Linking social environment and stress physiology in feral mares (Equus caballus): group transfers elevate fecal cortisol levels.

Feral horses (Equus caballus) have a complex social structure, the stability of which is important to their overall health. Behavioral and demographic research has shown that decreases in group (or band) stability reduce female fitness, but the potential effects on the physiological stress response have not been demonstrated. To fully understand how band stability affects group-member fitness, we need to understand not only behavioral and demographic, but also physiological consequences of decreases to that stability. We studied group changes in feral mares (an activity that induces instability, including both male and female aggression) on Shackleford Banks, NC. We found that mares in the midst of changing groups exhibit increased fecal cortisol levels. In addition, mares making more group transfers show higher levels of cortisol two weeks post-behavior. These results offer insights into how social instability is integrated into an animal's physiological phenotype. In addition, our results have important implications for feral horse management. On Shackleford Banks, mares contracepted with porcine zona pellucida (PZP) make approximately 10 times as many group changes as do untreated mares. Such animals may therefore be at higher risk of chronic stress. These results support the growing consensus that links between behavior and physiological stress must be taken into account when managing for healthy, functional populations.

A free-ranging, feral mare equus caballus affords similar maternal care to her genetic and adopted offspring.

Adoption of nongenetic offspring occurs in a variety of species but is rare in equids. We report a case of adoption by a free-ranging, feral mare Equus caballus and compare the maternal care received by her genetic offspring (born 1995) to that of her adopted offspring (born 1996) for the first 30 weeks of development. We compare five measures of care: (1) total time spent suckling, (2) mare aggression during suckling, (3) number of mare-terminated suckling bouts, (4) contact maintenance, and (5) mare-foal distance. For most behaviors, we detected no difference in the mare's treatment of the two foals; however, mare-foal distance was greater for the genetic offspring. We compare hypotheses regarding the reasons for adoption, offering postpartum physiological state as a potential driver.

Laissez-Faire Stallions? Males' Fecal Cortisol Metabolite Concentrations Do Not Vary with Increased Female Turnover in Feral Horses (Equus caballus).

Stress responses can be triggered by several physical and social factors, prompting physiological reactions including increases in glucocorticoid concentrations. In a population of feral horses (Equus caballus) on Shackleford Banks, North Carolina, females previously immunized with the immunocontraceptive agent porcine zona pellucida (PZP) change social groups (bands) more often than unimmunized females, disrupting the social stability within the population. We assessed the effects of increased female group changing behavior (or female turnover) on individual male stress by comparing fecal cortisol metabolite (FCM) concentrations among stallions experiencing varying amounts of female group changing behavior. FCM concentrations did not significantly correlate with female turnover. Similarly, FCM concentrations were not dependent upon the timing of female group changing behavior. These findings suggest that female turnover rate has little influence on physiological measures of stress in associated stallions. That said, Shackleford stallions experiencing increased female turnover do engage in behaviors typically associated with stress (increased vigilance, highly escalated male-male conflicts). Future work should compare FCM concentrations across time within populations and among populations managed under different strategies to better isolate factors influencing stallion stress physiology. Such studies are especially important if we are to determine how changes in female behavior related to immunocontraception impact physiological and behavioral indicators of stress for non-target animals. Finally, our study highlights the importance of considering both physiological and behavioral measures when investigating animal responses to potentially challenging situations.

Out of the stable: Social disruption and concurrent shifts in the feral mare (Equus caballus) fecal microbiota.

The disruption of animals' symbiotic bacterial communities (their microbiota) has been associated with myriad factors including changes to the diet, hormone levels, and various stressors. The maintenance of healthy bacterial communities may be especially challenging for social species as their microbiotas are also affected by group membership, social relationships, microbial transfer between individuals, and social stressors such as increased competition and rank maintenance. We investigated the effects of increased social instability, as determined by the number of group changes made by females, on the microbiota in free-living, feral horses (Equus caballus) on Shackleford Banks, a barrier island off the North Carolina coast. Females leaving their groups to join new ones had fecal microbial communities that were similarly diverse but compositionally different than those of females that did not change groups. Changing groups was also associated with the increased abundance of a several bacterial genera and families. These changes may be significant as horses are heavily dependent upon their microbial communities for nutrient absorption. Though we cannot identify the particular mechanism(s) driving these changes, to the best of our knowledge, ours is the first study to demonstrate an association between acute social perturbations and the microbiota in a free-ranging mammal.

Communication is key: Mother-offspring signaling can affect behavioral responses and offspring survival in feral horses (Equus caballus).

Acoustic signaling plays an important role in mother-offspring recognition and subsequent bond-formation. It remains unclear, however, if mothers and offspring use acoustic signaling in the same ways and for the same reasons throughout the juvenile stage, particularly after mutual recognition has been adequately established. Moreover, despite its critical role in mother-offspring bond formation, research explicitly linking mother-infant communication strategies to offspring survival are lacking. We examined the communicative patterns of mothers and offspring in the feral horse (Equus caballus) to better understand 1) the nature of mother-offspring communication throughout the first year of development; 2) the function(s) of mother- vs. offspring-initiated communication and; 3) the importance of mare and foal communication to offspring survival. We found that 1) mares and foals differ in when and how they initiate communication; 2) the outcomes of mare- vs. foal-initiated communication events consistently differ; and 3) the communicative patterns between mares and their foals can be important for offspring survival to one year of age. Moreover, given the importance of maternal activity to offspring behavior and subsequent survival, we submit that our data are uniquely positioned to address the long-debated question: do the behaviors exhibited during the juvenile stage (by both mothers and their young) confer delayed or immediate benefits to offspring? In summary, we aimed to better understand 1) the dynamics of mother-offspring communication, 2) whether mother-offspring communicative patterns were important to offspring survival, and 3) the implications of our research regarding the function of the mammalian juvenile stage. Our results demonstrate that we have achieved those aims.

Lingering effects of contraception management on feral mare (Equus caballus) fertility and social behavior.

Due to the extirpation of their natural predators, feral horse populations have expanded across the United States, necessitating their management. Contraception of females (mares) with porcine zona pellucida (PZP) is a popular option; however, effects to physiology and behavior can be substantial. On Shackleford Banks, North Carolina, USA, treated mares have exhibited cycling during the non-breeding season and demonstrated decreased fidelity to the band stallion, but PZP's long-term effects on mare physiology and behavior remain largely unexplored. After the contraception program was suspended in this population, we examined how prior exposure to varying levels of PZP treatment impacted (1) foaling probability and foaling dates (a proxy for ovulatory cycling) from 2009 to 2014 and (2) mare fidelity to the band stallion and reproductive behavior during 2013 and 2015. Additionally, we evaluated the effects of time since the mares' last treatment on these factors. Mares receiving any level of prior PZP treatment were less likely to foal than were untreated mares. Among mares that received 1-3 PZP applications, foaling probability increased with time since last treatment before declining, at ~6 years post-treatment. Mares that received 4+ applications did not exhibit a significant increase in foaling probability with time since last treatment. Moreover, previously treated mares continued to conceive later than did untreated mares. Finally, mares previously receiving 4+ treatments changed groups more often than did untreated mares, though reproductive behavior did not differ with contraception history. Our results suggest that although PZP-induced subfertility and its associated behavioral effects can persist after the cessation of treatment, these effects can be ameliorated for some factors with less intense treatment. Careful consideration to the frequency of PZP treatment is important to maintaining more naturally functioning populations; the ability to manage populations adaptively may be compromised if females are kept subfertile for extended periods of time.

Immunocontraception in wild horses (Equus caballus) extends reproductive cycling beyond the normal breeding season.

BACKGROUND: Although the physiological effects of immunocontraceptive treatment with porcine zona pellucida (PZP) have been well studied, little is known about PZP's effects on the scheduling of reproductive cycling. Recent behavioral research has suggested that recipients of PZP extend the receptive breeding period into what is normally the non-breeding season. METHODOLOGY/PRINCIPAL FINDINGS: To determine if this is the case, we compiled foaling data from wild horses (Equus caballus) living on Shackleford Banks, North Carolina for 4 years pre- and 8 years post-contraception management with PZP (pre-contraception, n = 65 births from 45 mares; post-contraception, n = 97 births from 46 mares). Gestation lasts approximately 11-12 months in wild horses, placing conception at approximately 11.5 months prior to birth. Since the contraception program began in January 2000, foaling has occurred over a significantly broader range than it had before the contraception program. Foaling in PZP recipients (n = 45 births from 27 mares) has consistently occurred over a broader range than has foaling in non-recipients (n = 52 births from 19 mares). In addition, current recipients of PZP foaled later in the year than did prior recipient and non-recipient mares. Females receiving more consecutive PZP applications gave birth later in the season than did females receiving fewer applications. Finally, the efficacy of PZP declined with increasing consecutive applications before reaching 100% after five consecutive applications. CONCLUSIONS/SIGNIFICANCE: For a gregarious species such as the horse, the extension of reproductive cycling into the fall months has important social consequences, including decreased group stability and the extension of male reproductive behavior. In addition, reproductive cycling into the fall months could have long-term effects on foal survivorship. Managers should consider these factors before enacting immunocontraceptive programs in new populations. We suggest minor alterations to management strategies to help alleviate such unintended effects in new populations.