Efficacy of benzimidazole anthelmintics in goats and sheep in the Philippines using a larval development assay.

The negative effects of nematodes in small ruminants can be reduced by use of dewormers but their effectiveness is increasingly limited by the emergence of anthelmintic resistance. The efficacy of benzimidazole (BZ) anthelmintics in the Philippines was estimated by an in vitro larval development assay using worm eggs recovered from faeces collected from goats and sheep. Two hundred and eighteen farms were selected to represent areas of the country with high goat and sheep populations and the full range of farm sizes, from smallholders with just a few animals to commercial and institutional farms with several hundred. Initial surveys of worm control advisers indicated that BZs have been in continuous widespread use for up to 20 years with little use of other chemical groups. A larval development assay (LDA: DrenchRite) was modified for use with BZs alone to allow up to five samples to be analysed on a single microtitre plate. The assay was validated by comparison with the faecal egg count reduction test (FECRT). The dominant nematode genera were Haemonchus and Trichostrongylus with small numbers of Oesophagostomum. The range of BZ efficacy estimated from the LDA results was 0-100% and the distribution of efficacy levels was continuous, with mean efficacy of 82 and 64% for goats and sheep, respectively. There were significant associations between efficacy and parameters measured to characterize the sampled farms: size of animal management group, FEC of sample, recent importation of stock and no access to common grazing were all correlated with decreased efficacy. Likewise, low efficacy was associated with reported frequency and number of years that BZ drenches had been used. The LDA was found to be highly suited to estimate efficacy in nematode populations from small farms where performance of a FECRT for even one chemical would be impractical. Using a larval development assay, we have demonstrated a wide efficacy range for BZs against nematodes from all sizes of goat and sheep farms in the tropics.

Local distributions of oviductal estradiol, progesterone, prostaglandins, oxytocin and endothelin-1 in the cyclic cow.

The cyclic patterns of hormones which regulate the activity of the oviduct in the cow have not been adequately reported. We studied progesterone (P4), estradiol 17 beta (E2), prostaglandin E2 (PGE2), prostaglandin F2 alpha (PGF2 alpha), oxytocin (OT) and endothelin-1 (ET-1) concentrations in the cow oviduct. Reproductive tracts from cyclic Holstein cows in the follicular phase (n = 5), post ovulation phase (n = 5) and luteal phase (n = 5) were collected at a slaughterhouse. Oviducts were separated from the uterus, the lumen vas washed with physiological saline, and the enveloping connective tissues were removed. The fimbria was then separated at first and then the rest was divided into 2 parts of equal length (proximal and distal). After extraction, levels of different hormones in the tissues were measured using double antibody enzyme immunoassays (EIAs). There were no differences in any hormone concentration between the 3 parts of the oviduct at any stage of the estrous cycle. The highest concentration of oviductal P4 was observed during the luteal phase and in the oviduct ipsilateral to the functioning CL. Oviductal OT was unchanged throughout the cycle. The highest E2 concentration was observed during the follicular phase in the oviduct ipsilateral to the dominant follicle. The oviduct ipsilateral to the dominant follicle during the follicular phase and ipsilateral to the ovulation site post ovulation showed higher levels of PGE2, PGF2 alpha and ET-1 than those on the contralateral side or during the luteal phase. The highest PGE2 was observed in the oviduct ipsilateral to the ovulation site during the post ovulation phase. The results suggest that the ovarian products (P4, OT and E2) and the local oviductal products (PGE2, PGF2 alpha, and ET-1) may synergistically control oviductal contraction for optimal embryo transport during the periovulatory period, and provide further evidence for the local delivery of ovarian steroids to the adjacent reproductive tract.

The effect of viscosity of semen diluents on motility of bull spermatozoa.

The effect of egg yolk extender on semen viscosity and bull sperm motility of fresh and cooled or deep frozen semen was determined by a computer-assisted system. Viscosity of the extender was determined by flow time. Based on the sperm velocity (velocity of the average path), individual spermatozoon were classified into groups of progressively motile (>==30 microm/sec) and immotile (<10 microm/sec) spermatozoa. The average velocity of progressively motile spermatozoa (VPM), the velocity of linear progressively motile spermatozoa (VLP) and the percentage of linear swimming spermatozoa (LIN) were evaluated. The addition of 10, 20 or 30% egg yolk to Tris buffer (pH 6.5) resulted in a linear decrease of VPM and a decrease in the percentage of progressively motile spermatozoa, but it increased the relative rate of LIN in fresh diluted semen. Increasing the levels of egg yolk in the diluent resulted in higher viscosity. The VLP was significantly higher than the VPM. In refrigerated or frozen semen samples, extender with 30 and 20% egg yolk had a similar effect on the VPM but not on the percentage of progressively motile sperm cells. Freezing of egg yolk (30%) extender to -20 degrees C resulted in a significant increased flow time and higher viscosity. Dilution of semen samples with high viscosity extender decreased the VPM in fresh and chilled semen. Freezing semen of high viscosity extender with glycerol had no apparent effect on the percentage of progressively motile spermatozoa compared with that of non-glycerinated egg yolk extender. The results suggest that different concentrations of egg yolk in the extender can influence the parameters of semen viscosity and sperm motility evaluated by a computer-assisted system.

Oviductal progesterone concentration and its spatial distribution in cyclic and early pregnant cows.

Changes and local distribution of oviductal progesterone (P(4)) concentration during the estrous cycle and early pregnancy in cows were investigated. Intact reproductive tracts were collected from 16 Holstein cows at an abattoir. Samples were classified in to 4 stages (follicular, postovulatory, luteal and early pregnant,< 20 d) based on visual observation of corpus luteum (CL), uterine characteristics and luteal P(4) concentrations. Oviducts were separated from the uterus at the utero-tubal junction and divided into 4 parts: fimbriae, proximal, medial and distal parts. Luteal tissue samples were also collected. Progesterone levels in oviductal and luteal tissues were determined by radioimmunoassay (RIA). Comparatively higher (P < 0.001) P(4) levels were found in stages with a functioning CL ( luteal phase and early pregnancy) than in those with a regressing CL (follicular phase and post ovulation). The oviduct ipsilateral to the CL bearing ovary during the luteal phase and early pregnancy showed higher ( P < 0.001) P(4) concentrations than the contralateral side. Such a difference was not observed during the follicular phase or post ovulation. The ipsilateral oviduct to the functioning CL at early pregnancy showed higher (P <0.05) P(4) levels than at the luteal phase, while no significant difference in luteal P(4) levels between these 2 stages was observed. Neither were any differences in P(4) concentration within the oviduct observed during any phase of the estrous cycle or during early pregnancy. A positive relationship between luteal and oviductal P(4) concentrations was noted. In conclusion, changes in P(4) levels in the oviduct depend on the location and functional stage of the CL. Localized levels of P(4) in the oviduct may be due to local delivery of P(4) from the CL.

Evidence of ovulation in goats (Capra hircus ) with short estrous cycle and its occurrence in the tropics.

The influence of season and exposure to buck on the length of the estrous cycle were determined in 32 normally cycling native goats of different ages and parity. The estrous cycle durations were classified as short, medium and long. Ten goats (31%) exhibited medium length estrous cycles; 22 animals (69%) exhibited a combination of short, medium and long estrous cycles. Of the 155 estrous cycles studied, 15% were short, 72% were medium and 13% were long. Short estrous cycles were significantly shorter (P < 0.01) in estrus duration than medium or long estrous cycles. Significant differences (P < 0.05) for estrous cycle length and estrus duration were found between does but not within does. The presence of a buck for 8 to 16 h showed no significant effect on the length of the estrous cycle or on estrus duration, but a 24-h exposure period to a buck yielded shorter estrous cycles and estrus duration than found in the unexposed control group. Estrous cycles were significantly shorter (P < 0.01) in October, when rainfall is 57.9% of the annual total, and significantly longer (P < 0.01) in February, when rainfall is low at 0.2% of the annual total. A negative linear relationship (r = -0.87) was observed between estrous cycle and rainfall. Twenty-eight (90.3 %) of the short estrous cycles were ovulatory. The short cycles had a relatively lower ovulation rate than medium and long estrous cycles. The results indicate that the short estrous cycle in goats is associated with ovulation, and its occurrence in the tropics is related to rainfall.

Prostaglandin E2 levels in uterine tissues and its relationship with uterine and luteal progesterone during the estrous cycle in dairy cows.

Endometrial tissue homogenates obtained at luteal and follicular stages of the estrous cycle were determined for prostaglandin E2 and progesterone contents by EIA and RIA, respectively. In Experiment 1, the concentrations and changes of PGE2 in uterine tissues collected by biopsy before slaughter and subsequent samples collected at 30, 60 and 90 min after slaughter were measured. No significant differences were observed in the concentration of PGE2 preslaughter or at 30 and 60 min post slaughter. However, there was a significant decrease (P<0.01) in PGE2 concentration 90 min post slaughter. In Experiment 2, the concentrations of PGE2 in the ipsilateral and contralateral horns in relation to corpus luteum function were compared. A significant (P<0.05) interaction was found between stages of estrous cycle (luteal vs follicular) based on CL progesterone content, and type of uterine horn (ipsilateral vs contralateral) on uterine PGE2 levels. The PGE2 concentration was significantly higher (P<0.01) at luteal phase than at follicular phase. During the luteal phase PGE2 concentrations in tissues of the uterine horn ipsilateral to the corpus luteum was significantly higher (P<0.01) than the contralateral horn. The PGE2 concentration was low and did not differ significantly between horns during follicular phase. A parallel increase (luteal: high) and decrease (follicular: low) in PGE2 and progesterone concentrations were observed. Correlations were observed for CL progesterone and uterine PGE2 concentrations as well as for PGE2 and progesterone concentrations in uterine tissues (r=0.70 and r=0.60, respectively). The results show that the increase in PGE2 concentrations in uterine tissues coincides with the high uterine progesterone concentrations during luteal phase.

Serum levels of testosterone and oestrogens in normal and infertile stallions.

The serum concentrations of testosterone and oestrogens were determined in stallions classified as geldings, normal (according to age) or infertile (azoospermic). There were significant differences in testosterone and oestrogen levels between the groups. Normal concentrations of testosterone and total oestrogens were attained after 16 months of age. Castrated and immature horses (< 15 months of age) had lower levels of testosterone and total oestrogens than did mature and infertile stallions. Azoospermic stallions had lower levels of testosterone and total oestrogens than did normal mature horses except in one case (B4) in which the level of total oestrogens was increased markedly. Critical ellipses for testosterone and total oestrogens were obtained for both groups (6-15-month-old colts and stallions aged > 4 years). The results suggest that serum levels of testosterone and total oestrogens may provide a sensitive index of the endocrine function of the testis in male horses.