James G. W. Wenzel

Pluronic® F127 gel formulations of deslorelin and GnRH reduce drug degradation and sustain drug release and effect in cattle

The objective of this study was to compare the effectiveness of intramuscular sustained release Pluronic F127 (PF127) gel formulations of deslorelin, a potent GnRH agonist, and GnRH to their solution formulations in inducing the release of luteinizing hormone and formation of luteal tissue in cattle. Injectable gel formulations of deslorelin and GnRH were prepared using Pluronic F127 (25%, w/w), a block copolymer. PF127 gels sustained the in vitro release of deslorelin as well as GnRH at similar rates and reduced drug degradation in muscle tissue when compared to the solution formulations. Deslorelin, as well as GnRH, elicited desirable elevations in plasma LH and progesterone concentrations in vivo. When compared to the solution formulations, the gel formulations of both drugs induced a broader peak of LH. Also, the peak LH levels were lower and the peak times were delayed with the gel formulations compared to the solution formulations. While the solution dosage form of deslorelin and GnRH elicited similar responses, the PF127 gel formulation of deslorelin induced peak LH levels at an earlier time (3 h for deslorelin versus 5.25 h for GnRH). The results indicate that, deslorelin exerts a pharmacological effect in cattle. The LH response to deslorelin as well as GnRH can be altered by controlling the input or the release rate of the drug. PF127 gel formulations can sustain peptide release and reduce peptide degradation.

Adaptation of the hypoosmotic swelling test to assess functional integrity of stallion spermatozoal plasma membranes.

Hypoosmotic swelling (HOS) is used for assessing plasma membrane function and fertilizing capacity of human spermatozoa. However, HOS solutions and methodologies have not been evaluated specifically for assessing stallion spermatozoa. The objective of this study was to identify a HOS solution and assay conditions specifically for stallions that would maximize spermatozoal plasma membrane swelling. The HOS solutions and assay conditions, including incubation time (15 to 180 min), temperature (25 degrees vs 37 degrees C), and total number of cells examined (100, 200 or 500) were evaluated. Assay consistency, accuracy, reliability and repeatability also were determined. Maximum spermatozoal plasma membrane swelling was observed in a 100 mosmol sucrose solution (P < 0.001). Incubation time (P = 0.67), temperature (P = 0.70) and total number of spermatozoa examined (P = 0.38 and P = 0.24 for 100 vs 200 and 100 vs 500, respectively) did not influence percent of HOS positive spermatozoa observed. A high degree of assay accuracy was indicated when a correlation of r = 0.998 was obtained between the HOS positive spermatozoa observed and expected when known amounts of heat-treated spermatozoa, unable to undergo swelling, were added to untreated spermatozoa. Assay consistency was demonstrated, as reflected by a mean coefficient of variation of 0.073 for 4 stallions. Also, the coefficient of variation from 2 analyses of variance was 0.168 and 0.096, indicating reasonably good assay reliability; estimates of repeatability from the same analyses were 0.794 and 0.968. The HOS test adapted to stallion spermatozoa in this study is a simple, highly accurate and consistent assay with good reliability and repeatability. Results observed under the conditions evaluated also permit some flexibility in adapting this assay to individual laboratory and practice settings for evaluating stallion spermatozoal plasma membranes.

Effect of administering oxytocin or cloprostenol in the periovulatory period on pregnancy outcome and luteal function in mares.

Mares (n = 37) were treated from 4h after breeding through 2 days post-ovulation with oxytocin or cloprostenol. Oxytocin (20 units i.m.) was administered every 6 h and cloprostenol (250 mcg i.m.) daily. Luteal function was impaired for several days following treatment, however, lower progesterone levels among cloprostenol treated mares in this study did not result in decreased pregnancies. Pregnancy outcome at 15 days post-ovulation was not different between the oxytocin (13/18) and cloprostenol (13/19) treatment groups, respectively (P = 0.80). The results of this study indicate cloprostenol can be used to treat post-breeding mares through the second day following ovulation without decreasing pregnancy outcome.

Observations using the new bull-breeding soundness evaluation forms in adult and young bulls.

The new bull-breeding soundness forms were used to evaluate 1276 bulls; 62.85% of these bulls were deemed satisfactory. Unsatisfactory and deferred percentages were 28.92% and 8.23% respectively. Reasons for the various classifications were evaluated, and age and breed comparisons were made.

Pregnancy outcome in mares following insemination deep in the uterine horn with low numbers of sperm selected by glass wool/Sephadex filtration, Percoll separation or absolute number

Mares were inseminated deep in the uterine horn with 25 million sperm selected by glass wool/Sephadex (GWS) filtration, Percoll separation (PS) or absolute number (AN). Deep-horn insemination using a low-volume, smooth tipped, flexible pipette/catheter delivery system allowed more efficient use of stallion sperm and reduced post-breeding uterine reaction in mares. Mares were pregnant in 15/30, 13/30 and 10/30 cycles for GWS, PS and AN selection methods, respectively. Sperm selection method did not effect pregnancy outcome (P=0.422). However, sperm selected for deep-horn insemination by filtration through a glass wool/Sephadex column tended to improve fertility over simply using an absolute number of sperm (P=0.105).

Retrospective analysis of the management of 78 cases of postpartum metritis in the cow

Case records of post partum metritis in cows were obtained from Auburn University (42 cases) and Tuskegee University (36 cases) Large Animal Hospital files. Dairy cows (n=68) comprised 87.2% of the cases and beef (n=10) 12.8%. Age did not differ between the types, but the number of postpartum days at admission (mean+/-1 SD) differed significantly (P=0.02) between dairy (7.3+/-4.27) and beef (3.7+/-4.06) cows. Older cows and those with hyperthermia were less likely to recover from puerperal metritis (P=0.07 and 0.03, respectively). Type of treatment had no significant association with survival. The number of days of hospitalization was associated with institution (P<0.001), number of postpartum days at admission (P=0.001), history of predisposing factors (P=0.0006), hyperthermia (P=0.006), and length of various treatments (P<0.001). However, the type of treatment had no significant association with length of hospitalization. This study suggests that the necessity for or type of therapy has little effect on the outcome of postpartum metritis.

Luteal function in mares following administration of oxytocin, cloprostenol or saline on day 0, 1 or 2 post-ovulation.

Mares (n = 30) were treated in the post-ovulatory period with saline, oxytocin, or cloprostenol (Clo). Dose, administration frequency and treatment day (Day 0, 1 or 2 post-ovulation) were evaluated. Interovulatory interval of control cycles was 22.7 (+/-0.36) days with a range of 20.6 (+/-1.44) to 23.8 (+/-1.39) days among all treatment groups. Mares treated with two micro-doses of cloprostenol on Day 2 post-ovulation had the shortest interovulatory interval. This group also had the lowest mean circulating progesterone concentrations on Days 3-7 and 13, and was the slowest group to reach concentrations of 5 ng/ml. Repeated administration of cloprostenol over 24 h in the early post-ovulatory period may more effectively impair luteal function than single doses. This could negatively affect pregnancy outcome but may be effective for lysing the early post-ovulatory luteal structure when mares are not bred.

Effects of grazing endophyte-infected tall fescue on eCG and progestogen concentrations from gestation days 21 to 300 in the mare

Abstract The influence of grazing endophyte-infected tall fescue on endometrial cup formation and function, progestogen production, and embryonic and fetal development were examined in pregnant mares between Day 21 and Day 300 of gestation. Total immunoreactive progestogens and equine chorionic gonadotropin (eCG) concentrations were compared between untreated controls (endophyte-free, n = 12) and treated mares (endophyte-infected, n = 12). There were no differences in endometrial cup formation or function, as determined by eCG concentrations at Days 45, 60, 75, 90 and 120 (P > 0.05) between mares grazing endophyte-infected and endophyte-free tall fescue. Mares grazing the endophyte-infected tall fescue had lower total progestogen concentrations (P

A comparison of physical examination and clinicopathologic parameters between sheared and nonsheared alpacas (Lama pacos)

The purpose of this study was to determine the physiological changes associated with chronic heat stress in sheared versus nonsheared alpacas. Fourteen intact male adult alpacas were randomly assigned to one of the two groups: Group S alpacas were sheared to within 2cm of their skin; Group NS alpacas were not sheared. These animals were maintained from June through August in east central Alabama. Data collected in the morning, every two weeks, included vital signs, body weight, body condition score, complete blood counts, serum chemistries and electrolytes, whole blood selenium, and plasma cortisol. S and NS groups were contrasted using the repeated measures analysis of variance, and pertinent correlations with weather parameters were calculated. Clinical heat stress was not evident in any animals during the study. Significant differences between treatment groups were seen in rectal temperature (P=0.0095), sodium concentration (P=0.0219), and blood urea nitrogen (BUN) (P=0.0189). The mean rectal temperature of the NS group was above the normal range on five sampling times compared to only once for the S group. However, mean sodium and serum urea nitrogen levels were within normal limits in both groups at all sampling times. Rectal temperature of only the S group was positively correlated to weather parameters. Sodium of both S and NS groups and BUN of the NS group were negatively correlated with weather parameters. This study indicates that there are differences between sheared and nonsheared alpacas in physical examination and clinicopathologic parameters that can be correlated with changes in ambient conditions. These differences suggest that nonsheared alpacas are less heat tolerant than sheared alpacas. Therefore, shearing is recommended for animals exposed to similar conditions.

Histological and Histochemical Characterization of the Bovine Rete Ovarii through the Estrous Cycle and Gestation

The rete ovarii of cattle may be morphologically divided into the intraovarian rete, the connecting rete, and the extraovarian rete. The connecting rete may be subdivided into that portion which lies within the ovary proper and that portion that lies in the mesovarium. The portion of the connecting rete which lies in the mesovarium is connected to the infundibulum of the uterine tube by a patent tubo‐retial communication.