Appeasing Pheromones for the Management of Stress and Aggression during Conservation of Wild Canids: Could the Solution Be Right under Our Nose?

Thirty-six species of canid exist globally, two are classified as critically endangered, three as endangered, and five as near threatened. Human expansion and the coinciding habitat fragmentation necessitate conservation interventions to mitigate concurrent population deterioration. The current conservation management of wild canids includes animal translocation and artificial pack formation. These actions often cause chronic stress, leading to increased aggression and the suppression of the immune and reproductive systems. Castration and pharmaceutical treatments are currently used to reduce stress and aggression in domestic and captive canids. The undesirable side effects make such treatments inadvisable during conservation management of wild canids. Pheromones are naturally occurring chemical messages that modulate behaviour between conspecifics; as such, they offer a natural alternative for behaviour modification. Animals are able to distinguish between pheromones of closely related species through small compositional differences but are more likely to have greater responses to pheromones from individuals of the same species. Appeasing pheromones have been found to reduce stress- and aggression-related behaviours in domestic species, including dogs. Preliminary evidence suggests that dog appeasing pheromones (DAP) may be effective in wild canids. However, the identification and testing of species-specific derivatives could produce more pronounced and beneficial behavioural and physiological changes in target species. In turn, this could provide a valuable tool to improve the conservation management of many endangered wild canids.

Dog appeasing pheromone prevents the androgen surge and may reduce contact dominance and active submission after stressful interventions in African wild dogs (Lycaon pictus).

The endangered African wild dog (AWD; Lycaon pictus) is a highly social canid living in packs with a separate male and female hierarchy. Immobilisation, handling and translocations are acute stressors for AWDs, however such interventions are often needed for species management. In addition, new pack formation or temporary pack separation can lead to an increase in intra-pack aggression. The goal of this double-blinded placebo-controlled study conducted in captive zoo populations was to evaluate whether dog appeasing pheromone (DAP) reduces behavioural stress and faecal glucocorticoid metabolite levels (fGCM) normally associated with pack separation, immobilisation and reintroduction (SIR), and to assess whether this reduces aggressive behaviours and faecal androgen metabolite levels (fAM). Four packs (n = 11 males) were treated with DAP and 4 packs (n = 12 males) were treated with a placebo solution, applied at the end of anaesthesia. Behavioural interactions as well as fGCM and fAM were determined from 3 days before until 4-6 days after SIR. No effect of DAP on fGCM was observed, however, fAM increased after SIR in placebo but not DAP treated animals. Moreover, on the day of reintroduction, DAP treated packs tended to have lower rates of contact-dominance and active-submission behaviour, but higher rates of non-contact dominance behaviour. As these effects could decrease the risk of agonistic interactions, DAP may be a useful tool to help manage new pack formations and temporary pack separation.

Social rank does not affect sperm quality in male African wild dogs (Lycaon pictus).

Sperm banking and AI could benefit endangered African wild dog conservation. However, it is unclear whether their dominance hierarchy causes a decrease in reproductive and sperm quality parameters in subordinate males that typically do not breed. In this study, we investigated the effect of social rank on male reproductive parameters, including faecal androgen and glucocorticoid metabolite concentrations, prostate and testes volume, preputial gland size, semen collection success and sperm quality. Samples were obtained from captive males (prebreeding season: n=12 from four packs; breeding season: n=24 from seven packs) that were classified as alpha (dominant), beta or gamma (subordinates) based on the frequency of dominant versus submissive behaviours. In the prebreeding season, semen was successfully collected from all alpha but only half the subordinate males, with urine contamination (associated with lower rank) significantly reducing total and progressive motility, sperm motility index, normal sperm morphology and acrosome integrity. The breeding season was associated with a significant increase in faecal androgens, prostate and testis volume, as well as progressive motility and the total number of spermatozoa ejaculated. However, with the exception of prostate volume (mean±s.e.m: 12.5±4.5, 7.1±1.0 and 7.3±1.0cm3 in alpha, beta and gamma males respectively; P=0.035), all other reproductive and sperm quality parameters did not differ between males of each social rank. In conclusion, reproductive suppression of subordinate males appears to be behaviourally mediated, because males of all social ranks produce semen of similar quality, making them suitable candidates for sperm banking, particularly during the breeding season when sperm quality improves.

Characterizing the reproductive biology of the female pygmy hippopotamus (Choeropsis liberiensis) through non-invasive endocrine monitoring.

The pygmy hippopotamus (Choeropsis liberiensis) is endangered in the wild and very little is known about its reproductive biology. In zoological facilities, this species experiences a number of reproductive issues that complicate breeding management, including a high rate of stillbirths and failure of many pairs to reproduce. We conducted a comprehensive study to evaluate reproductive cycles and pregnancy in this species using enzyme immunoassays (EIAs) for fecal hormone metabolite analysis. Fresh fecal samples were collected twice weekly for a one to three year period from 36 female pygmy hippos housed at 24 zoological institutions. Samples were analyzed in three separate laboratories. Three progestogen metabolite EIAs (Pg-diol: 5ß-pregnane-3α,20α-diol 3HS:BSA; PdG: pregnanediol-3-glucuronide R13904; mono-P4: Quidel clone 425) and three estrogen metabolite EIAs (E2a: estradiol-17ß-OH 17-HS:BSA; E2b: estradiol 17ß R0008; E2c: estradiol 17ß R4972) accurately reflected reproductive events. Average estrous cycle length was 31.8 ± 7.4 days based on estrogen metabolite peaks and 30.9 ± 7.3 days based on nadir to nadir progestogen metabolite concentrations. Cyclical patterns in both estrogen and progestogen metabolites were detected throughout the year, indicating a lack of seasonality. Estrogen metabolite peaks were also observed during pregnancy and lactation, suggesting that follicular development occurs during both reproductive states. Pregnancy was most reliably demonstrated by elevation in progestogen metabolites (Pg-diol or PdG) in the second half of gestation. Average gestation length based on breeding to calving date was 203 ± 4 days for 15 pregnancies. This comprehensive overview of the reproductive biology of the female pygmy hippo provides valuable data for guiding long-term breeding management for this endangered species and serves as a baseline for future studies addressing the potential influence of social structure, diet, body condition, and other husbandry factors on estrous cycling and reproduction.

GLP-2 administration results in increased proliferation but paradoxically an adverse outcome in a juvenile piglet model of short bowel syndrome.

OBJECTIVE: The objective of the present study was to examine the effect of glucagon-like peptide-2 (GLP-2) administration in a piglet, juvenile model of short bowel syndrome. MATERIALS AND METHODS: Four-week-old piglets underwent either a sham operation or 75% small bowel resection. Postoperatively, piglets received either polymeric infant formula diet or the diet and subcutaneous human recombinant GLP-2 (1600 microg/day for 7 days, 800 microg/day thereafter). Food intake was monitored throughout the experiment, and stool and serum samples obtained fortnightly. After the piglets were killed, tissues were obtained from the duodenum, jejunum, ileum, and terminal ileum, and used for morphological and functional analysis. RESULTS: Treatment with GLP-2 resulted in significantly increased numbers of proliferating and apoptotic cells in the ileum of sham and small bowel resection piglets (P < 0.05). GLP-2 administration resulted in decreased weight gain, serum albumin, and disaccharidases in both sham and small bowel resection piglets (P < 0.001 compared with polymeric infant formula diet alone). CONCLUSIONS: This is the first study to our knowledge to examine the effect of GLP-2 administration in a juvenile short bowel syndrome model. Contrary to adult rodent studies, administration of GLP-2 resulted in adverse outcomes including reduced ability to gain weight; decreased serum albumin, tissue maltase, and sucrase; and villous atrophy. We anticipate this information will have important implications for future paediatric clinical trials.

A perspective on the role of emerging technologies for the propagation of companion animals, non-domestic and endangered species.

Assisted reproductive technologies (ART) have been used successfully in humans, domestic and laboratory species for many years. In contrast, our limited knowledge of basic reproductive physiology has restricted the application of ART in companion animal, non-domestic and endangered species (CANDES). Although there are numerous benefits, and in some cases a necessity, for applying ART for the reproductive and genetic management of CANDES, the challenges encountered with even the most basic procedures have limited the rate of progress. In this foreword we discuss the status of conventional ART, such as artificial insemination and in vitro fertilisation, as well as their benefits and inherent difficulties when applied to CANDES. It is upon these techniques, and ultimately our knowledge of basic reproductive physiology, that the success of emerging technologies, such as those described in this special issue, are dependent for success.

Ovarian and testicular tissue xenografting: its potential for germline preservation of companion animals, non-domestic and endangered species.

In the present paper we aim to review the development of both ovarian and testicular xenografting with specific emphasis on its usage for companion animals, non-domestic and endangered species. Ovarian and testicular tissue xenografting has been used successfully across a variety of species for the harvesting of mature gametes and subsequent fertilisation. It has become a novel and promising tool to explore various aspects of testicular development and function and was useful for determination of gonadotoxic treatments on xenografted gonads. In rodent animal models live births have been reported using gametes from xenografted gonadal tissue. Live births were also reported after grafting of human ovarian tissue. We envisage that it will not be long before the first live births across other species, including companion animals, non-domestic and endangered species, will be achieved.

Preservation of female germ cells from ovaries of cat species.

This review provides an overview on recent knowledge on female germ cell population within cat ovaries; on isolation, culture and cryopreservation of feline preantral follicles and on ovarian tissue preservation.

Colostrum protein concentrate enhances intestinal adaptation after massive small bowel resection in juvenile pigs.

OBJECTIVES: Short bowel syndrome (SBS) usually results from the surgical removal of a large segment of small intestine. Patient outcome depends on the extent of intestinal resection and adaptation of the remaining intestine. We evaluated the impact of colostrum protein concentrate (CPC) on intestinal adaptation after massive small bowel resection in a porcine model of infant SBS. METHODS: Four-week-old piglets underwent an approximate 75% small bowel resection (R, n = 23) or a control transection operation (C, n = 14). Postoperatively, animals from both groups received either pig chow (R = 6, C = 5), polymeric infant formula (R = 6, C = 3) or polymeric infant formula supplemented with CPC (R = 11, C = 6) for 8 weeks until sacrifice. Clinical outcome measures included weight gain and stool consistency. Morphologic measures were intestinal villus height and crypt depth. Functional outcome measure was mucosal disaccharidase activity. RESULTS: Resected animals fed polymeric infant formula alone had reduced weight gain compared with controls fed the same diet (P < 0.005). Despite massive small bowel resection, animals fed pig chow or polymeric infant formula supplemented with CPC grew at an equivalent rate to controls fed polymeric infant formula alone. Resected animals supplemented with CPC had increased villus length and crypt depth in the jejunum (P < 0.001) and ileum (P < 0.001) compared with resected animals fed either pig chow or polymeric infant formula alone. CONCLUSION: In an animal model of SBS, CPC supplementation of polymeric infant formula resulted in normal weight gain and features of enhanced morphologic adaptation.

Xenotransplantation: a tool for reproductive biology and animal conservation?

The transplantation of reproductive organs, including ovaries and ovarian tissue, was pioneered over 100 years ago. In the 1960s, ovarian grafting was used as a tool to investigate ovarian function, but with the recent development of more effective cryopreservation protocols for ovarian tissue, germline preservation and propagation have now become realistic goals. This review describes progress in ovarian banking and ovarian tissue transplantation, with emphasis on how fresh and frozen ovarian tissue can be used in assisted reproduction for both humans and animals. This paper focuses most closely on the potential value of xenotransplantation, the transplantation of gonads from one species to another, to conserve rare and endangered species. Specific attention is drawn to the use of xenotransplantation as a strategy for generating viable gametes that can be used to produce live fertile offspring. Other upcoming xenogeneic technologies that may be of potential significance in animal conservation, such as transplantation of whole ovaries or isolated growing follicles, and even male germ cells, are discussed.

Influence of diet complexity on intestinal adaptation following massive small bowel resection in a preclinical model.

AIMS: To investigate the effect of dietary complexity on intestinal adaptation using a preclinical model. METHODS: Four-week-old piglets underwent a 75% proximal small bowel resection or transection operation (control). Post-operatively, animals received either pig chow (n = 15), polymeric formula (n = 9), polymeric formula plus fiber (n = 6), or elemental formula (n = 7). RESULTS: The weight gain of all groups was reduced compared with controls that were fed the same diet. Animals that had a resection, which were fed elemental formula, had significantly reduced weight gain compared with the other groups (4.7 4.2 vs 30.7 7.1 kg chow and 11.5 1.3 kg polymeric formula). Villus height was increased in the jejunum, ileum and terminal ileum of resected animals compared with controls in animals fed with pig chow, polymeric formula and elemental formula. The animals that had a resection had a significant reduction in the transepithelial conductance (10.4 5.5 vs 25.4 6.5 mS/cm2) and 51Chromium-EDTA flux (2.8 1.9 vs 4.8 4.9 microL/h per cm2) compared with the controls. CONCLUSIONS: A complex diet was found to be superior to an elemental diet in terms of the morphological and functional features of adaptation following massive small bowel resection.