Nonsurgical recovery of degenerative ova from the uteri of mares.

Degenerated ova were recovered with and without viable embryos 6 days after ovulation in 30 of 210 collections from 24 of 66 mares. All ova were approximately 150 mum in diameter, with an intact zona pellucida and at various stages of cytoplasmic degeneration. Most of the ova were collapsed, although some had an oval appearance. Most of the ova were from maiden 2-year-old mares. Thirty-four of the ova were recovered from the first or second collections. Two ova were recovered from the third collection from two mares. Two degenerative ova per collection were recovered from five collections; three of these collections also contained viable embryos. Degenerated ova were recovered from three mares twice; but not from consecutive collection attempts. Recovered ova were fixed in 2% glutaraldehyde-PBS for scanning electron microscopy. These data indicate that not all unfertilized ova remain permanently in the oviduct; many traverse it and enter the uterus. Furthermore, these data also suggest that when degenerative ova pass into the uterus they either degenerate further and (or) move into the vagina. This is supported by the fact that not all ova can be accounted for when the uterus and (or) the oviducts are washed.

Digestion of hay protein in different segments of the equine digestive tract.

Mature ponies fitted with permanent ileal cannulas were used in a 3 X 3 Latin square experiment to quantify prececal, postileal and total tract digestion of hay protein. Coastal Bermuda grass (CB), containing 11.7% crude protein, and two qualities of alfalfa, containing 15.0% (low-protein alfalfa; LA) and 18.1% (high-protein alfalfa; HA) crude protein, were fed in coarsely chopped form at 2% of body weight daily. Total tract apparent digestibility of the N in HA (73.8%) was higher than that in CB (57.0%; P less than .05) and was slightly higher than that in LA (66.1%; P less than .10). Nitrogen in LA was apparently more digestible than that in CB (P less than .05). Apparent prececal digestibilities of N in LA and CB were 1.3% and 9.6%, respectively, and were lower (P less than .05), or tended to be lower (P less than .10), than the 21.0% observed for HA. In relative terms, an average of 9.4% of the total N digestion occurred in the upper tract when CB and LA were fed, whereas 28.5% of total N digestion occurred in the foregut when HA was fed. There was a slightly higher concentration of total plasma free amino acids (P less than .10) at 1 h postfeeding when horses were fed alfalfa. Also, N retention was higher when ponies were fed HA (P less than .05) than when LA or CB were fed. Apparent postileal N digestibility was 52.5% for CB, 65.7% for LA and 66.9% for HA. Differences were not significant, and the large intestine appeared to compensate for the inefficiency of N digestion in the upper tract.(ABSTRACT TRUNCATED AT 250 WORDS)

Nitrogen metabolism in mature horses at varying levels of work.

Nitrogen metabolism was measured in five mature geldings performing varying levels of work that simulated race training. Following an initial maintenance period without forced exercise, workload was increased in succeeding 18-d periods by doubling the distance the horses were galloped in each period from period 2 through 4. A 4-d N balance experiment was conducted at the end of each 18-d period. The maximum distance galloped daily, which was in period 4, was repeated in period 5. Then, workloads were decreased through period 7. Nitrogen balance was determined in two additional periods, 8 and 9, while the geldings were at maintenance with no forced exercise. Geldings were fed diets containing similar protein-to-calorie ratios in all periods. Nitrogen retention increased (P less than .05) as workload increased from period 1 to period 3 and remained elevated throughout the rest of the experiment. Nitrogen retention did not (P greater than .05) decrease from highest values, observed at maximum workload during periods 4 and 5, until period 9, which was 34 d postexercise during a maintenance period. Nitrogen retention was higher (P less than .05) following training than before training.

Methods for collecting follicular oocytes from mares.

A series of experiments was conducted to develop a procedure for consistent, repeatable collection of oocytes from the preovulatory follicle of the mare. In one experiment, in situ follicular aspiration with a needle and syringe was performed on 19 mares. From 37 aspirations, four oocytes were recovered (10% recovery rate). In a second experiment, ovaries were visualized via standing flank laparotomy during which two different aspiration techniques were used. Use of a needle and syringe as in the first experiment resulted in successful oocyte recovery in one of seven (14%) attempts. Aspiration via a continuous irrigation vacuum system (CIV), developed for use during laparotomy, resulted in collection of oocytes from six of 10 (60%) attempts. In the third experiment, oocytes were recovered from seven of 18 (38%) attempts at in situ follicular aspiration using a double-lumen needle attached to the CIV. In each experiment, some mares were subjected to stimulation of follicular maturation by exogenous hormones. Oocyte recovery was significantly increased in treated mares as compared with nontreated mares. Results indicate that collection of equine follicular oocytes by in situ aspiration is possible with moderate success. Oocytes apparently are not physically damaged by the procedure, as most retained either the corona radiata or the entire cumulus cell mass.

Use of different nonglycolysable sugars to maintain stallion sperm viability when frozen or stored at 37 degrees C and 5 degrees C in a bovine serum albumin medium.

Bovine serum albumin (BSA) diluents containing lactose, raffinose or sucrose were not different (P greater than 0.05) in their ability to maintain stallion sperm viability, as determined by percentage motile spermatozoa (PMS) and their rate of forward movement (RFM), when stored at 37 or 5 degrees C for 24 h. These diluents did promote a higher (P greater than 0.05) PMS and RFM, when compared with BSA diluents containing arabinose or galactose. The BSA-arabinose and BSA-galactose diluents did not differ (P less than 0.05) in their ability to support sperm viability and were detrimental to spermatozoa. The fertility of freshly collected and diluted spermatozoa was not different (P greater than 0.05) when extended in BSA diluents containing lactose, raffinose or sucrose. There was no difference (P greater than 0.05) in PMS and RFM of frozen-thawed stallion spermatozoa when the spermatozoa were frozen, thawed and incubated at 37 degrees C for 180 min in BSA diluents containing lactose, raffinose or sucrose. Spermatozoa from 6 of 8 stallions did not survive the freezing process. A one-cycle conception rate of 32% was obtained from frozen-thawed spermatozoa extended in BSA diluents containing lactose, raffinose or sucrose. This rate was 78% of the conception rate obtained when the same mares were inseminated with fresh semen in a subsequent study (41%).

Motility and fertility of equine spermatozoa extended in bovine serum albumin and sucrose.

Inclusion of either 1 or 3% (w/v) bovine serum albumin (BSA) in 8.6, 10, or 12% sucrose enhanced the maintenance of equine sperm motility in vitro at 38 degrees C for 8 h. There was a trend toward higher percent motile spermatozoa (PMS) at 16 and 24 h of incubation in semen samples containing BSA than in those that did not. The highest concentration of sucrose (12%) was slightly less effective in supporting PMS than either of the lower concentrations. However, sucrose concentrations had no apparent effect on rate of forward movement (RFM) of spermatozoa. Pregnancy and foaling rates were similar for mares inseminated with semen extended in either cream gel or 3% BSA-10% sucrose.

Inclusion of bovine serum albumin in semen extenders to enhance maintenance of stallion sperm viability.

Semen from seven mature stallions was used to test the motility response of sperm cells when 3% bovine serum albumin (BSA) was added to seminal plasma and skim milk diluents. A total of 45 ejaculates was collected by artificial vagina and immediately evaluated for percent motile spermatozoa (PMS), rate of forward movement (RFM) and sperm cell concentration. Aliquots (four from each ejaculate) of raw semen containing 500x10(6) sperm cells were exposed to each of the following treatments: (1) seminal plasma (SP), (2) SP+BSA, (3) skim milk (SKM), (4) SKM+BSA; and incubated in 50-ml tubes at 37 C. The sperm cell characteristics, PMS and RFM, of each treatment suspension were reevaluated at 0, 0.5, 1, 2, 6, 12, 18 and 24 hr post-treatment. Inclusion of BSA and the type of extender, either seminal plasma or skim milk, significantly (P<0.05) affected the PMS and RFM of spermatozoa. Analysis of means within evaluation times showed that PMS maintenance was enhanced (P<0.05) when BSA was included in extenders at all incubation intervals except 24 hr. SKM+BSA maintained the highest (P<0.05) PMS for the first 2 hr with SP+BSA sustaining the highest (P<0.05) PMS from 12 to 24 hr. Skim milk alone sustained higher (P<0.05) PMS than the SP diluent for the first 6 hr of incubation, whereas SP maintained a higher (P<0.05) PMS than SKM from 18 to 24 hr. The RFM of spermatozoa was greatest (P<0.05) for the first 6 hr of incubation when exposed to SKM+BSA. Seminal plasma + BSA sustained a higher (P<0.05) RFM for the first 6 hr of incubation than SP alone, but not higher than SKM at this interval. Skim milk sustained a higher (P<0.05) RFM of spermatozoa for the first 6 hr of incubation than SP. These data support the hypothesis that BSA protects spermatozoa from the harmful effects of lipid peroxidation. Including this substance in semen extenders may prolong maintenance of sperm motility.

Influence of photoperiod on the pregnant and postpartum mare.

Over a 2-year period, mares (n = 32) of predominantly Quarter Horse breeding and their foals were used to study the effects of photoperiod on reproduction during the periparturient period. Specific objectives of the study were (i) to evaluate the effects of an extended photoperiod regimen (16 hours' light) on gestation length, foal development, and postpartum reproductive performance, and (ii) to measure changes in plasma progesterone concentrations during the last trimester of gestation and plasma luteinizing hormone (LH) concentrations after foaling until ovulation. Mares in the extended daylength treatment group had shorter (10 days) mean gestation length (P less than 0.01) than did control mares (exposed to normal daylight hours). Although foals of mares exposed to long photoperiod were carried for a shorter term, their birth weights tended to be heavier, but not significantly, than those of foals from control mares. Foal size, as determined by body measurements, was not affected by the photoperiod. Neither interval from parturition to 1st ovulation nor onset of estrus to ovulation were significantly affected by the extended photoperiod. Plasma progestogen concentrations increased from basal levels at prepartum weeks 4 and 5 to peak values a week before foaling in both mare groups. The LH profiles from parturition to ovulation were similar for both mare groups. However, mean plasma LH values during the 7 days before the 1st ovulation were higher (P less than 0.05) in mares exposed to extended daylength than in control mares.

Induction of parturition in the mare with prostaglandin F2 alpha.

Thirty-one mares of Quarter Horse and Thoroughbred breeding were utilized in two experiments to evaluate the efficacy of prostaglandin F2 alpha (PGF2 alpha) for induction of equine parturition and to monitor the effects of this treatment on viability of the resulting foals. Three of five mares given 5 mg PGF2 alpha (im) on day 338 of gestation foaled 19.6+/-8.2 hr postinjection. In the second experiment immediately following 3 daily injections of 10 mg estradiol cypionate (ECP) given on days 326, 327 and 328 of gestation, seven mares were infused (iv) with PGF2 alpha at the rate of 1.3 mg/hr for 24 hr or until parturition occurred. Four of the seven mares foaled in 8.8+/-1.8 hr after the start of infusion. Side effects including sweating, hypothermia, increased respiration rate and diarrhea were evident in both injected and infused mares, but effects were transient. Neither the injection, nor infusion route of administration of PGF2 alpha adversely affected the viability of foals. However, some mares induced to foal 12 days prior to expected parturition had foals with slightly weaker pasterns than those of control mares.

Influence of prostaglandin F2 alpha on sperm production and seminal characteristics of the stallion.

Six mature stallions were used to test the effect of prostaglandin F2 alpha (PGF2 alpha ) on sperm production and seminal characteristics. Semen was collected from each stallion twice weekly 1 hr following a 10 mg intramuscular injection of PGF2 alpha or a sham injection. A switchback design was used so that three stallions received PGF2 alpha and three served as controls during the first 9 weeks (period 1). Treatment regimens were reversed during the second 9 weeks (period 2). Treatment of stallions with PGF2 alpha resulted in an increase (P less than .05) in gel free seminal volume and a decrease in sperm cell concentration. Total spermatozoa, sperm cell motility, and percentage of primary and secondary sperm abnormalities of ejaculates were not significantly affected by treatment of stallions with PGF2 alpha before semen collection. All treated stallions exhibited a pronounced sweating response to the drug. During the experiment, two of the six stallions masturbated within 20 to 30 minutes after PGF2 alpha treatment without achieving an erection.

Effect of calcium and fatty acids on the isolation of stallion spermatozoa in BSA.

Fifty ejaculates, ten from each of 5 mature stallions, were utilized to study the effects of calcium and fatty acids on equine spermatozoa which were isolated in 3% Bovine Serum Albumin (BSA). The ejaculates were evaluated for percent motile spermatozoa, rate of forward movement, debris, primary abnormalities and secondary abnormalities. The isolation procedure consisted of layering 2 ml of diluted semen (100 x 10(6) spermatozoa/ml) over 6 ml of 3% BSA in 13 x 125 mm columns in a water bath (37 degrees C). After 30 min., the top semen layer and upper half of the BSA layer were withdrawn from all columns and the lower half of the BSA was re-evaluated. A 2 x 2 factorial arrangement of treatments was utilized with either the inclusion or omission of calcium or fatty acids in the BSA isolation media. The percent motile spermatozoa and rate of forward movement were increased (P<.01) when fatty acids were included in the isolation media but decreased (P<.01) when they were omitted. The highest percent motile spermatozoa and rate of forward movement were observed with BSA in the presence of fatty acids and omission of calcium. The calcium by fatty acid interaction, stallion effect and stallion by treatment interaction were significant for percent motile spermatozoa. Less debris was observed in all samples of isolated spermatozoa when compared with the initial estimate. Isolation resulted in a reduction of (P<.01) the primary abnormalities. Also, fewer (P<.01) secondary abnormalities were observed after isolation in all treatments except 4 (-FA+Ca) than were found in the ejaculate sample.

Effects of restrictive suckling on postpartum reproductive performance in mares.

Quarter Horse mares (n = 30) and their foals were used in a 2-year study. Objectives of the study were (i) to determine the effects of a treatment regimen, which permitted nursing for four 1-hour periods each day, on the postpartum reproductive performance of the dams, and (ii) to assess the effects of this procedure on the growth and development of foals. Mares in the restricted suckling treatment group showed estrus and ovulated sooner (P less than 0.01) after parturition than did control mares during the first year of the study, but not during the second year. Plasma progesterone concentrations were similar for both treated and control mares and remained less than 1 ng/ml from parturition to ovulation. Conception rates were not significantly influenced by treatment. However, two mares which had records of failing to have estrus while nursing a foal were bred and ovulated, and one of the two conceived while on the restricted suckling treatment. The absence of treatment effects on any of the variables measured in the foals indicates that restricted suckling did not impair foal growth and development.