implant loss in hamadryas baboons is a concern, we explored whether female age, institution, implant placement year, implant location, or implant placement type (intramuscular vs. subcutaneous) differed for females whose implants were lost compared to those that were not. The pR differed significantly across all three treatment conditions with the nonimplanted group having the highest pR. The pR plateaued at 63% after 40 months for the implant-removed group compared to 96% after 84 months in the nonimplanted group. There was no reproduction after contraception if implants were not removed (7.83-45.53 months). In the nonimplanted group, pR was significantly higher for older and parous females. In terms of implant loss, we found that implant placement type was significantly associated with implant loss, such that there were fewer losses when implants were placed intramuscularly (IM) as compared to subcutaneously. Our results suggest that placing MGA implants IM is likely to reduce loss. When loss is prevented, MGA implants are an effective form of contraception and are reliably reversibly in most individuals when removed. However, if not removed, they can prevent reproduction longer than 2 years.

Effects of GnRH agonists on testosterone and testosterone-stimulated parameters for contraception and aggression reduction in male lion-tailed Macaques (Macaca silenus).

Managing social groups in zoos requires controlling reproduction in individuals that do not have a current breeding recommendation, while simultaneously maintaining social harmony and animal well-being. Contraceptives, such as gonadotropin releasing-hormone (GnRH) agonists, that suppress testosterone production, offer a potential solution. They achieve infertility by interrupting spermatogenesis and may ameliorate androgen-induced aggression. This study investigated the effects of two GnRH agonists, histrelin and deslorelin, on testosterone, testis size, body weight and sperm production in male lion-tailed macaques, along with subjective observations of aggressive behavior. Five trials at three institutions with 14 males demonstrated that 100 mg histrelin or 9 to 12 mg of deslorelin could at least temporarily reduce testosterone, but a lower 6 mg dose was ineffective. However, ability of deslorelin to produce azoospermia varied among males, even at the highest dose. In general, a higher dose was needed (1) to achieve than to maintain suppression of any measured parameter and (2) to suppress sperm production than testosterone concentration. Testosterone production was also more likely than sperm production to recover, suggesting possible damage to seminiferous tubules but not to Leydig cells. Aggressive behavior was reduced in all but the group receiving the lowest dose. This allowed social groups to be maintained for many years despite recovery of testosterone in some males, suggesting that new social roles had been learned and become independent of androgen influence.

Melengestrol acetate contraceptive implant use in colobus monkeys (Colobus guereza): Patterns through time and differences in reproductive potential and live births.

Melengestrol acetate (MGA) implants are progestin-based reversible contraceptives used to manage fertility in zoo populations. Although it is recommended that MGA implants should be replaced every 2 years, the duration of efficacy has not been systematically evaluated in most species. Anecdotal reports for Old World monkeys indicate that reproduction may be suppressed longer if the implant is not removed. This study uses Guereza colobus monkey (Colobus guereza) as a model Old World monkey species to examine the effects of MGA implants on reproductive potential. In particular, we investigate whether the probability of reproducing (pR) and rates of stillbirth differ among (1) non-implanted females, (2) females who have had MGA implants removed, and (3) females whose implants were left in past expiration. We found no significant difference in pR between non-implanted and implant-removed groups, but when implants were left in past expiration, the pR was significantly lower than in other groups. Both parity and age significantly impacted pR for the non-implanted group (i.e., younger females and those who were parous increased pR), but neither were significant factors for the implant-removed group. Stillbirth rates were significantly higher post-contraception as compared with pre-contraception. These results support similar analyses in other taxa that show a shorter time to reversal after MGA contraception when implants are removed, making this a good contraceptive option for females likely to receive a breeding recommendation, especially when a more predictable time to reversal is important.

DESLORELIN (SUPRELORIN®) USE IN NORTH AMERICAN AND EUROPEAN ZOOS AND AQUARIUMS: TAXONOMIC SCOPE, DOSING, AND EFFICACY.

The Association of Zoos and Aquariums Reproductive Management Center (RMC) in the US and the European Association of Zoos and Aquaria Reproductive Management Group (RMG) in Europe monitor efficacy of contraceptive products in participating institutions and use those results to inform contraceptive recommendations. This study used the joint RMC-RMG Contraception Database to analyze efficacy of deslorelin implants (Suprelorin®), a contraceptive used in a wide range of mammalian taxa. More recently its use has increased in birds and in some reptiles and fish. Deslorelin, a gonadotropin-releasing hormone (GnRH) agonist, stimulates the reproductive system before downregulating receptors on pituitary cells that produce hormones that stimulate gonadal steroids in both males (testosterone) and females (estradiol and progesterone), interrupting sperm production and ovulation, respectively. Nevertheless, it has been used mostly in females. Efficacy has been high in mammals, with failures resulting in offspring in only 1.3% of treated individuals and 0.5% of treatment bouts. The failure rate has been higher in birds, with 14.7% of individuals in 7.2% of bouts producing eggs, perhaps reflecting differences in avian GnRH molecules. Too few reptiles and fish have been treated for meaningful analysis. Although deslorelin appears very safe, a possible exception exists in carnivores, because the stimulatory phase can result in ovulation and subsequent sustained progesterone secretion that may cause endometrial pathology. However, the stimulatory phase can be prevented by treatment with megestrol acetate for 7 d before and 7 d after implant insertion. The two current formulations of Suprelorin are effective for minimums of 6 (4.7 mg) or 12 mo (9.4 mg). The data indicate that Suprelorin is an effective and safe contraceptive option for female mammals, although it may not be effective in males of some mammalian species. Further research is needed to ascertain its usefulness in nonmammalian taxa.

Improving the sustainability of ex situ populations with mate choice.

Many breeding programs managed by the Association of Zoos and Aquariums' (AZA) Species Survival Plans® (SSPs) are not meeting goals for population size and genetic diversity due to failure of recommended pairs to breed successfully. According to AZA Population Management Center analyses, as many as 80% of recommended breeding pairs fail to produce young before the next breeding and transfer plan is issued. Determining reasons for failure and ensuring that a specific pairing produces offspring can be challenging. Mate incompatibility, one possible reason for failure, might be addressed by allowing mate choice. Although many SSP® coordinators and breeding managers, who implement breeding recommendations at their institutions, recognize the potential benefits of mate choice, examples and models for presenting and assessing choice are lacking. Here we review examples from birds, rodents, lagomorphs, marsupials, carnivores, fishes, and insects where mate choice has been incorporated. These examples provide strong evidence that free mate choice and mating with preferred partners increase a variety of reproductive success measurements when compared to assigned mate pairings. We suggest innovative housing and breeding arrangements for better incorporating mate choice into the management strategies for species held ex situ. Further, we discuss the fitness consequences and welfare implications of allowing choice. We advocate for a more systematic use of behavioral research in cooperative breeding programs. Behavioral management for mating can yield more successful programs, thus ensuring SSP® genetic and demographic goals are met, while simultaneously improving welfare.

Characterization of estrous cycles and pregnancy in Somali wild asses (Equus africanus somaliensis) through fecal hormone analyses.

Although reproduction in the domestic horse has been well described, less is known about reproduction in wild equids. This study describes endocrine patterns associated with estrous cycles and pregnancy for Somali wild asses (Equus africanus somaliensis), an endangered African equid. Fecal samples were collected three times per week for more than 2 years from five female Somali wild asses at the Saint Louis Zoo; progestagen and estrogen metabolites were quantified using commercially available immunoassays. Progestagen analysis indicated that cycle lengths were 27.2 ± 1.2 days and females cycled throughout the year. Progestagen levels during early pregnancy were low and not sustained above baseline until approximately 40 weeks prior to partition. Concentrations increased markedly around 16 weeks prior to delivery and peaked 2-3 weeks before birth. Fecal estrogen levels also increased significantly starting 40-45 weeks before parturition and reached their maximal value approximately 20 weeks prior to birth. Neither foal heat nor lactational suppression of estrus was observed, and females cycled within 45 days after delivery. These data are the first to describe the reproductive physiology of Somali wild asses. As the species faces increasing threats in the wild, this information may support conservation efforts by assisting with ex situ breeding programs.

Reactions of female cheetahs (Acinonyx jubatus) to urine volatiles from males of varying genetic distance.

Understanding the factors used by female cheetahs (Acinonyx jubatus) to make mate choice decisions could benefit zoo breeding programs, which currently assign mates based primarily on genetic distance. Because transporting animals between institutions is costly and can be stressful, females are often limited in the number of males available for mating. One solution would be to determine if an easily transported substance could be used to gauge interest by a female to a potential mate. Here, we investigate female interest in urine samples from males of different genetic distances. Twelve females at five institutions were offered scents from 17 males of varying genetic relatedness in a pair-wise choice paradigm. Behavioral responses of the females were recorded to determine preference. Results showed that females spent more time sniffing and in proximity to scents from the most distantly related males, but female response was not influenced by male urine testosterone concentration, female parity, age, or estrous cycling. Further research will be necessary to determine whether a female's interest in male urine translates to mate preference and acceptance before this technique can be applied to zoo breeding programs.

Variable duration of reproductive suppression in male coyotes (Canis latrans) treated with a high dose of the gonadotrophin-releasing hormone agonist deslorelin.

Effective and humane management strategies for coyotes (Canis latrans) remain elusive. We hypothesised that exposure to a high dose of a gonadotrophin-releasing hormone (GnRH) agonist would cause prolonged suppression of the reproductive axis. Two groups of male coyotes were administered 47mg deslorelin in the form of either five 9.4-mg controlled-release Suprelorin (Peptech Animal Health, Macquarie Park NSW, Australia) implants (n=3) or 10 4.7-mg implants (n=5). In the first group, deslorelin suppressed plasma LH, testosterone and testes volume in two of three coyotes for three breeding seasons. In the second group, two of five deslorelin-treated coyotes had no sperm production after 1 year and plasma LH, FSH, testosterone and testes volume were suppressed. Although plasma gonadotropins and testosterone were suppressed in three treated coyotes in group two, testes volume and sperm production were evident. Because the duration of suppression differed among individual coyotes, we further hypothesised that a variation in deslorelin release underlay the variability. To test this, we analysed in vivo plasma profiles of deslorelin concentrations. These profiles suggested that deslorelin concentrations >100 pg mL-1 are required to maintain suppression in male coyotes. For field implementation, the development of an implant capable of releasing deslorelin for the life of the coyote is necessary.

Parent-offspring behavior of Jambu fruit doves (Ptilinopus jambu).

Fruit doves (Ptilinopus) constitute a genus of small to medium-sized, brightly colored arboreal birds, whose diets consist entirely of fruit. Little is known about the behavior of fruit doves because most species inhabit dense forests and are difficult to observe in the wild. This study describes the parental behavior of Jambu fruit dove pairs (Ptilinopus jambu) in a captive breeding program at the Saint Louis Zoo. Continuous video recordings were made of three pairs which raised a total of eight squabs over 2 years; daily rates of parental and squab behaviors were quantified. Overall, females were present at the nest, brooded their squabs, pecked, and attempted feedings more often than males. Parents also cared for their squabs at different times throughout the day. Males fed and brooded squabs during the middle of the day, while females fed throughout the day and brooded in the morning, evening, and overnight. Feeding rates were lower than those described for seed-eating doves (Columbinae), with hours between consecutive feedings, and squabs rarely begged before feeding events. Most squab behaviors involved initiating or terminating brooding and self-preening. These behaviors increased as squabs approached fledging, and coincided with a shift from full to partial brooding, and a decrease in parental allopreening. Older squabs also initiated feeding less frequently. Together, these data provide the first description of parental behavior in a Ptilinopus fruit dove. The results of this study may help improve captive breeding efforts, which are likely to become increasingly important for future conservation and reintroduction programs.

A survey of bonobo (Pan paniscus) oral contraceptive pill use in North American zoos.

Contraception is an essential tool in reproductive management of captive species. The Association of Zoos and Aquariums (AZA) Reproductive Management Center (RMC) gathers data on contraception use and provides recommendations. Although apes have been given oral contraceptive pills (OCPs) for at least 30 years, there have been no published reports with basic information on why the pill is administered, formulations and brands used, and effects on physiology and behavior. Here, we report survey results characterizing OCP use in bonobos (Pan paniscus) housed in North American zoos, as well as information accumulated in the RMC's Contraception Database. Of 26 females treated, there have been no failures and nine reversals. The most commonly administered OCP formulation in bonobos contained ethinyl estradiol (EE) 35 µg/norethindrone 1 mg. Few females on combined oral contraceptives (COCs) were given a continuous active pill regimen; a hormone-free interval of at least 5 days was allowed in most. Crushing the pill and mixing with juice or food was common. Females on COCs seldom experienced breakthrough estrus or bleeding, while these conditions were sometimes observed for females on continuous COCs. All females on COCs exhibited some degree of perineal swelling, with a mean score of 3 or 3+ most commonly reported. Behavioral changes included less sexual behavior, dominant females becoming subordinate, and a negative effect on mood. No appreciable change in weight was noted. Taken together, these results indicate that OCPs are an effective and reversible contraceptive option for bonobos that can be used by zoos and sanctuaries to limit reproduction. Zoo Biol. 35:444-453, 2016. © Wiley Periodicals, Inc.

Reproductive behavior of the great hornbill (Buceros bicornis).

Great hornbills (Buceros bicornis) are a long-lived, monogamous species that forms strong pair-bonds, and mate compatibility is thought to be important for successful reproduction. Within AZA, great hornbills are listed as a red SSP. The population consists of a limited number of individuals that do not breed reliably, and improving reproduction is a top priority for the Coraciiformes TAG. To better understand mating behavior and evaluate mate compatibility, this study documented the behavior of pairs of great hornbills during and immediately after courtship. Using live observations, the study followed one female, an experienced and successful breeder, as she was paired with four successive males over 11 breeding seasons. Initially, males frequently vocalized, investigated the nest, and approached the female. As the female spent more time in the nest, these behaviors were replaced by regurgitation and food offering. The female was most often observed plastering and vocalizing. Behavioral differences between successful and unsuccessful pairs, possibly indicative of pair compatibility, included rates of approaching, billing, and biting. Numerous behaviors occurred more frequently during years that a chick hatched, including pseudoregurgitation, regurgitation, offering food items, and nest investigation. Males also spent more time in proximity to both the female and the nest during years that a chick hatched. Together, these results suggest that the amount of time pairs spend in proximity, the amount of time a male spends near the nest, and the frequency of certain behaviors may help evaluate compatibility and the likelihood of successful reproduction for pairs of great hornbills.

"Use it or lose it": characterization, implications, and mitigation of female infertility in captive wildlife.

Zoos and other ex situ wildlife institutions can play an important role in species conservation by maintaining populations for education and research, as sources for potential re-introduction or reinforcement, and as ambassadors for financial support of in situ conservation. However, many regional zoo associations are realizing that current captive populations are unsustainable, with many programs failing to meet demographic and genetic goals to ensure long-term viability. Constraints on population size due to limited space often mandate delayed and/or less frequent breeding, but for females of many species this can have profound effects on fertility. A retrospective analysis combined with published literature and reliable anecdotal reports reveals that, when females are housed in a non-breeding situation for extended periods of time, reproductive changes that negatively impact fertility have occurred in multiple species, including canids, elephants, white rhinoceros, Seba's bats, wildebeest, stingrays, and some felid species. Competing space needs and changing interest in taxa for exhibits over time compound the problem. Counter strategies to breed early and often have their own demographic and genetic consequences as well as logistical and political implications. Strategies to mitigate the sustainability crisis in these taxa might include a mixed strategy in which young, genetically valuable females are bred earlier and at more regular intervals to ensure reproductive success, in combination with the judicious use of available tools to manage the number of offspring produced, including contraception and culling. An understanding of the issues at stake is the first step towards developing management strategies for sustainable populations.

Factors associated with uterine endometrial hyperplasia and pyometra in wild canids: implications for fertility.

The ability to safely and effectively manage reproduction is central to the success of AZA captive-breeding programs. Although the AZA Wildlife Contraception Center routinely monitors contraceptive safety, there have been no studies that compare the effects of contraceptive use to separation of males from females, the other option for preventing reproduction. We used retrospective medical records and pathology reports submitted by AZA and related facilities for the seven AZA-managed canid species to assess rates of uterine pathology relative to female reproductive life histories. Our results showed that the prevalence of both pyometra and endometrial hyperplasia (EH) was associated not only with treatment with the two most common contraceptives (Suprelorin® and MGA implants) but also with the number of years barren (i.e., not producing a litter and not contracepted). Rates of pyometra and EH were especially high in African painted dogs and red wolves, but lowest in swift and fennec foxes. The number of years producing a litter had a low association, suggesting it could be protective against uterine pathology. A more recently developed Suprelorin® protocol using Ovaban® to prevent the initial stimulation phase, followed by implant removal when reversal is desired, may be a safer contraceptive option. These results concerning the relationship between reproductive management and uterine health have important implications for AZA-managed programs, since the unsustainability of many captive populations may be due at least in part to infertility. Managing a female's reproductive lifespan to optimize or maintain fertility will require a reconsideration of how breeding recommendations are formulated.

Affiliative and aggressive behavior in a group of female Somali wild ass (Equus africanus somalicus).

We observed a group of three young female Somali wild asses to develop an ethogram of social behavior in the first phase of a longer term study of social, sexual, and maternal/infant behavior. The most unexpected finding was the frequency and extent of aggressive interactions, which included Charge, Drive, Neck Wrestle, Head Butt, and Body Slam, behaviors previously reported only for males of other equid species. The overall frequency of aggressive behavior was higher than that of affiliative behavior (84±16.5 vs. 32±5.5, P=0.03), yet no injuries occurred. The dyadic directionality of aggressive behavior suggested a dominance hierarchy, a feature not previously reported for either wild ass or domestic donkeys. The aggression observed may be an accurate representation of the behavior of this species, or their relatively young ages, or their recent transfer from their natal group through quarantine and into a new enclosure may have heightened agonistic tendencies. Further studies will determine whether with time their aggressive behavior becomes more intense or dissipates with maturity.

Nutrition and behavior of fennec foxes (Vulpes zerda).

Fennec foxes make popular pets because of their small size, minimal odor, and highly social behaviors. They are kept in zoos for conservation and educational programs. The exotic animal practitioner is most likely to be presented with fennec foxes that are overweight because of inappropriate diets or excessive feeding. Clients attempting to hand-rear fennec foxes need advice about formula selection, amounts to feed, protocols for keeping pups warm, and weaning. This article provides information on social behavior, reproduction, and parental behavior, nutrition, and hand-rearing.

Scent and the single male: ring-tailed lemurs produce honest signals.

In mating systems that involve competing males and choosy females, males are expected to advertise their genetic quality to discriminating females. Most examples have focused on visual or acoustic signals, such as ornamentation or song; yet arguably, olfactory communication may be more important to the majority of vertebrates with the possible exception of birds. Fortunately, attention has begun to shift to the role of odours in mate choice, with most of that attention being directed at the major histocompatibility complex or more recently at the major urinary proteins. The study of male ring-tailed lemurs presented by Charpentier and colleagues in this issue adds a new dimension to investigations of the influence of genes on mate choice via odour production. By comparing genetic heterozygosity to the production of semiochemicals in the scrotal scent gland, they provide a link between genetic composition and scent-marking behaviour as a potential advertisement of male quality.

Pregnancy diagnosis in wild canids using a commercially available relaxin assay.

Accurate detection of pregnancy is a useful tool in zoo management and husbandry, conservation breeding programs and research settings. Our study evaluated the ability of a commercial relaxin hormone assay used in domestic dogs (ReproCHEK(TM)) to accurately detect pregnancy through plasma analysis in two wolf and two fox species. The relaxin assay detected all of the pregnancies greater than 25 days gestation for island foxes, fennec foxes, gray wolves, and Mexican gray wolves. For island foxes, three negative relaxin results were attributed to using the test earlier postconception than manufacturer recommendation (before day 20). Five other negative results were found for females estimated at 15-25 days gestation, spanning the early and intermediate period (21-30 days gestation) when relaxin may be detected but less reliably. There were no false-positive results in nonmated negative control animals. Relaxin assay results were highly correlated with ultrasound results and the intra-assay replicate agreement was 100%. Our results show that the ReproCHEK(TM) commercial relaxin assay is a minimally invasive and reliable method for pregnancy detection in these wild species when used after 25 days gestation and might be applied to other canids as well. Furthermore, this assay is easy to run and requires no specialized equipment, making it extremely useful for zoo and field research applications. Zoo Biol 27:406-413, 2008. (c) 2008 Wiley-Liss, Inc.

Contraception in Dogs and Cats.

Although surgical sterilization is the most common approach to reproductive management of dogs and cats in North America and many other places, there is growing interest in nonsurgical and reversible methods. This review summarizes the methods currently available for use as reversible contraception in dogs and cats, along with cautions about reported side effects. The products covered include synthetic steroid hormones, peptide hormone agonists, vaccines, and a barrier method.

Reproductive cycles and contraception of black lemurs (Eulemur macaco macaco) with depot medroxyprogesterone acetate during the breeding season.

Contraception is a critical component of population management for lemurs, but concerns about potential deleterious effects of continuous, long-term treatment with synthetic progestins such as the widely used melengestrol acetate implant led us to evaluate seasonal contraception with injections of depot medroxyprogesterone acetate (Depo-Provera, Upjohn Pharmacia, Kalamazoo, MI) to limit the duration of exposure. We compared two dosage regimens in female black lemurs using vaginal cytology as an indirect measure of ovarian suppression. Our results indicate that both 10 mg/kg body weight at 90-day intervals or 2.5 mg/kg at approximately 30-day intervals can be effective in most females, although one female on the 10-mg dose showed signs of estrus at 53 days. Darkening of pelage during treatment was the primary side effect noted. A more important observation was that contraception can extend the breeding season to as much as 9 months, considerably longer than reported previously, which necessitates extending the period of contraceptive treatment. Zoo Biol 26:289-298, 2007. (c) 2007 Wiley-Liss, Inc.

Restoration of reproductive potential after expiration or removal of melengestrol acetate contraceptive implants in Tigers (Panthera tigris).

The need for contraception in the successful management of captive wild animals is becoming increasingly apparent. Because concerns exist regarding the reversibility of the contraceptive implant melengestrol acetate (MGA), reproductive data for 94 female Amur (Panthera tigris altaica) and Sumatran tigers (Panthera tigris sumatrae) were analyzed using survival analyses to evaluate return to reproductive status after implant removal or assumed expiration. Females placed in potential breeding situations after MGA implants were surgically removed showed a 62% return to reproduction by 5.25 years, whereas females with implants that were assumed to have expired showed only a 30% return to reproduction by 6 years. Implanted females did not reproduce as successfully as non-implanted control females, which showed an 85% probability of reproducing after placement in a new breeding situation by 2.66 years. Parturition increased the probability of reproducing in non-implanted females, but not in implanted females. Litter size, stillbirths, and offspring survival were not significantly different between non-implanted, implant-removed and implant-expired female tigers. Ten female tigers reproduced both before and after implant placement, and the differences in litter size, stillbirths, and offspring survival were not significant, nor were they significantly different from non-implanted females. Prior parturition, age when implant was removed, and duration of implantation did not affect the probability of reproducing for females after implant removal. These results show substantial reversibility of MGA implants, leading to 62% successful reproduction after implant removal. The reasons for lower successful reproduction in animals previously treated with the contraceptive compared to non-implanted females are not known, but a greater delay in reversibility was seen when implants were left in place and only presumed expired. Zoo Biol 26:275-288, 2007. (c) 2007 Wiley-Liss, Inc.