Ethological investigations on a herd of plains zebra in a safari park: Time-budgets,reproduction and food competition

The social organization of a herd of 25 plains zebras under safari park conditions and the time-budgets of 18 individuals from this herd were investigated. The zebras had organized themselves in harem groups. Stallions appeared to spend more time acting socially than did other sex-age classes and they spent less time feeding. Social behaviours were mainly directed against other stallions. Stallions were also more on the alert. The time-budget of safari park zebras largely agrees with that of their wild counterparts. From data on time budgets and from differences in the reproductive output of adults of different ranks it is concluded that differential access to food plays an important role in the safari park.     

Female–female competition or male mate choice? Patterns of courtship and breeding behavior among feral horses (Equus caballus) on Assateague Island

Previous research on the Assateague horses found that high-ranking females had more surviving offspring than low-ranking females. Variance in reproductive success may be the result of a variety of proximate processes that affect sexual behavior such as mate choice and mate competition. A study was done to determine whether patterns of courtship, social, and sexual behavior could be identified that would suggest mate choice and/or mate competition. Behavioral data were collected from approximately 40 sexually mature mares living in harem bands. Stallions showed more interest in the eliminations of dominant mares than subordinate mares. Males also engaged in significantly more high-intensity (e.g., mounts and copulations) sexual behavior with dominant mares than subordinate mares, and there was a trend for males to engage in more low-intensity (e.g., flehmen and ano-genital sniffing) sexual behavior with dominant mares than subordinate mares. There was no effect of mare rank on spatial relationships with the stallion; however, dominant mares did attempt to restrict reproductive access to the stallion by harassing and disrupting copulations. Higher foaling rates among dominant mares on Assateague Island could therefore be the result of rank-related mate choice by stallions and direct female competition for mating opportunities.     

Sexual behavior following introduction of a stallion into a group of mares

A rested stallion was introduced daily for 30 days into each of three herds of 20 mares. Observations of sexual and mating behavior were made for one hour. The stallion remained with a herd until another stallion was introduced the following day. Other mares were isolated from stallions and were bred by artificial insemination. The diameter or growth rate of the preovulatory follicle for the six days preceding ovulation and the length of the interovulatory interval for mares which did not become pregnant were not affected significantly by the presence of a stallion. The number of breedings per hour of observation (2.4 ±0.2) and the length of the interval from introduction of a stallion into the herd of mares to first breeding (12 ±1 min) were significantly different among stallions, but the length of the interval between breedings (17 ±2 min) was not. The mean number of breedings per stallion per hour was not affected significantly by the number of posturing (estrous) mares. The number of times that the stallion rebred the same mare when more than one mare postured during the observation hour (49%) was greater (P<0.01) than what would be expected to occur by chance (30%). The hypothesis that breeding occurs preferentially in those estrous mares that are closest to ovulation was not supported, except for significantly lower breeding activity in posturing mares on days 8 and 7 before ovulation and on days 0 (day of ovulation) and ?1 (26% bred) than on days 6 to 1 (52%).     

Sociosexual behavior and the ovulatory cycle of ponies (Equus caballus) observed in harem groups

Observations of sociosexual behavior of adult ponies, made on two harem groups (each comprised of one vasectomized male and three females), were correlated with follicular development and ovulation for a total of 15 cycles (minimum of 2 cycles per female). Mean cycle length (interovulatory interval) was found to be 19.7 days, with behavioral estrus lasting 7–8 days (5.5 days preovulatory; 2.3 days postovulatory). Estrous females typically showed increased frequencies of approaching and following the stallion, urinating, presenting, clitoral winking, and tail raising. Approaching and following the stallion appeared earlier and persisted longer than other estrous responses. Deviations from the modal estrous pattern included cycles with subestrus, continual estrus, behavioral estrus in the absence of ovulation, and displays of female mounting. Dominance tests revealed that a mare's status was unaffected by the phases of the estrous cycle. The presence of more than one estrous female affected the copulatory performance of both stallions, most notably in reduced latencies to first mount, intromission, and ejaculation, in spite of differences between the stallions in sexual vigor. Each stallion usually selected the dominant mare for copulation when there were multiple estrous females present, but mounts were not displayed exclusively to one female per test. The social testing situation made apparent the importance of use of space in sociosexual communication in this species, particularly in avoidance of the stallion by diestrous mares and standing alone or in proximity to him by estrous mares.     

Density gradient centrifugation of sperm from a subfertile stallion and effect of seminal plasma addition on fertility

Stallions are not selected for fertility but for other criteria (pedigree, conformation, performances, progeny), therefore valuable but subfertile stallions with poor semen quality are frequently used in commercial breeding programs. The object of this study was to evaluate whether sperm selection through a silane-coated silica colloid gradient centrifugation, with or without the addition of seminal plasma of a high fertile stallion, could improve the pregnancy rates of an oligospermic valuable stallion in a commercial breeding program. In 2008 breeding season (experiment 1, n=104 mares), simple centrifugation and density gradient centrifugation of the sperm were compared. In 2009 and 2010 breeding seasons (experiment 2, n=125 mares), the effect of the addition of 5% seminal plasma to the extender after sperm selection was evaluated. In all mares deep horn uterine insemination was performed with 1 ml containing 50×10(6) morphologically normal progressive motile spermatozoa, 24-30 h after induction of ovulation with hCG. Pregnancy diagnosis by ultrasonography was performed 14 days following ovulation. Results showed a higher per cycle pregnancy rate (P>0.05) when sperm selection through a density gradient was used (62% vs. 42.3%, exp 1), while the addition of 5% seminal plasma did not influence the outcome (45.9% vs. 47.6%, exp 2) (P>0.05). An age-related decrease in the fertility of the stallion was observed when comparing the results from the different breeding seasons (P<0.05). In conclusion, sperm selection through a discontinuous density gradient enabled a normal per cycle pregnancy rate to be achieved from an oligospermic-subfertile stallion in a commercial breeding program, and no differences were observed regarding the addition of seminal plasma.     

Tests for cooperative behaviour between stallions

Breeding groups with multiple stallions occur sympatrically with single-stallion breeding groups in feral horse, Equus caballus, populations. Mutualism and reciprocal altruism between stallions have been proposed to explain the origin and functioning of multistallion bands. However, empirical support for these hypotheses is contradictory and incomplete. Furthermore, there are no explicit tests of the predictions that each hypothesis makes about stallion behaviour and social structure. We compared nine multistallion and 18 single-stallion bands in the Kaimanawa Ranges, New Zealand. Compared with agonistic behaviours, affiliative behaviours were relatively unimportant in the relationships between stallions within bands. The number of stallions in the band did not have a positive influence on mare group size, stability, home range quality or reproductive success in bands. Furthermore, there was a positive relationship between aggression ('intolerance') by the dominant towards subordinate stallions and the subordinates' effort in mare group defence ('helping') but a negative relationship between helping effort by subordinates and their proximity to, and mating with, the bands' mares. Therefore, the predictions of the mutualism and reciprocal altruism hypotheses were not supported. Indeed, for some of the predictions we found the opposite outcomes to be true. Multistallion bands had significantly poorer reproductive success, and dominant stallions were less tolerant of subordinates that helped most and reduced their access to mares. Nevertheless, in all other respects Kaimanawa stallions in multistallion bands behaved like those described elsewhere. Thus, we reject cooperative hypotheses for multimale breeding groups in horses and discuss the mate parasitism and consort hypotheses as better alternatives. Copyright 2000 The Association for the Study of Animal Behaviour.     

Exposure to stallion accelerates the onset of mares' cyclicity

Horses (Equus caballus) belong to the group of seasonally polyestrous mammals. Estrous cycles typically start with increasing daylight length after winter, but mares can differ greatly in the timing of onset of regular estrus cycles. Here, we test whether spatial proximity to a stallion also plays a role. Twenty-two anestrous mares were either exposed to one of two stallions (without direct physical contact) or not exposed (controls) under experimental conditions during two consecutive springs (February to April). Ovarian activity was monitored via transrectal ultrasound and stallion's direct contact time with each mare was determined three times per week for one hour each. We found that mares exposed to a stallion ovulated earlier and more often during the observational period than mares that were not exposed to stallions. Neither stallion identity nor direct contact time, mare age, body condition, size of her largest follicle at the onset of the experiment, or parasite burden significantly affected the onset of cyclicity. In conclusion, the timing of estrous cycles and cycle frequency, i.e., crucial aspects of female reproductive strategy, strongly depend on how the mares perceive their social environment. Exposing mares to the proximity of a stallion can therefore be an alternative to, for example, light programs or elaborated hormonal therapies to start the breeding season earlier and increase the number of estrous cycles in horses.     

Sexual behavior of mares

The mare is seasonally polyestrus, having an anovulatory period during the short light days of late fall and early winter, and beginning to ovulate as the days become longer during the winter. The complete estrus cycle is typically about 3 weeks, with 5 to 7 days of estrus and approximately 2 weeks of diestrus. When a mare lives within the natural social structure of the horse, i.e. a family band with several adult mares and one or more stallions, estrus is characterized by repeatedly approaching the stallion, frequent urination, deviating the tail away from the perineum, and standing still with the hind limbs spread apart. Diestrus is characterized by avoidance of an approaching stallion, and aggression toward the stallion, such as squealing, striking, and kicking, if he persists in attempting to court the diestrus mare. However, mares and stallions with long-term social relationships will often rest together, graze together and groom each other, all without sexual interactions. Hormonally, estrous behavior in the mare is initiated by estradiol that is secreted by the follicle, while estrous behavior is suppressed by progesterone, secreted by the corpus luteum. Mares are unusual among the ungulates in that they periodically exhibit estrous behavior during the anovulatory period. This is probably due to the release of estrogenic steroids secreted by the adrenal cortex. The display of sexual behavior by the mare throughout the year is thought to facilitate maintenance of the horse's social structure, in which the male remains with a group of females year round, in contrast with most ungulates in which the females and males only come together during the mating season.     

Pregnancy rates and sexual behavior under pasture breeding conditions in mares

Pony mares (n=480) and 16 stallions were assigned to four herds of 60 mares and one stallion (large herds) and to 12 herds of 20 mares and one stallion (small herds). The stallions remained with the herds continuously for all of the large herds and seven of the small herds. In the five remaining small herds the stallion was put into a herd for three hours every two days for 12 observation periods. Pregnancy rates and day of ovulation were estimated by size of embryonal enlargements. Mean pregnancy rates of 51% and 54% were obtained in the small herds and 42% in the large herds during a 48-day period (equivalent to two estrous cycles). Pregnancy rates for herds with the stallion present continuously were higher (P<0.01) for the small herds than for the large herds for days 1-24 (42% versus 19%). There was no effect of herd size on number of mares becoming pregnant per herd on days 1-24, but more mares (P<0.01) became pregnant during days 25-48 in the large herds (13.2 mares per herd versus 1.8). In the herds in which the stallion was present intermittently, the number of times that the stallion rebred the same mare when more than one mare was in estrus was greater (P<0.01) than what would be expected to occur by chance (observed, 21%; expected, 11%). Repeated breeding of the same mare seemed related to the availability or activity of the mare, since such mares more frequently followed and positioned themselves in the vicinity of the stallion. Most of the interferences by a mare which involved keeping the stallion and another mare apart were directed at the mare, whereas most of the interferences during mounting were directed at the stallion (P<0.01). Mares were more likely (P<0.01) to interfere when in estrus than when in nonestrus. When interfering mares were in nonestrus, their hostility was usually directed at the stallion (92%), whereas when in estrus their interference was more frequently directed at a mare (73%, P<0.01).     

Effect of feeding a DHA-enriched nutriceutical on the quality of fresh, cooled and frozen stallion semen

Eight stallions were used in 2 x 2 crossover study to determine if feeding a nutriceutical rich in docosahexaenoic acid (DHA) would improve semen quality. Stallions were randomly assigned to one of two treatment groups (n = 4 per group). Stallions were fed their normal diet (control) or their normal diet top-dressed with 250 g of a DHA-enriched nutriceutical. Feeding trials lasted for 14 week, after which a 14-week washout period was allowed and the treatment groups were reversed for another 14 week feeding trial. Feeding the nutriceutical resulted in a three-fold increase in semen DHA levels and 50% increase in the ratio of DHA to DPA in semen. Sperm motion characteristics in fresh semen were unaffected by treatment. After 24 h of cooled semen storage in an Equitainer, total and progressive motility did not differ between treatment groups, but sperm from stallions fed the nutriceutical exhibited higher velocity and straighter projectory (P < 0.05). After 48 h of cooled storage, increases in the percentages of sperm exhibiting total motility (P = 0.07), progressive motility (P = 0.06) and rapid motility (P = 0.04), were observed when stallions were being fed the nutriceutical. For a subset of four stallions, whose progressive sperm motility was <40% after 24 h of cooled storage when fed the control diet, feeding the nutriceutical resulted in improvements in mean progressive motility of sperm after 24 h (P = 0.10) and 48 h (P = 0.03) of storage. Feeding the nutriceutical resulted in similar improvements in motion characteristics being observed in frozen-thawed semen. While it appears that feeding the nutriceutical may improve the motion characteristics of cool-stored stallion semen, it may be most beneficial for stallions of marginal fertility whose sperm do not tolerate the rigors of cooling and storage. The nutriceutical also appeared to improve the freezability of semen. More dramatic improvements in semen quality may be observed if modifications in the main fat content of the diet are incorporated with the DHA supplement.     

The epidemiology of contagious equine metritis (CEM) in England 1977--1978).

Following an outbreak of CEM in England during 1977 a Code of Practice was introduced to control the disease in 1978. The Code recommended a bacteriological screening programme for Thoroughbred mares and stallions and improved standards of hygiene on the stud farm. As a result of the implementation of the Code a number of asymptomatic carrier mares was detected. Stallions which had transmitted CEM in 2977 and were treated did not transmit the disease during 1978. Two small outbreaks of CEM were reported during the 1978 breeding season.     

Testis size and onset of spermatogenesis in Cape mountain zebras (Equus zebra zebra)

Testis mass of adult Cape mountain zebra stallions (mean 70.0 g) was appreciably less than that of other zebra species and domestic horses. The histological appearance of the testes of 11-, 24- and 29-month-old colts was typically prepubertal. Spermatogenic activity of a 4-year-old stallion obtained at the end of summer was at a very low level, while a 4.5-year-old stallion obtained 6 weeks after the winter solstice showed a marked increase in spermatogenesis compared with the 4-year-old. Stallions 6.5-19 years of age collected in different seasons all showed active spermatogenesis.     

SNARE proteins and caveolin-1 in stallion spermatozoa: possible implications for fertility

Proteins implicated in the "SNARE hypothesis" for membrane fusion have been characterized in the acrosome of several mammalian species, and a functional role for these proteins during the acrosome reaction has been proposed. We have investigated the presence of SNAREs in equine sperm, using semen samples obtained from stallions with varying fertility. Immunocytochemical analysis revealed that members of different SNARE families can be detected on the acrosome of equine sperm, notably in the acrosomal cap and equatorial segment. These proteins include the t-SNARE syntaxin, the v-SNARE synaptobrevin/VAMP, the calcium sensor synaptotagmin, and the ATPase NSF. Also present is caveolin-1, a component of lipid rafts. Stallions with fertility problems presented the worst quality of sperm and acrosomal membrane, and had less sperm cells stained positively for SNAREs and caveolin-1, than sperm from fertile donors (p < 0.001). Ubiquitin surface staining was also performed and it seemed to inversely correlate with stallion fertility, supporting data obtained with the negative staining technique. A male-related problem was confirmed when mares that had failed to impregnate with samples from an infertile stallion were successfully inseminated with sperm from a fertile donor. Furthermore NSF, synaptotagmin and caveolin-1 staining seemed to be useful in predicting stallion fertility, i.e. significantly more sperm cells stained positively for these proteins in samples from fertile males. Although these results need to be expanded on a larger scale, they suggest that acrosomal and surface staining of equine sperm with novel probes may constitute useful tools in predicting stallion fertility.     

Hysteroscopic or rectally guided, deep-uterine insemination of mares with spermatozoa stored 18 h at either 5 degrees C or 15 degrees C prior to flow-cytometric sorting

Practical application of sex-selected spermatozoa in the horse industry would be greatly improved by the ability to develop simplified methods for shipping, storing, and inseminating sex-selected spermatozoa. Acceptable pregnancy rates have been achieved using fresh sex-sorted stallion sperm, however many stallion owners are reluctant to send their stallions to the sorter location for collection during the breeding season. Furthermore, the technology would be more applicable if the hysteroscopic insemination technique was not necessary for adequate pregnancy rates. Hysteroscopic insemination requires expensive equipment and specially trained personnel. In the present study, stallion sperm were sex-sorted after being stored at either 5 degrees C or 15 degrees C for 18 h. Twenty million sex-sorted sperm were then inseminated using one of two insemination techniques: the hysteroscopic method or the rectally guided, deep-uterine technique. Results were determined based on 16-day pregnancy status. A first-cycle pregnancy rate of 72% (18/25) was achieved when sperm were shipped at 15 degrees C, sex-sorted, and then inseminated using the hysteroscopic method. With these results, it can be concluded that stallions are not necessary at the sorter location to achieve acceptable fertility with sex-sorted sperm. There was a tendency for more mares to become pregnant when sperm were shipped at 15 degrees C prior to sorting, when compared to shipment at 5 degrees C. Similarly, there was a tendency for more mares to become pregnant when hysteroscopic insemination was utilized, when compared to the rectally guided, deep-uterine technique. These trends suggest that if larger group numbers were available, significant differences between the treatments may be revealed.     

Social relationships in a herd of Sorraia horses Part II. Factors affecting affiliative relationships and sexual behaviours

The influence of age, dominance rank, kinship and aggressiveness over affiliative relationships and sexual behaviours were analysed in a herd of Sorraia horses, Equus caballus, kept under extensive management. Subjects were 10 adult mares 5-18 years old that had known each other since birth, and a stallion introduced into the group for breeding for the first time. Kinship coefficient and dominance rank were the most important factors affecting affiliative relationships. Bonds were reciprocal and stronger among mares with higher kinship. Mares spent more time in proximity to close-ranking and lower-ranking females. Mares with stronger affiliative relationships or higher relatedness were not less aggressive towards each other. Affiliative relationships between the stallion and the mares were not reciprocal: lower-ranking mares formed stronger bonds with the stallion but he preferred the less genetically related mares for proximity. However, the stallion was involved in sexual behaviours more frequently with the mares that were more genetically related to him. These results suggest that kinship beyond close relatives may affect affiliative relationships both among familiar and among unfamiliar horses. However, the influence of kinship does not imply that horses possess a kin recognition system and alternative explanations are discussed.     

Contagious equine metritis--outbreak of the disease in Kentucky and laboratory methods for diagnosing the disease.

Contagious Equine Metritis (CEM) was initially reported during the 1977 breeding season in England (Crowhurst, 1977) and Ireland (Timoney, Ward & Kelly, 1977. The disease has also been diagnosed in France and Australia (Huges, Bryden & MacDonald, 1978). The first occurrence of CEM in the United States followed the importation or 2 stallions from France late in 1977 which resulted in an outbreak early in the 1978 breeding season (Swerczek, 1978). Mares usually develop clinical signs of CEM 8--10 days after being covered by an infected stallion, when a copious, greyish discharge is seen. Other mares may not show any outward signs of disease, but may have a shortened dioestrous period. Many mares recover spontaneously from the disease, but a small proportion become carriers of the CEM organism. The stallion does not show any clinical signs of disease, but remains a carrier. In this paper we recommend various laboratory procedures for the diagnosis of CEM in mares and stallions.     

Diagnosis and treatment of four stallions, carriers of the contagious metritis organism--case report

Contagious Equine Metritis (CEM), a venereal disease of horses caused by the bacterium Taylorella equigenitalis, was first diagnosed in 1977 and subsequently spread to many nations [Proc 24th AM Assoc Equine Pract (1979) 287]. The disease was confirmed in the United States in 1978 [Proc Am Assoc Equine Pract (1983) 295]. Specific regulatory procedures for this disease have been established in the United States and 37 other countries. From 1999 through 2001, four of 120 imported European stallions tested positive for CEM at a quarantine facility in Darlington, MD, USA. Two stallions were identified by positive bacterial cultures for T. equigenitalis on arrival. The other two positive stallions were negative on initial bacterial cultures, but were identified as CEM carriers when test mares (that they had mated) were culture-positive for T. equigenitalis. Since T. equigenitalis, is a fastidious slow-growing coccobacillus, additional sets of samples taken over a interval might be required to ensure positive stallions are detected before mating test mares. Likewise, additional sets of samples taken over a long interval after treatment of a stallion for CEM might be required to ensure that positive stallions treated for CEM are detected before mating test mares. Aggressive systemic antibiotic therapy accompanied by routine topical therapy might be required to treat some CEM-positive stallions.     

An abnormal facial gesture in an estrous mare

A 24-year-old mare exhibited abnormal behavior only when she was in estrus. She was subordinate to other mares. During diestrus she aggressed against an approaching stallion as a normal non-receptive mare would, but during estrus she flexed her limbs, clamped her tail and opened and closed her mouth when a stallion approached. The facial expression appeared when she first noticed the stallion or heard his neighs. The limb flexing occurred only when a stallion was in close proximity. The facial expression closely resembled snapping (also known as champing or tooth-clapping) of immature equids and jawing of receptive zebra and donkey mares.It is hypothesized that the mare's expression, the snapping of foals and the facial expression of donkeys and zebras are the same expression — one that indicates that the animal is in an approach—avoidance situation. In this case, the mare may have been both fearful of, and attracted to, stallions.     

Administration of oxytocin immediately after insemination does not improve pregnancy rates in mares bred by fertile or subfertile stallions

It is probable that reduced pregnancy rates in mares bred to subfertile stallions is attributable, in part, to the reduced number of normal spermatozoa that colonize the oviduct. Administration of oxytocin stimulates both uterine and oviductal contractility. The hypothesis that oxytocin may enhance sperm transport to/into the oviducts, and thereby increase pregnancy rates, was tested in 2 trials. For both trials, fertile estrous mares with follicles > or = 35 mm in diameter were inseminated once at 24 h after administration of 1500 to 2000 U hCG. The inseminate dose was limited to 100 million spermatozoa in order to lower pregnancy rates and thus increase the chance of detecting a treatment effect. Pregnancy status was determined by transrectal ultrasound examination 14 to 16 d after insemination. In Trial 1, 49 mares were inseminated with 4 mL extended semen from 1 of 3 stallions (1 fertile and 2 subfertile males). Immediately after insemination, the mares were administered either 20 U oxytocin or 1 mL saline intravenously. In Trial 2, 51 mares were inseminated with 4 mL extended semen from 1 of 4 stallions (1 fertile and 1 subfertile male used in Trial 1, and 2 additional fertile males). Immediately after insemination, and again 30 min later, mares were administered either 5 U oxytocin or 0.25 mL saline intramuscularly. To test for effects of treatment with oxytocin and for the interaction between semen quality and treatment, a generalized linear mixed regression model was used that accounted for the split-plot design (treatment within stallions), the random effect of stallion, the fixed effect of semen quality, the binary outcome of a single breeding trial, and the varying number of trials per stallion/treatment groups. Three treatment protocols or regimens were used: placebo, 5 U oxytocin injected twice intramuscularly, and 20 units oxytocin injected twice intravenously. Semen was classified as high (fertile stallions) or low (subfertile stallions) quality. No interaction between semen quality and treatment was detected (P > 0.10). The pregnancy rate of mares treated with oxytocin immediately after insemination was 30% (15/50) compared with 50% (25/50) for mares treated with saline immediately after breeding. Administration of oxytocin did not affect pregnancy rates (P > 0.10).     

Novel observation of play behaviour between a harem holder and a bachelor group of Przewalski’s horses in the wild

acta ethologica - Przewalski’s horses live in stable nonterritorial families (harem) of one or more harem stallions, several mares, and their offspring. The harem stallion typically behaves...