Seasonal serum concentrations of melatonin in cycling and noncycling mares

To determine whether secretory patterns of melatonin change throughout the seasons in mares, blood samples were drawn byvenipuncture from nine mares at noon and midnight for five successive days at monthly intervals from August through July at the University of Missouri in Columbia, MO. In addition, during September, December, March, and June, blood samples were drawn from indwelling catheters at 2-h intervals for 48 or 72 h. Mares were predominantly Quarter Horses weighing approximately 450 kg and ranged from 3 to 12 yr of age. Mares were housed in outdoor paddocks with three-sided run-in sheds for shelter. During the noon and midnight bleeding period, mares were placed in a larger open-sided barn with outside runs. Mares remained outdoors with the barn being used as a shelter in the event of inclement weather. All lights in the shed were converted to red light. Often, moonlight provided enough illumination to collect blood samples. Mares were returned to their normal paddock after each sampling period. For analysis of data, a mare was considered to be cycling if serum concentrations of progesterone were greater than 1 ng/ mL. For a mare to be classified as exhibiting a nocturnal rise of melatonin, serum concentrations of melatonin had to be at least two times greater at midnight than at noon. By month, a relationship did not exist (chi2; P > 0.05) among mares that were exhibiting estrous cycles and exhibiting nocturnal rises of melatonin. Likewise, examination of serum profiles of melatonin taken at 2-h intervals for 48 h revealed considerable variation among mares throughout the seasons. A nocturnal rise in serum melatonin was observed only in June (P < 0.02). In March and December, serum melatonin was greater in cycling mares than noncycling mares, but the elevation was not associated with light-dark periods (P < 0.01). Two of the mares exhibited estrous cycles throughout the seasons but melatonin secretion in these two mares were similar to that observed in the seven mares that demonstrated seasonal anestrous. From these results, it does not appear that changes in serum concentrations of melatonin are used as a cue to regulate cyclic activity in the mare throughout the seasons.     

The robustness of faecal steroid determination for pregnancy testing Kaimanawa feral mares under field conditions

AIMS: To investigate the utility of faecal oestrone sulphate (OS) concentrations for detecting pregnancy in mares during behavioural studies of feral horses, in which the collection and preservation of samples is not immediate. METHODS: Oestrone sulphate concentrations were measured in fresh dung samples collected from 153 free-roaming Kaimanawa mares throughout the year. In addition, multiple samples were taken from the same pile to investigate the reliability of diagnosis from a single sample, as well as the influence of time until preservation on OS concentrations. Samples were also taken before and after a 10mm simulated rainfall event to test for dilution of OS concentrations by rain. Oestrone sulphate concentrations in all samples were measured using an enzyme immunoassay. RESULTS: From approximately 150 to 250 days of gestation, OS concentrations were consistently >80 ng/g in mares which subsequently foaled. Mares which did not foal and had low faecal OS concentrations in multiple samples throughout the year had faecal OS concentrations of 31+/-13 ng/g (mean+/-s.d.) with an upper 95% confidence limit of 57 ng/g. Mares sampled from 1 week before to 1 month after behavioural oestrus, and that did not foal in the previous and subsequent seasons, had OS concentrations of 37+/-32 ng/g (mean+/-s.d.) with an upper 95% confidence limit of 100 ng/g. The standard error of oestrone sulphate concentrations in multiple samples from the same dung pile ranged from 1 to 37% of the mean. This large within-pile variation, however, did not result in incorrect diagnoses from single samples unless mares were within 18 days of parturition. Keeping samples at ambient temperatures for up to 16 hours did not affect OS concentrations. Simulated rainfall caused a 17% mean reduction in OS concentrations, but did not change pregnancy diagnoses. CONCLUSIONS: Faecal OS concentrations >100 ng/g were indicative of pregnancy in Kaimanawa mares. For mares more than 150 days post-mating, OS concentrations <57 ng/g were indicative of non-pregnancy, while concentrations between 57 and 100 ng/g provided an inconclusive diagnosis. A single sample from each dung pile collected within 16 hours of defecation was sufficient to accurately diagnose pregnancy in mares 150-250 days post conception. CLINICAL RELEVANCE: Measurement of OS concentrations in dung samples was a reliable and robust indicator of pregnancy status in feral mares 150-250 days post mating. This corresponds approximately to the period from May to August, given the seasonal breeding pattern in this population. This method of determining pregnancy status is suitable for field use in behavioural and demographic studies of wild horse populations.     

The robustness of faecal steroid determination for pregnancy testing Kaimanawa feral mares under field conditions

Aims: To investigate the utility of faecal oestrone sulphate (OS) concentrations for detecting pregnancy in mares during behavioural studies of feral horses, in which the collection and preservation of samples is not immediate.

Methods: Oestrone sulphate concentrations were measured in fresh dung samples collected from 153 free-roaming Kaimanawa mares throughout the year. In addition, multiple samples were taken from the same pile to investigate the reliability of diagnosis from a single sample, as well as the influence of time until preservation on OS concentrations. Samples were also taken before and after a 10mm simulated rainfall event to test for dilution of OS concentrations by rain. Oestrone sulphate concentrations in all samples were measured using an enzymeimmunoassay.

Results: From approximately 150 to 250 days of gestation, OS concentrations were consistently >80 ng/g in mares which subsequently foaled. Mares which did not foal and had low faecal OS concentrations in multiple samples throughout the year had faecal OS concentrations of 31 ±13 ng/g (mean ± s.d.) with an upper 95% confidence limit of 57 ng/g. Mares sampled from 1 week before to 1 month after behavioural oestrus, and that did not foal in the previous and subsequent seasons, had OS concentrations of 37 ± 32 ng/g (mean ± s.d.) with an upper 95% confidence limit of 100 ng/g.The standard error of oestrone sulphate concentrations in multiple samples from the same dung pile ranged from 1 to 37% of the mean. This large within-pile variation, however, did not result in incorrect diagnoses from single samples unless mares were within 18 days of parturition. Keeping samples at ambient temperatures for up to 16 hours did not affect OS concentrations. Simulated rainfall caused a 17% mean reduction in OS concentrations, but did not change pregnancy diagnoses.

Conclusions: Faecal OS concentrations > 100 ng/g were indicative of pregnancy in Kaimanawa mares. For mares more than 150 days post-mating, OS concentrations <57 ng/g were indicative of non-pregnancy, while concentrations between 57 and 100 ng/g provided an inconclusive diagnosis. A single sample from each dung pile collected within 16 hours of defecation was sufficient to accurately diagnose pregnancy in mares 150-250 days post conception.

Clinical Relevance: Measurement of OS concentrations in dung samples was a reliable and robust indicator of pregnancy status in feral mares 150-250 days post mating. This corresponds approximately to the period from May to August, given the seasonal breeding pattern in this population. This method of determining pregnancy status is suitable for field use in behavioural and demographic studies of wild horse populations.     

Seasonal and pulsatile dynamics of thyrotropin and leptin in mares maintained under a constant energy balance

The objective of this study was to determine if seasonal and/or pulsatile variations occur in plasma concentrations of thyrotropin (TSH) and leptin in mares while maintaining a constant energy balance. Blood samples were collected every 20 min during a 24 h period in winter and again in summer from six Quarter Horse type mares. Plasma concentrations of TSH, leptin, and T₄ were determined by radioimmunoassay. No differences were observed in body weight between winter (388.1 ± 12.5 kg) and summer (406.2 ± 12.5 kg; P = 0.11). Plasma concentrations of TSH were greater in the summer (2.80 ± 0.07 ng/ml) when compared to winter (0.97 ± 0.07 ng/ml; P < 0.001). Pulse frequency of TSH was not different between winter (6.17 ± 0.78 pulses/24 h) and summer (5.33 ± 0.78 pulses/24 h; P = 0.49). Mean TSH pulse amplitude, pulse area, and area under the curve were all greater in summer compared to winter (3.11 ± 0.10 ng/ml versus 1.20 ± 0.10 ng/ml, 24.86 ± 0.10 ng/ml min versus 13.46 ± 1.90 ng/ml min, 3936 ± 72.93 ng/ml versus 1284 ± 72.93 ng/ml, respectively; P < 0.01). Mean concentrations of leptin were greater in summer (2.48 ± 0.17 ng/ml) compared to winter (0.65 ± 0.17 ng/ml; P < 0.001). Pulsatile secretion patterns of leptin were not observed in any horses during experimentation. Mean concentrations of T₄ were greater in winter (20.3 ± 0.4 ng/ml) compared to summer (18.2 ± 0.4 ng/ml; P < 0.001). These seasonal differences between winter and summer provide evidence of possible seasonal regulation of TSH and leptin.     

Seasonal effects on the aerobic bacterial and fungal conjunctival flora of normal thoroughbred brood mares in Florida

Objective To evaluate seasonal effects on the presence or absence of fungal and aerobic bacterial flora of the conjunctival fornix of normal Florida Thoroughbred horses. Sample population Both eyes of 100 horses. Procedure Horses with normal anterior segment ophthalmic examinations from three farms in north central Florida were included. Each animal had the ventral conjunctival fornix of each eye swabbed with sterile culturettes. Samples were taken in October, January, April, and July (1999–2000). Aerobic and fungal cultures were plated. Bacterial cultures were reviewed at 24 and 48 h. Fungal cultures were reviewed weekly for 4 weeks. Logistic regression analysis with season as a factor and age of the horse as a covariate was performed. Statistical significance was set at P < 0.01. Results Horses ranged from 3 to 24 years of age, with a median age of 9 years. Twenty‐four genera of bacteria and 35 genera of fungi were recovered. Corynebacterium sp., Staphylococcus sp., Bacillus sp. and Moraxella sp. were the bacteria most frequently isolated. Mold species, dematiaceous mold species, Chrysosporium sp., Cladosporium sp., and Aspergillus sp. were the most frequently recovered fungi. Season did not have a significant effect on the presence of microorganisms isolated for individual horses adjusted for age. Younger horses had an increased incidence of gram‐negative rods and fungal isolates. The number of bacteria and fungi isolated are not uniform across seasons. Conclusion There were no significant differences between the number or type of organisms cultured during the sampling seasons in normal Florida horses. A large range of normal bacterial and fungal flora were isolated from these horses. The number of bacteria and fungi isolated are not uniform across seasons. The likelihood of detecting an organism depends on the horses’ age.     

Hydrallantois in two mares

This report describes two mares with hydrallantois. One mare spontaneously entered labor, while the other had labor induced. Uterine inertia necessitated that delivery be assisted in both mares. After delivery, each mare went into hypovolemic shock. One mare survived and the other had to be euthanatized due to rupture of the ventral abdominal musculature with inguinal herniation. The occurrence, diagnosis, treatment and outcome of hydrallantois in the mare are discussed.     

Capture techniques in feral ruminants

This article deals with some of the methods that can be used to capture feral or escaped ruminants. Some of the more common types of live traps, such as snares, spotlighting, and drive nets, are discussed, as are methods of chemical immobilization. The article concludes with a discussion of the medical problems that can occur during capture.     

Uterine culture in mares

A guarded, sterile swab is used to obtain samples for uterine culture. With the mare in stocks, the tail bandage and the perineum washed, the culture rod is introduced into the vagina with a gloved hand. After the rod is guided through the cervix, the guard cap is dislodged and the swab is rubbed along the endometrium, after which the rod is extracted. Samples for uterine culture should only be obtained during full estrus. Swabs should be directly plated onto agar within 2 hours of collection. Blood agar is appropriate for initial screening, but use of specialized types of agar expedites identification of microbes. Plates are incubated at 37 C and inspected for growth every 12 hours. The type and number of bacterial colonies should be coupled with the history and clinical signs in deciding on the necessity and type of treatment. Pure, heavy bacterial growth is usually accompanied by clinical signs of infection. Interpretation of the significance of moderate bacterial growth may be aided by cytologic examination of endometrial smears, made by rolling the swab onto a glass slide and staining with Diff - Quik . Large numbers of neutrophils indicate the need for antibiotic therapy. Mixed bacterial growth and variable numbers of neutrophils usually indicate faulty sampling technic. Microaerophilic or anaerobic cultures may aid diagnosis in cases of equivocal aerobic culture results.     

Embryo transfer in subfertile mares

Four subfertile mares (at least 2 barren years) were used as donors in an embryo transfer program. The embryo recovery rate was 50%, varying between 0-100. The transfer of 9 embryos resulted in the pregnancy rate of 44%. The efficiency of embryo transfer using subfertile mares is low. In this material 20% of flushings resulted in a pregnancy.     

Neospora caninum detected in feral rodents

The role of rodents in the epidemiology of neosporosis was investigated by assaying brain tissue of feral mice (Mus musculus) and rats (Rattus norvegicus) for Neospora caninum. Both mouse and rat brain tissue were extracted for total DNA, and subjected to two different N. caninum-specific nested polymerase chain reaction (PCR) assays. A portion of brain tissue from the mice and rats were also assayed for N. caninum in gerbils or gamma-interferon gene knockout (KO) mice. Of the 105 feral mice tested, 10% were positive in the N. caninum-specific PCR assays. Of the 242 rats tested, 30% were positive in both assays. Although mice and rats had N. caninum by PCR testing, clinical signs of N. caninum infection were not observed nor were N. caninum parasites observed in gerbils or KO mice inoculated with the rodent brain tissue.     

Uterus unicornis in two mares

Two American Paint Horses, a 3-year-old nulliparous mare and a 7-year-old primiparous mare, presented for recent infertility and a pre-breeding examination, respectively. Examination of the internal reproductive tract of both mares using transrectal palpation and ultrasonography revealed the presence of the cervix, uterine body, left uterine horn and bilateral ovaries. The right uterine horn could neither be palpated nor imaged. The clinical diagnosis of uterus unicornis in one mare was confirmed at necropsy, which revealed combined aplasia of the right uterine horn and oviduct.