A comparison of diagnostic techniques for postpartum endometritis in dairy cattle.

Holstein cows (n=221) from eight commercial dairy herds were examined for endometritis between 28 and 41 days postpartum using 5 diagnostic techniques: (1) vaginoscopy; (2) ultrasonographic assessment of uterine fluid volume; (3) ultrasonographic assessment of endometrial thickness; (4) endometrial cytology collected by cytobrush; and (5) endometrial cytology collected by uterine lavage. Concordance correlation was used to evaluate the reliability of cytobrush and lavage cytology. Cytobrush cytology was found to have the greatest intraobserver repeatability (cytobrush, rho(c)=0.85 versus lavage, rho(c)=0.76) and was chosen as the reference diagnostic test. Pregnancy data at 150 days postpartum was available for 189 cows. Survival analysis was used to determine the lowest percentage of polymorphonuclear cells associated with time to pregnancy. The sensitivity and specificity of the diagnostic techniques was determined using pregnancy status at 150 days and cytobrush cytology as the diagnostic standards. The risk of non-pregnancy at 150 days was 1.9 times higher in cows with more than 8% PMNs identified using cytobrush cytology than in cows with less than 8% PMNs (P=0.04). Twenty-one cows of 189 cows (11.1%) had >8% PMNs and were considered to be positive for endometritis. Cows with endometritis had a 17.9% lower first service conception rate (P=0.03) and a 24-day increase in median days open (P=0.04). The sensitivities of all five diagnostic tests relative to 150-day pregnancy status ranged from 7.1 to 14.3% and the specificities from 84.0 to 93.3%. Relative to cytobrush cytology, the respective sensitivity and specificity values are as follows: vaginoscopy (53.9%, 95.4%); lavage cytology (92.3%, 93.9%); ultrasonographic assessment of uterine fluid (30.8%, 92.8%); and ultrasonographic assessment of endometrial thickness (3.9%, 89.2%). Endometritis impaired reproductive performance. Cytobrush cytology was the most reliable method of diagnosing endometritis in cattle.

Ovarian follicular development and endocrine responses in follicular-fluid-treated and hemi-ovariectomized heifers.

The effects of charcoal-extracted bovine follicular fluid (BFF) on endocrine profiles and follicular development in intact and hemiovariectomized postpubertal heifers were examined. Oestrus-synchronized heifers received Norgestomet implants on day 1 and 7 of treatment and were then injected s.c. with 11 ml saline (control) or 11 ml BFF twice a day for 12 days. The ovary bearing the largest follicle (OV1) was removed on day 7 and the remaining ovary (OV2) was collected on day 13. Follicles were observed by daily ultrasonography and were classified according to diameter (size 1: 2-3 mm; size 2: 4-6 mm; size 3: 7-10 mm; size 4: > 10 mm). After ovariectomy they were classified by diameter and histologically as normal or atretic. Intact control heifers had increased numbers of size 4 follicles on OV1 on days 6 and 7; no increase was observed in BFF-treated heifers (P < 0.03). In BFF-treated heifers, the mean basal LH concentration was higher (P < 0.05) and that of FSH was lower (P < 0.04) than in controls. FSH concentrations in BFF-treated heifers decreased from 0.60 +/- 0.08 ng ml-1 (day 1) to 0.22 +/- 0.05 ng ml-1 (day 7; P < 0.04). The concentration of oestradiol increased in control heifers, but not in BFF-treated heifers (P < 0.001). After hemicastration, OV2 underwent compensatory hypertrophy in control heifers, with an increase in the number of size 2, 3 and 4 follicles (P < 0.05), whereas BFF-treated heifers did not. Thus, total follicular volume was much lower in BFF-treated than in control heifers on day 13 (92.2 +/- 15.4 versus 1393.8 +/- 276.6 mm3; P < 0.0002). A transient increase in FSH (P < 0.006) and oestradiol (P < 0.01) concentrations occurred after hemiovariectomy in control but not in BFF-treated animals. In control heifers, an analysis of temporal relationships showed negative correlations between the volume of size 3 and size 4 follicles, and between FSH concentrations and the volume of size 3 and 4 follicles. A positive correlation was found between the mean diameter of the largest follicle and the concentration of oestradiol, whereas negative relationships were found between the concentrations of FSH and oestradiol, and between FSH and the mean diameter of the largest follicle. Analysis of the histological data showed that the number and volume of follicles > 8.57 mm was lower in the BFF-treated OV1 ovary, whereas no differences were found for follicles < or = 8.57 mm.(ABSTRACT TRUNCATED AT 250 WORDS)

A survey of veterinarian and producer perceptions of herd health services in the Saskatoon milkshed.

A cross-sectional survey was conducted to determine the availability, rates of adoption, and producer perceptions of veterinary services in five areas of dairy production medicine: reproduction, milk quality, nutritional consulting, infectious disease control, and heifer rearing. Questionnaires were completed by all veterinary clinics and 86% of the dairy producers in the Saskatoon milkshed. Veterinary perceptions of services offered were compared with farmer perceptions of services received. The veterinary clinics appeared to overestimate their services in the areas of nutritional consulting and heifer rearing. The primary determinant of a producer's perception of being on a herd health program was the occurrence of regularly scheduled reproduction visits. Producers who perceived themselves as being on a herd health program also believed that they received more services in the other four main areas of production medicine. Grouping of producers, based on whether or not herd records were analyzed by their veterinarian, showed a clustering of adopters of more comprehensive herd health services as clients of two practices. This suggests that comprehensive herd health services are not readily available from all veterinary clinics in the Saskatoon milkshed.

Superovulation in the cow with pregnant mare serum gonadotrophin: effects of dose and antipregnant mare serum gonadotrophin serum.

The effects of pregnant mare serum gonadotrophin (PMSG) dose and PMSG antiserum on superovulation in crossbred beef cows were studied. In experiment I, three groups were treated with 1200, 2400 or 3600 IU of PMSG and 48 h later with prostaglandin (PGF). The mean numbers of corpora lutea (CL), unovulated follicles, and total ova/embryos collected increased as the PMSG dose increased. The percent of fertilized ova and transferable embryos was lowest in the highest dose group (p < 0.05). In experiment II, all cows received 2500 IU of PMSG; groups 1 and 2 were treated with sheep anti-PMSG serum at 48 h or 60 h after PGF; group 3 cows were PMSG-only controls. The number of CL was lowest and the number of unovulated follicles highest in the PMSG-only group (p < 0.05). The number of CL was higher in group 2 (anti-PMSG at 60 h) than in the control group, with the anti-PMSG at 48 h not different from the other groups. Numbers of total ova/embryos, fertilized ova, and transferable embryos were higher (p < 0.05) in both antiserum-treated groups relative to the PMSG-only group. We conclude that superovulation of beef cows with PMSG and treatment with PMSG antiserum will induce a higher superovulatory response and will result in higher CL numbers and fewer unovulated follicles. Further, the variability in the superovulatory response to PMSG treatment was still evident when PMSG antiserum was administered.

Role of luteal regression in embryo death in cattle.

Inseminated crossbred beef cows and heifers were used in 2 experiments to investigate embryo survival after prostaglandin-induced or spontaneous luteal regression. In Experiment 1, luteal regression was induced by an intramuscular (im) injection of cloprostenol (500 mug) on Day 15 (day of ovulation = Day 0). Progestagen was replaced 24 or 36 h later either by one-and-a-half Syncro-Mate-B (SMB) ear implants (9 mg of norgestomet) and norgestomet solution (2.25 mg, im) containing no estradiol in 1 replicate or by 2 SMB implants (12 mg of norgestomet) and progesterone (100 mg, im) in the second replicate. Combined for both replicates, the Day-24 pregnancy rate in an untreated control group (Group 1, 16/19; 84%) was higher (P < 0.01) than in the 24-h group (Group 2, 9/20; 45%), which also was higher (P < 0.02) than in the 36-h group (Group 3, 3/23; 13%). In Experiment 2, 15 or 16 d after breeding, cattle with a corpus luteum at least 16 x 16 mm were given either 2 SMB implants or no treatment. At 24 to 26 d after breeding, pregnancy rates (48/65, 74% versus 49/68, 72%) were not significantly different, and all but 1 of the pregnant progestagen-supplemented cattle had a functional corpus luteum.

Effects of bovine follicular fluid and partially purified bovine inhibin on FSH and LH release by bovine pituitary cells in culture.

We have established a dispersed bovine pituitary cell culture system to study the effects of charcoal-extracted bovine follicular fluid (BFF) or bovine inhibin, partially purified by immunoaffinity chromatography (IPI), on the spontaneous release of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Pituitary cells were plated at 0.25, 0.5 or 1 x 10(6) viable cells/well (c/w) and incubated for 48 h. The medium was replaced and BFF (0, 0.54, 2.7, 13.7, 68.7 or 343.5 micrograms protein) or IPI (0, 0.01, 0.06, 0.29, 1.45 or 7.25 micrograms protein) added to the cultures and the incubation was continued for 48 h. Concentrations of FSH and LH in spent medium were determined by RIA and data analyzed by ANOVA with means compared by Student-Neuman-Keuls (SNK) test. We have shown an increase in spontaneous FSH and LH release attributable to both number of bovine pituitary cells plated and to the length of incubation. The addition of BFF reduced spontaneous FSH release over 48 h incubation. The dose-dependent inhibition curves observed in culture in which different numbers of cells were plated, indicates that inhibition was greater when 1 x 10(6) c/w were plated compared to 0.25 or 0.5 x 10(6) c/w. Bovine follicular fluid at 0.45 micrograms of protein (equivalent to 0.01 microliters of BFF) incubated with 1 x 10(6) c/w, suppressed FSH release by 10.6% compare to control. Maximal suppression of 34.1% was obtained with 50 micrograms (equivalent to 1.56 microliters of BFF). Immunopurified bovine inhibin at 1.45 micrograms tended to suppress FSH release and at 7.25 micrograms significantly reduced FSH release. Neither BFF nor IPI had a measurable effect on LH release. We conclude that BFF and IPI suppress the spontaneous release of FSH from bovine pituitary cells in culture in a dose-dependent manner, without concomitant suppression of LH release.

Superovulation of beef heifers with Folltropin: A new FSH preparation containing reduced LH activity.

The optimum superovulatory dose of Folltropin was determined and compared with a standard 28 mg dose of FSH-P in beef heifers. In Experiment 1, mean numbers of corpora lutea (CL) did not differ among the groups treated with 10, 20, 30 or 40 mg Folltropin or FSH-P, and the mean CL number was reduced (P<0.05) only in the 5 mg Folltropin group. Mean numbers of ova/embryos recovered, fertilized and transferable were greater (P<0.05) for the 10, 20 and 30 mg Folltropin groups than for the 5 mg group. The 40 mg Folltropin group and the FSH-P group were intermediate. The percentage of fertilized and transferable embryos did not differ over the dosages used in this experiment. In Experiment 2, mean numbers of CL were greater for the 9, 18 and 36 mg Folltropin groups than for the 4.5 mg group, with the 9 mg group being lower than the 36 mg group (P<0.05). The 18 mg group was intermediate and did not differ. Mean numbers of ova/embryos recovered and fertilized ova were greater for the 9, 18 and 36 mg groups (P<0.05) than for the 4.5 mg group. The percent of fertilized and mean number and percentage of transferable embryos did not differ among treatments. We conclude that Folltropin may be a satisfactory superovulatory replacement for FSH-P and that a dose of 18 to 20 mg Folltropin may be within the optimum superovulatory dosage range for beef heifers. Dosages of Folltropin of more than twice the optimum did not result in deterioration of ova/embryo quality.

Endocrine and superovulatory responses in heifers pretreated with FSH or bovine follicular fluid.

This study examined the effects of altered serum FSH concentration on subsequent ovarian response to superovulation. Synchronized heifers were assigned randomly on Day 1 of the cycle (estrus = Day 0) to three pretreatment groups that consisted of 6-d of saline (7ml, s.c., b.i.d.; Group I), FSH-P (0.5 mg, i.m., b.i.d.; Group II) or charcoal-extracted bovine follicular fluid (BFF; 7 ml, s.c., b.i.d.; Group III) injections. Superovulation was initiated on Day 7 and consisted of FSH-P in decreasing dosages over 4 d (4,3,2,1 mg; i.m., b.i.d.), with cloprostenol (500 mug) on the morning of the third day. A second replicate with 14 heifers was conducted using the same protocol but twice the pretreatment dosage of FSH-P (1 mg) and BFF (14 ml). Endogenous plasma FSH decreased during BFF and FSH-P pretreatments compared to controls (P < 0.02). Endogenous FSH concentrations in both primed groups (II and III) were similar to control values (Group I) 12 h after the start of superovulation. Basal LH concentrations were not different between pretreatment groups. The interval from cloprostenol treatment to the preovulatory LH surge in Group III was 21.3 and 23.9 h longer (P < 0.0001) than it was in Groups I and II. The postovulation progesterone rise was delayed in Group III. The number of corpora lutea (CL) was lowest in the BFF-primed group (4.2 +/- 0.8) compared with the FSH-primed (7.4 +/- 1.3) and the control (12.0 +/- 1.8; P < 0.003) groups. In the FSH-primed group (0.68 +/- 0.06 cm(3)), CL volumes were larger than in the control group (0.45 +/- 0.03 cm(3)), whereas in the BFF-primed group (0.27 +/- 0.02 cm(3)) CL volumes were smaller compared with the control group (P < 0.0001). Mean FSH concentrations for 48 h preceding superovulation and the number of CL per cow were positively correlated (r = 0.55; P < 0.004; n = 26). We concluded that both FSH-P and BFF pretreatments decreased the superovulatory response of heifers to FSH-P. The mechanism for this would appear to be associated with reduced endogenous FSH prior to the start of superovulation.

Effects of bovine follicular fluid inhibin on serum gonadotrophin concentrations in ewes during oestrus.

Experiments were conducted with ewes to investigate the effects of an enriched bovine follicular fluid inhibin preparation (INH) on gonadotrophin secretion after the onset of oestrus. Administration of INH (10 mg) 1 h after the onset of oestrus did not significantly alter the preovulatory FSH and LH surges or the second FSH peak. To determine the effects of INH on the second FSH surge, ewes were treated with saline (N = 7) or INH (N = 10) at 4 h (10 mg) and 24 h (5 mg) after the peak of the preovulatory LH surge. The second FSH surge was delayed about 24 h (P less than 0.05) in ewes treated with INH; however, the delay did not alter the interval to the next oestrus. In a third experiment, 16 ewes were assigned to 4 groups in a 2 x 2 factorial with the main effects being ovariectomy at 4 h and INH treatment (10 mg) at 4, 20 and 36 h after the peak of the LH surge. Controls received sham ovariectomy and saline injection as appropriate. Ovariectomy resulted in a rapid increase in serum FSH but not LH and this was delayed (P less than 0.05) by INH treatment. These results indicate that inhibin has a selective inhibitory action on FSH secretion in ewes and suggests that the second FSH surge results from increased basal FSH secretion due to decreased endogenous inhibin levels.

Testicular development, daily sperm production and epididymal sperm reserves in 15-mo-old Angus and Hereford bulls: effects of bull strain plus dietary energy.

Bull calves (n = 143) were obtained from two strains of Angus and two strains of Hereford cattle for which replacements were selected on the basis of superior feedlot growth performance on either high- or medium-energy diets. From weaning to slaughter at 15 mo of age, bulls were fed either the high-energy (80% grain + 20% forage) or medium-energy diet (100% forage) corresponding to their strain. Bulls in high-energy diet groups had a greater (P less than .05) scrotal circumference at 12 mo, but not 15 mo of age, than bulls in medium-energy diet groups. Compared with Hereford bulls, Angus had greater (P less than .01) scrotal circumference (36.1 vs 33.9 cm) and greater (P less than .05) paired testes weight (570 vs 464 g) at 15 mo of age. Daily sperm production per gram testicular parenchyma (DSP/g) was affected by strain-diet (P less than .01) but not by breed. Bulls in medium-energy diet groups had 12% greater DSP/g than did high-energy diet bulls (17.4 X 10(6) vs 15.5 X 10(6)). Daily sperm production (DSP) was 9% and 30% greater (P less than .01) for medium-energy diet bulls in 1980 (8.2 X 10(9) vs 7.5 X 10(9)) and 1981 (8.0 X 10(9) vs 6.2 X 10(9)), respectively, compared with high-energy diet bulls. The effect (P less than .01) of breed on DSP was attributed to breed differences in paired testes weight.(ABSTRACT TRUNCATED AT 250 WORDS)

Failure of human chorionic gonadotropin injections to sustain gonadotropin-releasing hormone-induced corpora lutea in postpartum beef cows.

Anestrous postpartum (PP) Hereford cows (n =20) were used to determine the effects of repeated injections of human chorionic gonadotropin (hCG) on the progesterone (P4) secretion and functional lifespan of gonadotropin-releasing hormone (GnRH)-induced corpora lutea (CL). Suckling was reduced to once a day from Day 21 to Day 25 PP, and all cows received injections of 200 micrograms GnRH at 1500 h on Day 24 PP to induce ovulation. Treated cows (HCG, n = 10) received 200 IU hCG b.i.d. from 1900 h on Day 27 PP to 1900 h on Day 33 PP; control cows (CTRL, n=10) were not injected. Blood was collected on Days 21, 23, 25, and 27 to 33, 35, 37, and 39 PP. Serum P4 concentration was measured by radioimmunoassay and used to classify luteal lifespan and the associated estrous cycle as short (SHORT) or normal (NORM) in duration. Treatment with hCG resulted in more (p less than 0.01) cows with SHORT cycles (7 of 9 vs. 4 of 9). Serum P4 concentrations were similar (p greater than 0.20) between groups from 4 days before until 6 days after GnRH injection. Cows with NORM cycles (n = 7) had greater serum P4 concentrations (p less than 0.05) on Days 7 to 11 after GnRH than cows with SHORT cycles (n = 11). By Day 39 PP, all cows with SHORT cycles appeared to have undergone a second ovulation. Charcoal-stripped serum pools from before (PRE) and during hCG injection (INJ) were assayed for total luteinizing hormone-like bioactivity (LH-BA) using a dispersed mouse-Leydig cell bioassay.(ABSTRACT TRUNCATED AT 250 WORDS)

The postpartum induced corpus luteum: functional differences from that of cycling cows and the effects of progesterone pretreatment.

Anestrous postpartum (PP) Hereford cows (n = 41) were used to compare corpora lutea (CL) from gonadotropin-releasing hormone (GnRH)-induced ovulation with CL from cycling cows. Postpartum cows were injected i.m. daily with 100 mg progesterone (P4) or oil on Days 25 through 28 PP and then given 200 micrograms GnRH i.m. on Day 30 PP. Corpora lutea were removed from one-half of the PP cows in the oil- and P4-treated groups 6.5 days after GnRH injection, and from the cycling cows 7 days after estrus. Intact PP cows were used to evaluate cycle length. Blood was collected daily from all PP cows from Day 25 PP through luteectomy and on Days 9, 11, and 13 post-GnRH from the oil- and P4-intact cows to determine short (SHORT) versus normal (NORM) luteal phases. Cycling cows were bled daily from estrus until CL removal NORM PP cows had higher (P less than 0.001) P4 levels than did SHORT PP cows from Day 7 through Day 13 post-GnRH, and more (P less than 0.05) P4-intact cows were NORM compared with oil-intact cows (45.5% vs. 14.3%, respectively). Corpora lutea from cycling cows were heavier (P less than 0.05) and had a higher luteinizing hormone (LH) receptor concentration (P less than 0.05), but CL P4 concentration did not differ from PP cows. Corpora lutea weight, LH receptor and P4 concentration, and in vitro P4 production were similar in the oil-and P4-treated PP cows. NORM cows had heavier CL (P less than 0.05) than SHORT cows, although P4 content and LH receptor concentration did not differ.(ABSTRACT TRUNCATED AT 250 WORDS)

The hypothalamo-pituitary gonadotrophic axis of suckled and nonsuckled dairy cows postpartum.

The objective was to determine whether the suckling-induced delay in return to estrus postpartum could be explained by changes in hypothalamic LHRH content or ability of the pituitary to release LH and FSH in response to LHRH or 59 mM K+ in vitro. In addition, serum concentrations of several other hormones were measured. Nine Holstein cows were suckled ad libitum by two calves and milked by machine twice daily and eight were milked by machine only from calving until slaughter on day 14 postpartum. On day 13 postpartum, blood was collected at 15-min intervals from 0815 to 1200 hr and from 2015 to 2400 hours. Suckled cows had lower (P less than .05) mean serum LH concentrations on day 13 postpartum than did nonsuckled controls. This decrease resulted from a 60% reduction in frequency and a 40% reduction in amplitude of episodic LH peaks. Suckling did not affect body weight change postpartum or serum concentrations of progesterone, estradiol-17 beta, total glucocorticoids, prolactin or FSH during the first 14 days postpartum. The suckling-induced decrease in serum LH was not reflected by a reduction in hypothalamic LHRH or pituitary LH on day 14 postpartum. However, pituitary explants from suckled cows on day 14 postpartum secreted 50% less (P less than .01) LH in response to LHRH (25 ng/ml for 30 min) or K+ (59 mM for 30 min) in vitro than did those from nonsuckled cows. Secretion of FSH was increased 20-fold by LHRH and K+ in vitro, but differences due to suckling treatment were not significant. Decreased frequency and amplitude of episodic LH secretion in vivo and reduced capacity of pituitaries to respond to LHRH may be the cause of suckling-induced inhibition of postpartum ovulation in cattle.

Suckling and four-times daily milking: influence on ovulation, estrus and serum luteinizing hormone, glucocorticoids and prolactin in postpartum holsteins.

Holstein cows were assigned at calving to be (1) milked at 12-hr (n = 5) or (2) 6-hr (n =5) intervals or (3) suckled ad libitum by one calf and milked at 12-hr intervals (n = 6). Suckling resulted in an increase in the postpartum (PP) interval to first ovulation (39.7 vs 21.2 days), but increased milking frequency did not. Delayed resumption of episodic luteinizing hormone (LH) secretion was associated with the increased PP interval to ovulation in suckled cows. Both frequency and amplitude of episodic LH peaks were reduced (P less than .05) on days 7 and 14 PP in suckled cows compared to nonsuckled cows (1.2 vs 2.5 peaks/4.5 hr and 1.8 vs 2.8 ng/ml, respectively). Suckling did not significantly modify basal of milking-induced concentrations of prolactin or total glucocorticoids. Nor did serum concentrations of progesterone or estradiol-17 beta differ between suckled and nonsuckled cows, or between day 7 and day 14 PP. Milking-induced secretion of both prolactin and total glucocorticoids were greater (P less than .05) on day 14 PP than on day 7 PP in both suckled and nonsuckled cows. These results suggest that suckling may delay the first PP ovulation by suppressing episodic LH secretion. Alternations in prolactin, total glucocorticoids, progesterone or estradiol-17 beta do not apper to mediate directly the effects of suckling on PP episodic LH secretion and (or) ovulation.