Evaluation of the effects of sire and dam calving group on age at first calving in Brahman heifers.

This study evaluated the possible effects of sire and dam calving groups on age at first calving in Brahman heifers. A total of 570 heifers born between the years 2004 and 2017 were exposed as yearlings to fertile bulls through time of pregnancy determination. A calving group was determined by calculating the mean (993 d) and standard deviation (187 d) of heifer age at first calving. Heifers considered to calve early (≤899 d; calving group = 1) or late (≥1087 d; calving group = 3) were at least half a standard deviation (94 d) away from the mean. All other heifers were considered to have an intermediate age at first calving (900-1086 d; calving group = 2). Of the 570 heifers, only heifers from a dam (n = 182) with a known age at first calving and from a sire (n = 35) with 5 or more daughters were kept to determine the effect of dam calving group and the effect of sire calving group on age at first calving and calving group in daughters, resulting in a total of 284 heifers available for analysis. Variables included were dam and sire calving groups of the heifer, heifer age at first calving, heifer calving group, heifer season of birth, and heifer year of birth. Data were analyzed using the GLM procedures of SAS and proportions were tested using Chi-square. Sire calving group did affect (P < 0.01) age at first calving and calving group in heifers, but dam calving group did not affect (P > 0.10) daughter age at first calving or calving group. Analysis of dam calving group and sire calving group effects identified a year of birth effect (P < 0.01) on daughter age at first calving and calving group, whereas there was no significant season of birth effect. The proportion of daughters calving early for sire calving groups differed significantly from the expected proportion (P < 0.01), whereas the proportion of daughters calving early for dam calving groups did not differ. An effort to produce a greater proportion of Brahman heifers capable of calving early will not be effective from the dam side but may be effective from the sire side.

Endocannabinoid 1 and 2 (CB(1); CB(2)) receptor agonists affect negatively cow luteal function in vitro.

Thirty to 40% of pregnancies are lost during the first third of pregnancy, which has been hypothesized to be due to inadequate progesterone secretion by the corpus luteum. Loss of luteal progesterone secretion during the estrous cycle is via uterine secretion of prostaglandin F(2)alpha (PGF(2)alpha). Cow luteal tissue secretion of prostaglandins (PG) E (PGE(1)+PGE(2)) and PGF(2)alpha are derived from precursors in membrane phospholipids. Cow luteal tissue secretion of PGE and PGF(2)alpha increased linearly with time in culture with the PGE: ratio being 1:1. PGE(1) or PGE(2) are luteotropic in cows and ewes and antiluteolytic in vitro and in vivo in ewes. Endocannabinoids are also derived from phospholipids and are associated with infertility, presumably by reducing implantation; however, effects of endocannabinoids on luteal function have not been addressed. The objective of this experiment was to determine the effects of endocannabinoid type 1 and 2 receptor agonists and receptor antagonists or a fatty acid amide hydrolase (FAAH; catabolizes endocannabinoids) inhibitor, PGE(1), or PGF(2)alpha on bovine luteal secretion of progesterone, PGE, and PGF(2)alphain vitro. PGE and PGF(2)alpha was increased (P< or =0.05) with time in culture, while progesterone did not change (P> or =0.05) with time in vehicle-treated luteal slices in vitro. Progesterone was increased (P< or =0.05) by PGE(1) and decreased (P< or =0.05) by PGF(2)alpha, CB(1) or CB(2) receptor agonists, or a FAAH inhibitor. Both PGE and PGF(2)alpha were decreased (P< or =0.05) by CB(1) or CB(2) receptor agonists or a FAAH inhibitor when compared to vehicle controls. It is concluded that endocannabinoid receptor agonists negatively affect cow luteal function in vitro and that the corpus luteum may also be a site for endocannabinoid decreased fertility as well as a reduction in implantation.

Relationships between numbers of antral follicles and postpartum interval in Brahman females.

To determine relationships of ovarian antral follicle count (AFC) on d 28 postpartum with postpartum interval (PPI) and AFC on d 8 post-estrus, three-dimensional ultrasonography was performed on multiparous (n = 47) and primiparous (n = 20) Brahman cows on d 28 postpartum and on d 8 post-estrus. Follicles were evaluated to determine numbers of total, small (≤17 mm3), medium (17 mm3-67 mm3), and large follicles (≥67 mm3) on each ovary. Estrus was detected using vasectomized, penile-deviated bulls fitted with chin-ball markers during the AI period and with tail paint during the natural service period. Cows with AFC ≥ ½ SD above the mean were classified as high, those within ½ SD of the mean were classified as intermediate, and those ≥ ½ SD below the mean were classified as low. Classes of AFC for total follicles included low (L ≤ 73), intermediate (74 ≤ I ≤ 112), and high (H ≥ 113). Classes of AFC for small follicles included low (L ≤ 55), intermediate (56 ≤ I ≤ 87), and high (H ≥ 88). Classes of AFC for medium follicles included low (L ≤ 13), intermediate (14 ≤ I ≤ 23), and high (H ≥ 24). Classes of AFC for large follicles included low (L ≤ 2), intermediate (3 ≤ I ≤ 4), and high (H ≥ 5). Cows with PPI ≥ ½ SD above the mean were classified as long, those within ½ SD of the mean were classified as intermediate, and those ≥ ½ SD below the mean were classified as short. Classes of PPI included short (S ≤ 47 d), intermediate (48 d ≤ I ≤ 75 d), and long (L ≥ 76 d). The PROC MIXED procedure of SAS was used to analyze potential differences among classes of AFC and PPI. The PROC REG procedure of SAS was used to analyze relationships between AFC and PPI and between AFC on d 28 postpartum and on d 8 post-estrus. The PPI differed among classes of total (P < 0.01; L = 76.2 ±â€¯6.0, I = 61.7 ±â€¯4.5, H = 43.8 ±â€¯6.5), small (P < 0.01; L = 74.3 ±â€¯5.6, I = 64.6 ±â€¯4.8, H = 42.9 ±â€¯5.9), and medium AFC (P = 0.04; L = 72.3 ±â€¯5.9, I = 59.3 ±â€¯5.8, H = 52.1 ±â€¯5.8). The PPI did not differ among classes of large AFC (P = 0.81). As total AFC on d 28 postpartum increased, PPI decreased (P < 0.01, Adj.R2 = 0.13). Total AFC on d 8 post-estrus was positively associated with total AFC on d 28 postpartum (P < 0.01, Adj.R2 = 0.66). These results suggest that: a) populations of total, small, and medium follicles in the postpartum period predict PPI in Brahman females and b) total AFC in Brahman females are consistent.

Evaluation of the influence of prenatal transportation stress on GnRH-stimulated luteinizing hormone and testosterone secretion in sexually mature Brahman bulls.

This study examined the relationship of prenatal transportation stress (PNS) with exogenous GnRH-induced LH and testosterone secretion in sexually mature Brahman bulls. Brahman cows (n = 96; 48 were stressed by transportation at 5 stages of gestation and 48 were controls) produced a calf crop of 85 calves. All bulls (n = 46) from this calf crop were electroejaculated every 2 wk beginning at a scrotal circumference of 24 cm until sexual maturity (SM; i.e., 500 million sperm/ejaculate). The initial 11 control and 12 PNS bulls to reach SM were selected for the experiment. Within 7-21 d after reaching SM, bulls were fitted with jugular cannulas, from which blood samples were collected at 15-min intervals for 6 h prior to exogenous GnRH administration (10 ng/kg BW; i.v.) and for 6 h after GnRH. Serum concentrations of LH, testosterone, and cortisol were determined by RIA. Age and body weight did not differ ( > 0.1) between PNS and control bulls at the time of the experiment. All bulls responded similarly to exogenous GnRH, indicating no influence of PNS on LH or testosterone response to GnRH. More ( < 0.01) PNS (9 of 11) than control (3 of 12) bulls exhibited an endogenous pre-GnRH LH pulse, and more ( = 0.02) PNS (9 of 11) than control bulls (4 of 12) exhibited a pre-GnRH testosterone response to LH. The average concentration of testosterone during the 60 min (time -60, -45, -30, -15, and 0 min relative to GnRH) immediately preceding GnRH, tended to be greater ( = 0.07) in PNS (1.46 ± 0.30 ng/mL) than control (0.68 ± 0.28 ng/mL) bulls. During that time span serum cortisol was lower ( < 0.01) in PNS (4.00 ± 0.91 ng/mL) than control (7.8 ± 0.87 ng/mL) bulls. A treatment by time interaction ( = 0.03) affected testosterone concentrations from time -240 to 360 min relative to GnRH. Results from this study indicate that PNS did not affect pituitary responsiveness to GnRH or testicular responsiveness to GnRH-induced LH secretion.

Prenatal transportation stress alters temperament and serum cortisol concentrations in suckling Brahman calves.

This experiment examined the relationship between prenatal stress and subsequent calf temperament through weaning. The prenatal stressor used was repeated transportation of pregnant Brahman cows for 2 h at 60 ± 5, 80 ± 5, 100 ± 5, 120 ± 5, and 140 ± 5 d of gestation. Prenatally stressed calves ( = 41) were compared with controls ( = 44; dams did not undergo transportation during pregnancy) from 2 wk of age until weaning (average age at weaning = 174.8 ± 1.3 d). Temperament was defined by pen score (PS; 1 = calm and 5 = excitable), exit velocity (EV; m/sec), and temperament score (TS; (PS + EV)/2) and was recorded for each calf on d -168, -140, -112, -84, -56, -28, and 0 relative to weaning (d 0 = weaning). Cortisol concentrations were determined in serum samples obtained on d -168, -140, -28, and 0 relative to weaning. Birth weight and weaning weight were not different between treatment groups ( > 0.1). Pen score was greater ( = 0.03) in prenatally stressed calves (2.84 ± 0.21) relative to controls (2.31 ± 0.21). Exit velocity was greater ( < 0.01) in prenatally stressed calves (2.1 ± 0.14 m/sec) than in controls (1.61 ± 0.14 m/sec). Exit velocity was affected by a treatment × calf sex interaction ( = 0.04) and was greater in prenatally stressed females. Exit velocity was also affected by day ( < 0.0001). Temperament score was greater ( = 0.01) in prenatally stressed calves (2.45 ± 0.16) than in controls (1.95 ± 0.16). Temperament score was affected by day ( < 0.01). Basal cortisol concentrations were greater ( = 0.04) in prenatally stressed calves (15.87 ± 1.04 ng/mL) than in controls (13.42 ± 1.03 ng/mL). Basal cortisol concentrations were greater ( < 0.01) in females (16.61 ± 1.06 ng/mL) than in males (12.68 ± 1.02 ng/mL). Cortisol concentrations were positively correlated ( < 0.01) with PS ( = 0.55, < 0.01), EV ( = 0.4, < 0.01), and TS ( = 0.55, < 0.01). Overall, suckling Brahman calves that were prenatally stressed were more temperamental and had greater circulating serum concentrations of cortisol than control calves.

Plasma progesterone response following ACTH administration during mid-gestation in the pregnant Brahman heifer.

Previous reports of adrenal progesterone (P4) contributions during late gestation in cattle, and ACTH-induced P4 responses in the non-pregnant heifer, prompted a retrospective investigation to evaluate the plasma P4 response and the relative ratio of plasma cortisol (CT) to P4 following ACTH administration during mid-gestation in pregnant Brahman heifers. Twenty-three pregnant (139.0 +/- 5.0 days of gestation) Brahman heifers received one of the following treatments: 0 (saline; n = 5), 0.125 (n = 4), 0.25 (n = 5), 0.5 (n = 4), or 1.0 (n = 5)IU of ACTH per kg BW. Blood samples were collected at -15 and -0.5 (time 0), 15, 30, 45, 60, 75, 105, 135, 165, 195, and 255-min post-ACTH challenge. Plasma P4 and CT were quantified by RIA. Pre-ACTH P4 did not differ (P > 0.10) among ACTH treatment groups (pooled, 12.1 +/- 0.6 ng/mL). Among peak P4 values at 15-min post-ACTH infusion, control P4 (9.6 +/- 1.2 ng/mL) tended to be lower (P < 0.07) than 0.5 IU ACTH-treated heifers (13.3 +/- 1.1 ng/mL); and were lower (P < 0.02) than 0.25 and 1.0 IU ACTH-treated heifers (14.7 +/- 1.1 and 22.2 +/- 3.7 ng/mL, respectively). During the primary P4 response period (0 to 75-min post-ACTH), the area under the curve (AUC) was greater (P < 0.05) for 1.0 IU ACTH-treated heifers than all other groups. The CT:P4 ratios were lower (time x treatment, P < 0.01) for control heifers than all ACTH-treated heifers. Among ACTH-treated heifers, CT:P4 ratio response and CT:P4 ratio AUC were similar (P > 0.10) following ACTH challenge. In conclusion, acute increases in ACTH elevated plasma P4, likely of adrenal origin, in mid-gestation pregnant heifers, while the CT:P4 ratio (relative output) remained constant irrespective of ACTH dose (0.125-1.0 IU). Whether ACTH-induced increases in P4 in pregnant animals are of physiological significance (e.g., an accessory role in the maintenance of pregnancy during periods of acute stress) remains to be determined.

Physiological and metabolic responses of gestating Brahman cows to repeated transportation.

This study characterized physiological responses to repeated transportation (TRANS) of gestating cows of differing temperaments. Cows were classified as Calm (C; = 10), Intermediate (I; = 28), or Temperamental (T; = 10). Based on artificial insemination date and pregnancy confirmation, cows were TRANS for 2 h on d 60 (TRANS1), 80 (TRANS2), 100 (TRANS3), 120 (TRANS4), and 140 (TRANS5) ± 5 d of gestation. Indwelling vaginal temperature (VT) monitoring devices were inserted 24 h before each TRANS with VT recorded from 2 h before TRANS and averaged into 5-min intervals through 30 min after TRANS. Serum samples were collected before loading and on unloading from the trailer to determine concentrations of cortisol, glucose, and nonesterified fatty acids (NEFA). Data were analyzed by repeated measures analysis in SAS. Serum cortisol concentrations were affected by temperament ( < 0.001), with T cows having the greater concentrations of cortisol before each TRANS event. All cows (100%) regardless of temperament exhibited elevations in cortisol following each TRANS event. Peak VT was greater ( < 0.001) at TRANS1 relative to all other TRANS events regardless of cow temperament. During TRANS, the T cows tended ( < 0.09) to have greater peak VT (39.86 ± 0.15°C) compared to C (39.41 ± 0.16°C) and I cows (39.55 ± 0.08°C). Area under the VT curve decreased ( = 0.002) from TRANS1 through TRANS5. Pre-TRANS serum glucose concentration at TRANS1 was greater ( < 0.03) for T (68.13 ± 4.31mg/dL) compared to I (53.42 ± 2.78 mg/dL) and C cows (52.76 ± 4.60 mg/dL). The C and I cows had greater changes in NEFA concentration between pre- and post-transport, and T cows showed the least change ( < 0.001). Cow VT and serum glucose concentration decreased in all temperaments ( < 0.01) with repeated TRANS; however, serum NEFA concentration post-TRANS did not vary ( > 0.10) with repeated TRANS events. Serum glucose concentrations were affected ( < 0.02) by a TRANS event by temperament interaction with T cows taking more TRANS events to decrease their change in glucose concentration compared to C and I cows. These results demonstrate that temperament influences physiological responses to stress in gestating Brahman cows. Although repeated transport in our study is confounded with day of gestation, seasonal changes, and learning from repeated handling and transport, repeated transport is a useful model of repeated stress in studying the effects of temperament.

The effect of aspirin administration and parity on plasma salicylate concentrations and postpartum reproductive parameters in Brahman cows.

Forty pluriparous (M) and 20 primiparous (P) suckled Brahman cows were used to evaluate the effect of aspirin and parity on plasma 13,14-dihydro-15-keto-prostaglandin F2alpha (PGFM) and progesterone (P4) concentrations and some reproductive parameters. On Day 7 after calving (PP), the cows were allocated within parity into 2 groups: the aspirin group received concentrate containing aspirin at a rate of 100 mg/kg of body weight every 12 h until Day 13 PP; and the control received concentrate every 12 h for the same interval. Blood samples were collected after first and last aspirin feeding and daily from Day 1 PP to Day 6 PP and from Day 14 PP to Day 21 PP, twice daily from Day 7 PP to Day 13 PP, and weekly until first non-return to estrus. Plasma salicylate concentrations in the aspirin group cows were affected by parity (P < 0.01) and time after feeding (P < 0.0001). P cows showed higher plasma salicylate concentrations with a later peak and slower decrease than M cows. Aspirin-treated P cows had longer PP intervals than either control P, control M, or aspirin-treated M cows. Cows receiving aspirin had a lower pregnancy rate, an increased incidence of abnormal estrous cycles, and a decline in the presence of corpora lutea after estrus. Cows that formed a corpora lutea and had received aspirin had higher P4 release between Day 6 and 14 after estrus. Aspirin-treated cows that did not form corpora lutea had lower P4 release between Days 9 and 14 after estrus. A treatment by parity interaction affected mean PGFM proportions (P < 0.01) during the treatment period. Aspirin-fed P cows increased PGFM release as measured by mean proportion of Day 6 PP values. Aspirin-fed M cows showed a decrease in mean PGFM proportions. Aspirin feeding during the early PP showed different effects on some reproductive parameters in P and M Brahman cows, indicating differences in PP physiology between parities.

Effects of estrous synchronization on response to nitric oxide donors, nitric oxide synthase inhibitors, and endothelin-1 in vitro.

Two experiments were conducted to determine the effects of nitric oxide (NO) donors, endothelin-(ET-1), and NO synthase (NOS) inhibitors on bovine luteal function in vitro. In experiment 1, estrus in Brahman cows was synchronized with Synchro-Mate-B (SMB) and day-13-14 corpora luteal slices were weighed, diced and incubated in vitro. Treatments (100 ng/ml) were: vehicle, N[see symbol in text]-nitro-L-arginine-L-methyl ester (L-NAME), N(G)-monomethyl-L-arginine acetate (L-NMMA), diethylenetriamine (DETA), DETA-NONOate, sodium nitroprusside (SNP), or ET-1. In experiment 2, estrus was synchronized with Lutalyse, a Controlled Intravaginal Progesterone Releasing Device (CIDR), or cows were not synchronized. Corpora lutea were collected, weighed, and luteal slices were weighed, diced and incubated in vitro with treatments. Treatments (100ng/ml) were: vehicle, L- NAME, L-NMMA, DETA, DETA-NONOate, sodium nitroprusside, S-nitroso-N-acetylpenicillamine (SNAP) or endothelin-1. Tissues were incubated in M- 199 for 1 h without treatments and for 4 and 8 h in both experiments with treatments in both experiments. Media were analyzed for progesterone, prostaglandins E2 and F2alpha (PGE2, PGF2alpha) by radioimmunoassay (RIA). Hormone data in experiments 1 and 2 were analyzed by 2 x 7 and 3 x 2 x 8 factorial design for analysis of variance (ANOVA), respectively. Luteal weights in experiment 2 were analyzed by a one-way ANOVA. Concentrations of progesterone in media were similar (P > or = 0.05) among treatments within experiments. Concentrations of PGE2 in media in experiment 1 were undetectable in 90 and 57% of the samples at 4 and 8 h, respectively. PGF2alpha increased (P < or = 0.05) with time, but did not differ (P > or = 0.05) among treatments. Secretion of PGF2alpha was not affected by treatments (P > or = 0.05). In experiment 2, luteal weights of the induced estrous cycle were decreased (P < or = 0.05) by Lutalyse. Concentrations of PGE2 and PGF2alpha increased (P < or = 0.05) with time in control of all three synchronization regimens. DETA-NONOate, SNAP, sodium nitroprusside (NO donors) and ET-1 increased (P < or = 0.05) PGE2 except in the CIDR synchronized group (P > or = 0.05). No treatment increased (P > or = 0.05) PGF2alpha in any synchronization regimen. It is concluded that either SMB containing norgestomet or a CIDR containing progesterone alters luteal secretion of PGE2, Lutalyse lowers luteal weights in the induced estrous cycle, and NO or ET-1 given alone are not luteolytic agents. It is suggested that NO and ET-1 could have indirect antiluteolytic/luteotropic effects via increasing PGE2 secretion by luteal tissue rather than being luteolytic.

Plasma 13,14-dihydro-15-keto-prostaglandin F2 alpha concentrations in prepubertal dairy heifers challenged with oxytocin.

Twenty-nine prepubertal Holstein heifers were assigned by age to one of three age groups to determine if the prepubertal bovine uterus could respond to an oxytocin stimulus. Group 1 heifers were 6 to 7 months of age (AGE1; n = 11), group 2 heifers were 8 to 9 months of age (AGE2; n = 11) and group 3 heifers were 10 to 11 months of age (AGE3; n = 7). Blood samples were collected via an indwelling jugular catheter. Four samples were collected at 15-min intervals prior to oxytocin administration to determine basal 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) concentrations. Each heifer received 100 IU of oxytocin i.v., blood sampling continued at 5 min intervals for the next 30 min and for an additional 90 min at 15-min intervals. Heifers were considered responders to oxytocin if mean PGFM concentrations increased at least 1.5 times the SD of their basal PGFM concentration. Age of the heifer (P less than .0001) and responder status (P less than .05) affected plasma PGFM. Plasma PGFM was higher in AGE1 and AGE3 heifers than AGE2 (P less than .0001). The number of responders was greatest at AGE3 (P less than .03) with AGE1 and AGE2 being similar. Mean basal PGFM was lower (P less than .04) at AGE2 than AGE1 with AGE3 being intermediate. In addition, basal PGFM at AGE1 tended to be lower (P less than .08) in the responders than in the non-responders, while AGE2 basal PGFM did not differ between responders and non-responders (P greater than .10).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of prenatal stress on the fetal calf.

Twelve pregnant Brahman cows were randomly assigned to one of two treatment groups: 1) transported in a stock trailer for 24.2 km, unloaded at a second farm and penned for 1 hr, and then returned to the original farm (TRANS, n = 6); or 2) walked through the handling facilities (SHAM, n = 6). Treatments were repeated at 60, 80, 100, 120, and 140 d of gestation. Calves were delivered by cesarean section on d 266 of gestation. The male:female ratio was 4:2 and 5:1 for the TRANS and SHAM treatment groups, respectively. Before calf removal and severance of the umbilical blood flow, a blood sample was collected from the calf to determine plasma concentrations of adrenocorticotropin (ACTH) and cortisol. The calf was then sedated and exsanguinated, after which pituitary and adrenal glands were collected. The adrenals were immediately weighed, and a cross-section from the left adrenal was stored in 4% paraformaldehyde until being embedded in paraffin. Eight sections from each adrenal were sliced (5 microns), fixed, and then stained with Harris' hematoxylin and eosin. Areas of the cortex and medulla were calculated with a computerized digitizing unit and tracing of the viewed section. The TRANS calves had heavier body weights (BW) (28.7 vs. 23.9 +/- 1.8 kg; P < 0.07), pituitary glands (12.63 vs. 8.24 +/- 1.10 g/kg BW; P < 0.008), and heart weights (5.58 vs. 5.17 +/- 0.58 g/kg BW; P < 0.05) than did the SHAM calves. Plasma concentrations of ACTH and cortisol did not differ between SHAM and TRANS calves (57 vs. 82 +/- 14 pg/ml and 7.0 vs. 6.7 +/- 0.9 ng/ml, respectively; P > 0.2). Adrenal gland weight and medulla-to-cortex ratio did not differ between SHAM and TRANS calves (0.61 and 0.73 +/- 0.03 g and 0.97 and 0.99 +/- 0.12 g, respectively; P > 0.2). These results suggest that the altered response to stress in prenatally stressed calves is not associated with morphological changes in the adrenal gland but may be due to effects of prenatal stress on the fetal pituitary.

Effect of estrus synchronization with Syncro-Mate-B((R)) on serum luteinizing hormone, progesterone and conception rate in Brahman heifers.

Twenty-two estrous cyclic, 2-yr-old Brahman heifers were randomly assigned to receive either estrus synchronization with Syncro-Mate-B((R)) (SMB; 11) or no treatment (Control; 11). Blood samples were collected via tail vessel puncture at onset of estrus and daily thereafter until Day 11 after estrus. Blood samples were also collected from five SMB and five Control heifers at 0, 4, 8 and 12 h after the onset of estrus. All samples were processed to yield serum and stored at -20 degrees C until radioimmunoassay. Heifers were inseminated by one technician using semen from a single ejaculate of a Brahman bull 12 h after the onset of estrus. All SMB heifers exhibited estrus within 72 h of implant removal. All heifers had corpora lutea (CL) detected by rectal examination 8 to 12 d following estrus. Serum luteinizing hormone (LH) was not affected by treatment, time (4 - h intervals) or an interaction of treatment by time (P > 0.10). Independent analysis with h indicated that at h 12, SMB (2.2 +/- 0.06 ng/ml) had lower LH than did control heifers (8.9 +/- 2.1 ng/ml). Serum progesterone increased from Day 1 through Day 12 in all heifers, which is indicative of functional CL. Serum progesterone was affected by treatment (P < 0.0001) and time (d intervals; P < 0.10). Progesterone elevation was lower (P < 0.05) and area under the progesterone curve was lower (P < 0.03) in SMB (5.6 +/- 0.5 ng/ml, 32.0 +/- 4.5 units, respectively) when compared with control heifers (7.0 +/- 4 ng/ml, 43.7 +/- 2.4 units, respectively). Conception rate was lower (P < 0.01) in SMB heifers (2 of 11) than in control heifers (8 of 11). The lowered conception rate in SMB treated Brahman heifers may be due to altered timing of LH release following estrus, resulting in an altered time of ovulation.

Effects of naloxone and animal temperament on serum luteinizing hormone and cortisol concentrations in seasonally anestrous Brahman heifers.

The effect of endogenous opioid peptides (EOP) and individual animal temperament on serum luteinizing hormone (LH) were investigated in seasonally anestrous Brahman heifers (n = 24). Animals that had shown behavioral estrus in previous months but that had not returned to estrus for at least 30 d were selected. The heifers were ranked by temperament (tame = 1, normal = 2, wild = 3) and randomly allotted into three groups. Blood was collected from one heifer of each group per day. Blood samples were taken via jugular cannula every 15 min for 6 h and every 30 min for another 4 h. After the first hour of sampling, the heifers received intravenous saline (SAL, n = 8); naloxone (LN, 0.5 mg/kg i.v., n = 8); or naloxone (HN, 1.0 mg/kg i.v., n = 8). Three hours after naloxone treatment, each heifer was given gonadotropin releasing hormone (GnRH, 100 microg i.m.). All samples were processed to yield serum and were assayed for LH by radioimmunoassay (RIA). Hourly samples were assayed for cortisol by RIA. The area under the LH curve 60 min postnaloxone treatment was higher in LN and HN than in SAL (57.0 and 40.8 vs 6.1 units; P<0.01); and the area under the 180 min postnaloxone curve remained higher in LN than in SAL (106.2 vs 35.1 units; P<0.05). Cortisol concentrations 60 min postnaloxone administration were above prenaloxone levels(38.2 vs 26.7 ng/ml; P<0.0002). Temperament scores of heifers were positively correlated with cortisol release. The area under the cortisol curve had a negative correlation with mean LH. Serum LH concentrations appear to be suppressed by EOP in seasonally anestrous Brahman heifers, and EOP appear to reduce serum cortisol concentrations. Excitable heifers had higher concentrations of serum cortisol, which negatively affected serum LH concentrations.

Seasonal variations in characteristics of estrous cycles in pubertal Brahman heifers.

A group of pubertal Brahman heifers (n = 16) was monitored from October through March to investigate the seasonal changes in estrous cyclicity. The heifers had a mean age of 16.7 +/- 0.3 mo at the initiation of the experiment. They were kept on pasture with vasectomized marker bulls. Supplemental feed to meet NRC requirements was provided. Estrus occurrence was checked once a day and blood samples were taken weekly by tail venipuncture from heifers that had been in estrus 7 to 14 d earlier. Samples were processed to yield serum and were assayed for serum progesterone by radioimmunoassay. A high proportion of heifers (88%) had abnormalities such as estrus without the formation of a functional corpus luteum (CL) or anestrus, with a distribution of the two abnormalities as follows: October 0 and 0, November 50 and 25, December 0 and 50, January 0 and 50, February 18 and 31 and March 0 and 7%, respectively. Mean serum progesterone concentrations during the luteal phase differed by month: 4.26, 1.50, 3.25, 2.27, 2.65 and 3.70 ng/ml for October, November, December, January, February and March, respectively (P<0.001). Heifers that went into anestrus had lower mean serum progesterone concentrations than heifers that had regular estrous cycles throughout the study period (1.35 vs 2.22 ng/ml; P<0.0005). The months with the shortest daylengths (December and January) had the highest incidence of anestrus. Transitional periods (November and February) seemed to occur before and after the months with the highest occurrence of anestrus. During this transitional period a high incidence of estrus without the formation of a functional CL was detected. Serum progesterone concentrations were lower in all heifers during the months with high occurrence of abnormal estrous cycles.

Effect of alfaprostol on postpartum reproductive efficiency in brahman cows and heifers.

Brahman cows (n=54) and heifers (n=18) were randomly allotted by calving date, sex of calf and age to one of four treatment groups. Group 1 received no treatment (control), Group 2 received 5 mg alfaprostol (AP) i.m. on Day 21 postpartum, Group 3 received 5 mg AP i.m. on Day 32 postpartum and Group 4 received 5 mg AP i.m. on both Days 21 and 32 postpartum. Blood samples were collected via tail vessel puncture at 30 min-intervals for 8 h from half the animals in each group on Days 21 and 32 postpartum, with AP injection administered 2 h after sampling had begun. All cows were bled at weekly intervals. Samples were processed to yield serum and stored at -20 degrees C until assayed for luteinizing hormone (LH) or progesterone (P4). All cattle were maintained with epididymectomized marker bulls and were artificially inseminated (A.I.) at first estrus. Serum P4 was below 1 ng/ml prior to AP treatment in all animals and did not differ (P>0.10) between treatments. Alfaprostol treatment affected mean postpartum interval (from parturition to return to standing estrus and subsequent corpus luteum formation with serum progesterone concentrations>1 ng/ml; P<0.08). The control group (84.8+/-7.9 d) did not differ from Group 2 (86.3+/-11.1 d) or Group 3 (66.7+/-5.5 d) but did differ (P<0.09) from Group 4 (65.1+/-6.4 d). Cattle injected on Day 32 had a shorter (P<0.01) postpartum interval than those not receiving treatment on that day (65.9+/-4.2 vs 85.7+/-6.8 d). Pregnancy rate was affected (P<0.05) by AP treatment. The control group (72.2%) did not differ (P>0.10) from any group but, Group 2 (50.0%) was lower (P<0.04) than Group 3 (83.3%) and (P<0.02) Group 4 (88.9%). Cattle treated on Day 32 (Groups 3 and 4) had a higher (P<0.02) pregnancy rate (86.1%) than those not treated on Day 32 (Groups 1 and 2; 61.1%). Serum LH was affected by day (P<0.0003) and treatment by day (P<0.07) but not by time (P>0.10). Treatment Group 3 (P<0.08) and Group 4 (P<0.0003) mean LH concentrations differed between Days 21 and 32 postpartum. Cattle receiving AP treatment on Day 32 postpartum had a higher (P<0.04) cumulative frequency of return to estrus by 100 days postpartum than nontreated cattle.

Effect of alfaprostol, lasalocid, and once-daily suckling on postpartum interval in Brahman and Brahman crossbred cattle.

Brahman cows (n = 49) and primiparous heifers (n = 11), Brahman x Hereford primiparous F1 heifers (n = 86) and Simmental x Brahman primiparous F1 heifers (n = 13) were randomly allotted by breed, age and date of calving to one of eight treatment groups: 1) control; 2) once-daily suckling; 3) lasalocid (200 mg/hd/d); 4) alfaprostol (5 mg intermuscular injections on Days 21 and 32 post partum); 5) lasalocid + once-daily suckling; 6) alfaprostol + once daily suckling; 7) alfaprostol + lasalocid; 8) alfaprostol + lasalocid + once daily suckling. All animals received 2.3 kg/hd/d of a concentrate (6 corn : 1 cottonseed meal) and lasalocid was mixed and fed in the concentrate. Body weights and condition scores were taken on Day 1 post partum and every 28 d thereafter. All animals were maintained with sterile marker bulls with Brahman and Simmental x Brahman cattle artificially inseminated at first estrus. Blood samples were collected at weekly intervals starting on Day 21 post partum until estrus and at nine to twelve days post estrus when the ovaries were palpated for corpora lutea. After the first postpartum estrus with a corpora lutea, cows were placed with fertile bulls. Mean serum progesterone concentrations were below 0.5 ng/ml prior to treatment. Calf weight gains to 90 d were not affected by age (P > 0.10) but were lower in the once-daily suckling group (P < 0.05). Treatment did not affect cow weight or condition score (P > 0.10). Cows had a shorter postpartum interval (P < 0.0001) than heifers. Once-daily suckling shortened postpartum interval (P < 0.0001) and positively influenced the cumulative frequency of return to estrus by 40 d post partum (P < 0.02). Alfaprostol did not affect postpartum interval (P > 0.10) but did increase the cumulative frequency of return to estrus by 90 d post partum (P < 0.03). Lasalocid did not affect postpartum interval or cumulative frequency of return to estrus (P > 0.10). Both once-daily suckling and alfaprostol were effective in increasing the numbers of animals inseminated by 90 d post partum. The once-daily suckling + alfaprostol treatment resulted in the shortest postpartum interval.

Concentrations of 13, 14-dihydro-15-keto-prostaglandin F(2)alpha, estradiol-17beta and progesterone during the peripubertal period in heifers.

Twenty prepubertal Holstein heifers were utilized to assess plasma 13, 14-dihydro-15-keto-prostaglandin F(2)alpha (PGFM), serum progesterone (P(4)) and estradiol-17beta (E(2)) concentrations as well as the E(2):P(4) ratio during the onset of puberty in cattle. All animals were maintained as a group along with a sterile marker bull to assist in the detection of estrus. Upon detection of the first estrus (Day=O), daily blood samples were collected from a jugular vein until the heifers had completed 3 estrous cycles. The average body weight and age at first estrus were 247.6+/-4.8 kg and 304.0+/-7.5 days, respectively. Frequency of abnormal length estrous cycles was greater (P<0.02) during the first (40%) and second (35%) cycles than during the third estrous cycle (0%). All heifers had normal cycle lengths (18 to 24 days) by the third estrous cycle. Serum P(4) was greater during the third cycle (P<0.05) from Day 10 to Day 4 before the next estrus compared with the same period of the first estrous cycle. Serum E(2) did not peak until the day of estrus in the first cycle, whereas E(2) reached a maximal level 2 days before estrus in the third estrous cycle. Serum E(2) was higher (P<0.0001) 2 days before estrus in the third cycle than in the first estrous cycle. Plasma PGFM reached maximum concentrations 3 days before estrus in the third cycle compared with 1 day before estrus at the end of first estrous cycle. As estrus approached during the third cycle, PGFM rose 1 day before E(2) rose and P(4) declined, while the rise in PGFM and E(2) occurred simultaneously, with P(4) declining at the end of the first estrous cycle. During diestrus, the E(2):P(4) ratio was lower (P<0.07) in the third cycle than in the first, but it was higher (P<0.04) at estrus and 1 day before in the third estrous cycle. These data reveal a high incidence of abnormal length estrous cycles during the first two estrous cycles of the peripubertal period, and demonstrate anomalies in uterine and ovarian endocrine activity during the peripubertal period in cattle.

Testicular and epididymal function during the peripuberal period in Brahman bulls receiving various amounts of protein degradable in the rumen.

Thirty-nine Brahman bulls with an initial age and weight of 301.7 +/- 4.1 d and 202.7 +/- 4.7 kg, respectively, were randomly allocated to 1 of 2 dietary treatment groups within age, weight and sire in order to study the influence of source of protein and stage of peripuberal period on testicular and epididymal function. In the soybean meal treatment the amount of protein undegradable in the rumen averaged 47%, while it was 72% in the fish meal treatment. The supplements were isocaloric and isonitrogenous. Bulls were electroejaculated, and castrations were performed randomly in a predetermined order when the first ejaculate with the first motile sperm cells (Stage 1), 10 to 25 million (Stage 2), and 50 million or more sperm cells (Stage 3 - puberty) was obtained. Testicular and epididymal traits were analyzed for a single testicle and epididymis. Daily sperm production, daily sperm production per gram of testicular parenchyma, testicular weight and testicular parenchyma weight were not affected by treatment. Bulls receiving fish meal had heavier (P < 0.01) epididymis than soybean meal-fed bulls (6.6 +/- 1.0 vs 3.9 +/- 0.6 g) but similar (P > 0.05) epididymal sperm reserves. Daily sperm production (1 testicle) was 115.2 +/- 0.1, 447.4 +/- 0.1, 792.7 +/- 0.1 million sperm cells, and daily sperm production per gram of testicular parenchyma was 1.5 +/- 0.5, 3.2 +/- 0.6 and 6.4 +/- 0.6 million sperm cells for bulls at Stage 1, 2 and 3, respectively. Sire and amount of undegradable intake protein had significant (P < 0.05) affects on the distribution of epididymal sperm reserves, with soybean meal-fed bulls having the higher proportions of epididymal sperm reserves in the cauda epididymis.

Exogenous PGF(2)alpha enhanced GnRH-induced LH release in postpartum cows.

This study evaluated the effect of exogenous PGF(2)alpha on circulating LH concentrations in postpartum multiparous (n = 32) and primiparous (n = 46) Brahman cows. The cows were randomly allotted within parity and calving date to receive 0, 1, 2 or 3 mg im PGF(2)alpha (alfaprostol)/100 kg body weight (BW), with or without GnRH on Day 30 after calving. Blood samples were collected at weekly intervals from calving through treatment. Serum progesterone concentrations were determined using RIA procedures to assure that only anestrous cows were treated. Sterile marker bulls were maintained with cows on Coastal bermudagrass pastures until the first estrus was detected. Multiparous cows had a shorter (P < 0.05) interval from calving to estrus than did primiparous cows. Serum LH was affected by time (P < 0.0001), PGF(2)alpha dose (P < 0.0002), GnRH (P < 0.0001), parity by PGF(2)alpha dose (P < 0.0003), PGF(2)alpha dose by GnRH (P < 0.0009), parity by GnRH (P < 0.0008), and by parity by PGF(2)alpha dose by GnRH (P < 0.0005). Multiparous cows not receiving GnRH had higher mean serum LH (P < 0.02), LH peak pulse height (P < 0.03), and area under the LH release curve (P < 0.03) compared with primiparous cows. The number of LH pulses/6 h was greater (P < 0.06) in multiparous than primiparous cows, and was greater (P < 0.02) in multiparous cows receiving 3 mg/100 kg BW than in cows receiving 2 mg/100 kg BW, but not in the controls or in cows receiving 1 mg/100 kg BW. Exogenous GnRH resulted in increased (P < 0.0001) serum LH concentrations in all cows, and LH was enhanced (P < 0.0009) by simultaneous treatment with PGF(2)alpha. Primiparous cows had a greater response (P < 0.0005) to PGF(2)alpha and GnRH compared with multiparous cows. Pituitary release of LH in response to GnRH was enhanced by simultaneous exposure to PGF(2)alpha in Day 30 postpartum cows.

Artificial insemination following observational versus electronic methods of estrus detection in red deer hinds (Cervus elephus).

The objectives were to determine the efficacy of the HeatWatch (HW) electronic estrus detection system for monitoring behavioral estrus (including duration and intensity) in red deer hinds and to evaluate pregnancy rate to AI after detected estrus. Red deer hinds (Cervus elephus; n = 50) were allocated into two treatment groups: AI following synchronization (CIDR/PMSG) and observed estrus (induced estrus group: IE; n = 25) or AI following the detection of natural estrus (NE; n = 25) without hormonal treatment. Hinds were fitted with two HeatWatch (HW) electronic estrus detection transmitters, one above the tail (bottom) and one between the tuber coxae of the pelvic girdle (top), and visual observations for mounting activity began with the aid of young sterile red deer stags (18 months old) fitted with marking harnesses. Hinds in both groups were inseminated (10-12h after observed estrus) with frozen-thawed red deer semen using a transvaginal/cervical AI approach. Following a 26-day period of AI, hinds were placed with a mature fertile stag for an additional 30-day natural breeding period. Pregnancy diagnosis was performed 57 and 86 days after the start of AI. While the hinds were housed with the young stags, 82% were detected in estrus by visual appraisal of stag crayon marks, but only 32% of these were detected by HW. In contrast, in the hinds housed with the mature stag, 93% detected in estrus by crayon marks were also detected by HW. The top HW transmitter consistently recorded more mounts (P < 0.05) than the bottom transmitter. The pregnancy rate was numerically better in IE versus NE hinds (42% versus 29%, P > 0.10). In summary, there were no differences (P > 0.10) in the intensity (number) or duration of mounts (detected by HW) during estrus in IE versus NE hinds, and HW was most effective in detecting estrus in the presence of a heavier, mature stag versus a younger stag. When used in combination with transvaginal AI, an overall first-service pregnancy rate of 36.6% was achieved with AI of frozen-thawed semen in red deer hinds following detected estrus.