Mark A Crowe

Abstract: The oestrous cycle in cattle lasts for 18-24 days. It consists of a luteal phase (14-18 days) and a follicular phase (4-6 days). During the cycle there are generally two (dairy cows) or three (heifers and beef cows) waves of ovarian follicle growth. Each wave of follicle growth consists of a period of emergence of a cohort of follicles, selection of a dominant follicle and either atresia or ovulation of the dominant follicle. These waves of follicle growth, initially established during the early pre-pubertal period of development occur throughout the entire cycle, with only the dominant follicle (DF) of the final wave coinciding with the follicular phase that undergoes final maturation and ovulation. Ovarian functions (follicle growth, ovulation, luteinisation and luteolysis) are regulated by the endocrine hormones of the hypothalamus (gonadotrophin-releasing hormone), anterior pituitary (follicle-stimulating hormone and luteinising hormone), ovaries (progesterone, oestradiol and inhibins) and the uterus (prostaglandin F2α). In postpartum cows resumption of regular oestrous cycles (in addition to uterine involution) is fundamental for re-establishment of pregnancy.

Abstract: In cattle, elevated concentrations of circulating progesterone (P4) in the immediate postconception period are associated with advanced conceptus development, while low P4 is implicated as a causative factor in low pregnancy rates observed in dairy cows. This study aimed to: 1) describe the transcriptional changes that occur in the bovine endometrium during the estrous cycle, 2) determine how elevated P4 affects these changes, 3) identify if low P4 alters the expression of these genes, and 4) assess the impact that low P4 has on conceptus development. Relatively few differences occurred in endometrial gene expression during the early luteal phase of the estrous cycle (Day 5 vs. 7), but comparison of endometria from more distant stages of the luteal phase (Day 7 vs. 13) revealed large transcriptional changes, which were significantly altered by exogenous supplementation of P4. Induction of low circulating P4 altered the normal temporal changes in gene expression, and these changes were coordinate with a delay in the down-regulation of the PGR from the LE and GE. Altered endometrial gene expression induced by low P4 was associated with a reduced capacity of the uterus to support conceptus development after embryo transfer on Day 7. In conclusion, the present study provides clear evidence that the temporal changes in the transcriptome of the endometrium of cyclic heifers are sensitive to circulating P4 concentrations in the first few days after estrus. Under low P4 conditions, a suboptimal uterine environment with reduced ability to support conceptus elongation is observed.

Abstract: The objective was to investigate if Banding or Burdizzo castration of bulls would alter the gene expression profile of a range of peripheral leukocyte inflammatory cytokines (IL-1, IL-6, IL-8, IL-10, interferon-γ and tumor necrosis factor-α) and to determine if the administration of carprofen (C) before castration would affect the expression of these genes. Thirty Holstein-Friesian bulls (5.5 months; Mean 191±(SEM) 3.7 kg) were blocked by weight and randomly assigned to one of five treatments: (1) untreated control (CON); (2) Banding castration at 0 min (BAND); (3) BAND following an i.v. injection of 1.4 mg/kg BW of carprofen (C) at -20 min (BAND+C); (4) Burdizzo castration at 0 min (BURD); or (5) BURD following 1.4 mg/kg BW of carprofen at -20 min (BURD+C). Blood samples were collected at 1 h before castration and 6, 24 and 48 h post-castration for routine hematology and quantitative real-time PCR analysis of cytokine gene expression analysis. Generally, there were no differences (P>0.05) among treatment groups in hematological variables following castration. Cortisol concentrations were unchanged throughout the experimental period in CON bulls. BURD animals had greater cortisol concentrations than BAND and CON animals at 6 h post treatment. Transitory effects were observed only in the expression of IL-6 and TNF-α. The relative expression of IL-6 was greater in the BURD than in the BAND treatment (P<0.05) at 24 h post-castration and was greater in the BURD+C group than in the BURD group (P<0.05) at 48 h. The relative expression of TNF-α was greater in BAND than in the BURD group (P<0.05) at 48 h. In conclusion, these findings indicate that Banding or Burdizzo castration did not have any major effect on peripheral leukocyte inflammatory cytokine gene expression; Banding castration caused a greater pro-inflammatory cytokine gene expression reaction than Burdizzo castration and carprofen administration can affect IL-6 gene expression levels in BURD castrated animals.

Abstract: Progesterone induced blocking factor (PIBF) and galectins modulate the maternal immune response during pregnancy. We hypothesized that the relative transcript abundance of the above genes would be different during the luteal phase/early pregnancy, and would be affected by progesterone supplementation. To further test this hypothesis protein expression analyses were carried out to evaluate the abundance, and localization of LGALS9 and PIBF. Following oestrus synchronization, heifers were either inseminated (n=140) or not (n=70). Half of the heifers in each status (cyclic or potentially pregnant) were randomly assigned to receive a progesterone releasing intra-vaginal device (PRID) on Day 3 after oestrus, which elevated progesterone concentrations from Day 3.5 to 8 (p<0.05), resulting in four treatment groups: cyclic and pregnant heifers, each with normal and high progesterone. After confirmation of pregnancy status in inseminated animals, uterine tissue was collected on Days 5, 7, 13 or 16 of the luteal phase of the cycle/pregnancy. Gene and protein expression was determined using Q-RT-PCR and IHC, respectively, on 5 heifers per treatment per time point (i.e., 80 in total). Progesterone concentrations did not affect expression of any of the genes (p>0.05). LGALS9 and LGALS3BP were expressed at low levels in both cyclic and pregnant endometria until Day 13. On Day 16, expression increased only in the pregnant heifers (p<0.0001). LGALS1 and LGALS3 decreased on Day 7 (p<0.0001), and remained low until Day 16. Pregnancy had no effect on the expression of LGALS1, LGALS3 and PIBF. Additionally, LGALS9 and PIBF proteins were expressed in distinct uterine cell types. These results indicate that the galectins may be involved in uterine receptivity and/or implantation in heifers.

Abstract: Improving the energy status of dairy cows during the early post-partum (PP) period by adding a safe dietary supplement such as live yeast culture (YS) may have a positive effect on reproductive function. The objective was to examine potential benefits of YS supplementation on PP energy status and fertility indices of dairy cows managed to have low or high body condition score (BCS, 1-5 scale) at calving. Forty (10 primiparous and 30 multiparous) Holstein/Friesian dairy cows were blocked by yield, parity, BCS, and predicted calving date. Within each block, cows were randomly allocated to a 2 x 2 factorial arrangement of treatments which were: BCS at calving (low < or =3.5 or high > or =3.75; n=20) and YS supplementation (2.5g/cow/day for pre-calving and 10g/cow/day for post-calving x 10(8) CFU of Saccharomyces cerevisiae/g) (supplemented or control; n=20). Daily milk yield was recorded and weekly milk composition, BCS and BW were assessed from calving to week 10 PP. Forage (100% grass silage pre-calving; 50% grass silage, 50% maize silage post-calving; ad libitum) intake was recorded individually. Concentrate (2kg of pre-calver nuts+/-YS for pre-calving and 8 kg of lactating nuts+/-YS for post-calving) feeding was controlled individually. Estimated energy balance PP was calculated on a weekly basis individually as the difference between the net energy (NE) intake and the sum of NE for maintenance and milk production. Insulin and IGF-I concentrations were determined on days 14 and 7 pre-calving and 1, 5, 15, 25 and 35 post-calving. Daily ovarian ultrasonography was performed from day 10 PP to monitor the size and development of the first dominant follicle (>10mm in diameter with absence of other large growing follicles), first ovulatory follicle and days to first ovulation PP. Pre-ovulatory peak of serum oestradiol concentration was determined during the 2 days before ovulation day. Cows with high BCS (over-conditioned) at calving ingested less NE, produced more milk NE output, and consequently had a significantly (P<0.05) exacerbated negative energy balance in comparison with low BCS cows (moderately conditioned) during early lactation. Higher (P<0.05) insulin concentrations and a tendency for higher (P=0.06) pre-ovulatory peak oestradiol concentrations in low BCS group were detected in the early PP period. Supplementing the diet with YS had no effect (P>0.10) on NE intake, NE milk output or energy balance. On the other hand it increased (P<0.01) insulin concentration and tended to increase (P=0.07) pre-ovulatory peak oestradiol concentrations and the size of first ovulatory follicle (P=0.09) early PP. Feeding YS had no effect on energy status of lactating dairy cows with high or low BCS at calving, whilst it improved serum insulin concentration, pre-ovulatory peak of oestradiol and the size of first ovulatory follicle in the early PP period. These observed effects of YS supplementation require to be substantiated with further research.

Abstract: To investigate the effects of pregnancy or post-ovulatory progesterone (P(4)) supplementation on the expression of oestrogen and P(4) receptors (ESRs and PGRs) in the bovine uterus, heifers (n=263) were randomly assigned to the following treatments: i) cyclic, normal P(4); ii) cyclic, high P(4); iii) pregnant, normal P(4); and iv) pregnant, high P(4) on days 5, 7, 13 and 16 of pregnancy/oestrous cycle. Elevated P(4) was achieved through P(4)-releasing intravaginal device insertion on day 3 after oestrus, resulting in increased concentrations from day 3.5 to 8 (P<0.05) in the high groups than in the normal groups. Irrespective of treatment, PGR and ESR1 mRNA expressions were highest on days 5 and 7 and decreased on day 13 (P<0.05), while ESR2 mRNA expression increased on day 7 (P<0.05) and similar levels were maintained within the normal P(4) groups subsequently. Expression in the high P(4) groups decreased on day 13 (P<0.05). PGR-AB and PGR-B protein expressions were high in the luminal and superficial glands on days 5 and 7, but by day 13, expression had declined to very low or undetectable levels and high P(4) concentration tended to decrease or decreased significantly (P<0.05) the expression in these regions on days 5 and 7. ESR1 protein expression was high, with no treatment effect. ESR2 protein was also highly expressed, with no clear effect of treatment. In conclusion, early post-ovulatory P(4) supplementation advances the disappearance of PGR protein from the luminal epithelium on days 5 and 7, and decreases ESR2 mRNA expression during the mid-luteal phase, but has no effect on PGR or ESR1 mRNA expression.

Abstract: The aim of the present study was to compare the hormonal and metabolic characteristics and endometrial gene expression profiles in beef heifers yielding either a viable or degenerate embryo on Day 7 after insemination as a means to explain differences in embryo survival. Oestrus was synchronised in cross-bred beef heifers (n = 145) using a controlled internal drug release (CIDR)-prostaglandin protocol. Heifers (n = 102) detected in standing oestrus (within 24-48 h after CIDR removal) were inseminated 12-18 h after detection of oestrus (Day 0) with frozen-thawed semen from a single ejaculate of a bull with proven fertility. Blood samples were collected from Day 4 to Day 7 after oestrus to measure progesterone (on Days 4, 5 and 7), insulin and insulin-like growth factor (IGF)-I (on Days 4 and 6) and urea (on Day 7) concentrations. All animals were killed on Day 7. Uterine pH was determined at the time of death. Animals from which an embryo was recovered were classified as either having a viable embryo (morula/blastocyst stage; n = 32) or a retarded embryo (arrested at the two- to 16-cell stage; n = 19). In addition, 14 single-celled unfertilised oocytes were recovered, giving an overall recovery rate of 64%. There was no significant difference in the blood parameters determined or uterine pH at the time of death between heifers with either a viable or retarded embryo. The relative abundance of nine transcripts (i.e. MOGAT1, PFKB2, LYZ2, SVS8, UHRF1, PTGES, AGPAT4, DGKA and HGPD) of 53 tested in the endometrial tissue differed between heifers with a viable or retarded embryo. Both LYZ2 and UHRF1 are associated with regulation of the immune system; PFKFB2 is a mediator in glycolysis; MOGAT, AGPAT4 and DGKA belong to the triglyceride synthesis pathway; and PTGES and HGPD belong to the prostaglandin pathway. Both these metabolic pathways are important for early embryonic development. In conclusion, retarded embryo development in the present study was not related to serum progesterone, IGF-I, insulin or urea concentrations, nor to uterine pH at the time of death. However, altered expression of genes involved in the prostaglandin and triglyceride pathways, as well as two genes that are closely associated with the regulation of immunity, in the endometrium may indicate a uterine component in the retardation of embryo development in these beef heifers.

Abstract: Beef suckler farms (194 farms throughout 13 counties) were assessed once with housed cattle and once with cattle at grass using an animal welfare index (AWI). Twenty-three of the 194 farms were revisited a year later and re-evaluated using the AWI and the Tier-Gerechtheits-Index 35L/2000 (TGI35L/2000). Thirty-three indicators were collected in five categories: locomotion (5 indicators); social interactions (between animals) (7), flooring (5), environment (7) and Stockpersonship (9). Three indicators relating to the size of the farm were also collected.Improving animal welfare is an increasingly important aspect of livestock production systems predominantly due to increased consumer concern about the source of animal products. The objectives were (i) to evaluate animal welfare of Irish beef suckler herds using an animal welfare index (AWI), (ii) to examine correlations between parameters, how they influence the AWI and investigate the applicability of the parameters used, (iii) to investigate the impact of the activity of the farmer (full-time or part-time), the interest of the farmer and the number of animals on the AWI.

Abstract: The objective was to investigate the effect of sea transport on the physiological, behavioural and performance responses of bulls. One-hundred and eleven bulls (mean body weight (standard error of the mean) 429 (5.7kg)) were randomly assigned to one of three treatments; control (C; n=54) bulls were housed in 6 pens at Teagasc, Grange Research Centre at a stocking density of (1), 1.7 m(2)/head (C1.7; 3 pens) and (2), 3.4 m(2)/head (C3.4; 3 pens) and (3), transported (T) bulls (n=57) were penned at a space allowance of 1.7m(2)/head (6 pens) and allocated to one of five decks on the shipping vessel. C and T bulls were subjected to the same live weight (d -2), blood sampling and rectal temperature (d -1) measurements pre-transport and on d 3, d 6, d 9 and d 11 of the study. T bulls had greater (P<0.05) live weight gain (+4.4%) compared with C1.7 bulls (-2.0%) and C3.4 (+0.13%)). Time spent lying was greater (P<0.05) among C1.7 and C3.4 bulls (9.9% and 53.3%, respectively) compared with T bulls (45.8%). Rectal body temperature was not different (P>0.05) among treatment groups throughout the study. At d 11, neutrophil % was greater (P<0.05) in transported bulls on decks 1, 2, 4 and 5 compared with C1.7 and C3.4 treatments. Plasma cortisol concentrations were not different (P>0.05) between control and transported bulls. Plasma creatine kinase (CK) activity was lower (P<0.05) among C3.4 and T bulls on decks 2, 3, 4 and 5 compared with d 3 values. In conclusion, the welfare of bulls transported by sea on the sea journey was not adversely affected. Housing control bulls at a reduced space allowance (1.7m(2)) had a negative effect on live weight gain.

Abstract: The hypothesis was that supplementation during the late prepartum period will differentially affect reproductive and productive variables according to parity. Primiparous (n=22) and multiparous (n=22) pregnant autumn calving Holstein cows were stratified in two groups according to parity (primiparous or multiparous) and within each group were randomly assigned to two treatments: (a) low supplemented (LS) or (b) high supplemented (HS) prepartum diet. The LS group was offered 5.2 kg/cow/day (DM basis) of wheat silage, and the HS group 4.7 kg cow/day (DM basis)/of corn silage and 3.6 kg (DM basis) of wheat bran+12 g of urea. Both groups grazed on natural pastures. After calving, all cows received the same diet. The experimental period was from 3 weeks before calving to 7 weeks postpartum (PP); body condition score (BCS) and blood samples for hormonal analyses were obtained weekly and ovarian ultrasonography was conducted three times per week. The loss in BCS around calving was less pronounced in HS cows, but only multiparous supplemented cows maintained BCS throughout the study. Non-esterified fatty acids (NEFA) increased during the prepartum period in the LS but not in the HS cows, with peak values occurring on day 14 PP in all groups. During the remainder of the experiment NEFA was greater in LS than in HS cows. Prepartum treatment did not affect the proportion of cows that had ovulations from the first dominant follicle postpartum, but decreased the interval to first ovulation in multiparous cows (22.9 compared with 38.2 days; P<0.05). This was associated with greater plasma IGF-I concentrations at the time the dominant follicle of the first follicular wave reached its maximum diameter (8.0 compared with 3.6 nmol/L; P<0.05). However, prepartum treatment had no effect on onset of ovarian activity in primiparous cows. Supplementation had no effect on milk production or milk protein percentage but increased milk fat percentage. We conclude that feeding a high-supplemented prepartum diet to multiparous cows allowed them to maintain BCS around calving, and this was associated with greater concentrations of IGF-I and an earlier onset of estrous cycles after calving.

Abstract: The objectives were to determine the effects of (i) time during the first FSH increase of the estrous cycle (time-course study) and (ii) exogenous steroid treatment (steroid feedback study) on the relationship between circulating serum gonadotropins, and the proportions of pituitary cells immunoreactive for gonadotropins and steroid receptors during the estrous cycle in heifers. Pituitaries were collected from heifers (n=40) slaughtered at 13h (n=8), 30h (n=24) and 66h (n=8) after estrous onset, corresponding to before, during and after the first FSH increase of the estrous cycle. Heifers slaughtered during the FSH increase (at 30h) either received no treatment (n=8), or were treated (n=16) with estradiol benzoate and/or progesterone before slaughter. During the time-course study, the proportion of pituitary cells immunoreactive for FSH increased (P<0.05) during the first transient FSH increase reflecting serum concentrations. The proportion of pituitary cells immunoreactive for LH was unaltered, a reflection of serum LH concentrations. The proportion of estrogen receptors (ER)-alpha, but not ER-beta, was decreased (P<0.05) at 30h compared with at either 13 or 66h. During the steroid feedback study, exogenous progesterone with or without estradiol suppressed (P<0.05) the proportions of pituitary cells immunoreactive for gonadotropins, serum FSH concentrations and LH pulse frequency. Steroid treatment did not alter the proportion of pituitary cells positive for estrogen receptors (alpha and beta). While progesterone receptors (PR) were not detected in the anterior pituitary by immunohistochemistry during the early estrous cycle or in response to steroid treatment, quantitative real-time PCR revealed that mRNA for progesterone receptors was expressed at very low levels. The expression of pituitary PR mRNA was decreased (P<0.05) at 30 and 66h compared with 13h, and was suppressed (P<0.05) following steroid treatments. Alterations in pituitary steroid receptors are implicated in the differential regulation of gonadotropin secretion during the first transient FSH rise, but not in response to exogenous steroids. The time-course study and steroid feedback responses support the hypothesis that LH pulse frequency is tightly linked to regulation of GnRH pulse frequency. Serum FSH is regulated by its own synthesis, as reflected by pituitary FSH content and perhaps by alterations in pituitary sensitivity to circulating steroids by changes in steroid receptor content.

Abstract:

Abstract: Progesterone is essential for establishment and maintenance of pregnancy in mammals. The objective of this study was to examine the effect of elevating progesterone during the different physiological stages of early embryo development on embryo survival. Estrus was synchronized in cross-bred beef heifers (n=197, approximately 2-years old) and they were inseminated 12-18h after estrus onset (=Day 0). Inseminated heifers were randomly assigned to 1 of 3 treatments: (1) Control, n=69; (2) progesterone supplementation using a Controlled Internal Drug Release Device (CIDR) from Day 3 to 6.5, n=64; or (3) progesterone supplementation using a CIDR from Day 4.5 to 8, n=64. Body condition (BCS) and locomotion scores (scale of 1-5) were recorded for all animals. Animals with a locomotion score >/=4 (very lame) were excluded. Embryo survival rate was determined at slaughter on Day 25. Conceptus length and weight were recorded and the corpus luteum (CL) of all pregnant animals was dissected and weighed. Supplementation with exogenous progesterone increased (P<0.05) peripheral progesterone concentrations, but did not affect embryo survival rate compared with controls. Mean CL weight, conceptus length and conceptus weight were not different between treatments. There was a positive relationship (P<0.04) between the increase in progesterone concentrations from Days 3 to 6.5 and embryo survival rate in treated heifers and a similar trend existed between the increase from Days 4.5 to 8 (P<0.06). There was also a positive relationship (P<0.05) between the progesterone concentration on Day 6.5 and the embryo survival rate in treated heifers. A direct correlation was seen between locomotion score and embryo survival rate, with higher (P<0.05) early embryo survival rates in heifers with a lower locomotion score. In conclusion, supplementation with progesterone at different stages of early embryo development increased peripheral progesterone concentration and resulted in a positive association between changes in progesterone concentration during the early luteal phase and embryo survival rate. Supplementation with progesterone had no effect on either CL weight or conceptus size in pregnant animals. Lameness had a significant negative effect on early embryo survival.

Abstract: Castration of male cattle has been shown to elicit inflammatory reactions and acute inflammation is initiated and sustained by the participation of cytokines.

Abstract: Two experiments were designed to evaluate models for generation of low circulating progesterone concentrations during early pregnancy in cattle. In Experiment 1, 17 crossbred heifers (Bos taurus) were assigned to either prostaglandin F(2alpha) (PGF(2alpha)) administration on Days 3, 3.5, and 4 (PG3; n=9) or to control (n=8). Blood samples were collected from heifers from Days 1 to 9 for progesterone assay. Progesterone concentrations were decreased (P<0.03) between 18 and 48h after first PGF(2alpha) treatment in heifers assigned to PG3 compared with that of controls. In Experiment 2, 39 crossbred heifers detected in estrus were inseminated (Day 0) and assigned to either (1) PGF(2alpha) administration on Days 3, 3.5, and 4 (PG3; n=10), (2) PGF(2alpha) administration on Days 3, 3.5, 4, and 4.5 (PG4; n=10), (3) Progesterone Releasing Intravaginal Device (PRID) insertion on Day 4.5 with PGF(2alpha) administration on Days 5 and 6 (PRID+PGF(2alpha); n=10), or (4) control (n=9). Blood samples were collected daily until Day 15, and conceptus survival rate was determined at slaughter on Day 16. Progesterone concentrations during the sampling period in the PG3 and PG4 groups did not differ but were less than that of controls (P<0.01). After an initial peak, progesterone concentrations in the PRID+PGF(2alpha) group were similar to that of controls. More heifers in the PG4 group (6 of 10) had complete luteal regression than did those in the PG3 group (3 of 10). Conceptus survival rate on Day 16 did not differ between groups. There was a significant correlation between progesterone concentration on Days 5 and 6 and conceptus size on Day 16. In summary, treatment with PGF(2alpha) on Days 3, 3.5, and 4 postestrus appeared to provide the best model to induce reduced circulating progesterone concentrations during the early luteal phase in cattle.

Abstract: The postovulatory rise in circulating progesterone (P4) concentrations is associated with increased pregnancy success in beef and dairy cattle. Our study objective was to determine how elevated P4 alters endometrial gene expression to advance conceptus development. Synchronized heifers were inseminated (Day 0) and randomly assigned to pregnant high P4 or to pregnant normal P4. All high P4 groups received a P4-release intravaginal device on Day 3 after insemination that increased P4 concentrations up to Day 7 (P < 0.05). Tissue was collected on Day 5, 7, 13, or 16 of pregnancy, and endometrial gene expression was analyzed using the bovine Affymetrix (Santa Clara, CA) microarrays. Microarray analyses demonstrated that the largest number of P4-regulated genes coincided with the day when the P4 profiles were different for the longest period. Genes with the largest fold change increase (such as DGAT2 and MSTN [also known as GDF8]) were associated with triglyceride synthesis and glucose transport, which can be utilized as an energy source for the developing embryo. Temporal changes occurred at different stages of early pregnancy, with the greatest difference occurring between well-separated stages of conceptus development. Validation of a number of genes by quantitative real-time PCR indicated that P4 supplementation advances endometrial gene expression by altering the time (FABP, DGAT2, and MSTN) or duration (CRYGS) of expression pattern for genes that contribute to the composition of histotroph.

Abstract: The objective was to investigate measures of neutrophil function in response to banding or burdizzo castration of bulls. Thirty-two Holstein-Friesian bulls (14 mo old, 505 +/- 7.8 kg of BW) were assigned to 1 of 4 treatment groups: 1) sham-handled control (CON); 2) banding castration alone (BAND); 3) burdizzo castration alone (BURD); or 4) cortisol infusion (CORT) as a further control group. For each group on d -14, 8 animals (2 animals/treatment) were tied up in tie stalls (day of treatment = d 0). At -2, 2, 6, 12, 24, 48, 72, and 144 h relative to treatment time, blood samples were collected for analyses of neutrophil phagocytosis and respiratory burst, neutrophil CD62-L expression, and serum IL-8 concentration. Leukocyte counts, phagocytosis activity, and CD62-L expression were similar (P > 0.05) among the 4 treatment groups. The BURD castrates had greater burst activity compared with BAND castrates (P = 0.048) and CON (P = 0.01) at 72 h posttreatment. The BURD castrates had a greater percentage of granulocyte positive leukocytes (Gr%; P < 0.01) at 2 h posttreatment compared with CON and CORT bulls. The BURD castrates had greater (P < 0.05) Gr% compared with BAND, CON, and CORT animals at 24, 48, and 72 h posttreatment. The BURD and BAND castrates had greater Gr% (P < 0.05) compared with CORT bulls at 144 h posttreatment. In general, BAND, BURD, and CORT did not affect serum IL-8 concentration. Banding castration, BURD, and CORT did not induce leukocytosis, whereas BURD induced a modest neutrophilia. Neutrophil functioning in terms of phagocytosis and respiratory burst and serum IL-8 concentration were not compromised by BAND, BURD, and CORT. These findings indicate nonsurgical castration is unlikely to induce a severe acute systemic inflammatory response in terms of neutrophil function.

Abstract: Transportation causes stress in cattle that may alter numerous physiological variables with a negative impact on production and health. The objectives of the current study were to investigate the physiological effects of truck transportation and to characterize a pattern of phenotypes in the circulation that may aid in the early identification of stress-susceptible animals that often succumb to severe respiratory disease. Thirty-six young beef bulls (Aberdeen Angus, n = 12; Friesian, n = 12; and Belgian Blue x Friesian, n = 12) were subjected to a 9-h truck transportation by road. Blood (10 mL) was collected at -24, 0, 4.5, 9.75, 14.25, 24, and 48 h relative to the initiation of transportation (0 h). Plasma was collected for the assay of various metabolic, inflammatory, and steroid variables, and total leukocyte counts were determined in whole blood at each time point. Body weight and rectal temperature were recorded at -24, 9.75, and 48 h. Transportation decreased measures of protein metabolism in the plasma, including albumin (P = 0.002), globulin (P < 0.001), urea (P = 0.006), and total protein (P < 0.001), and increased creatine kinase (P < 0.001). The energy substrate beta-hydroxybutyrate was not changed (P = 0.27). Acute phase proteins haptoglobin and fibrinogen were both decreased (P < 0.001), whereas total leukocyte counts were elevated (P = 0.002). Circulating steroid concentrations were altered, because a classical acute increase in plasma cortisol was observed with the onset of transit (P < 0.001), in association with a decrease in dehydroepiandrosterone (P = 0.07), resulting in a profound increase in cortisol:dehydroepiandrosterone ratio (P < 0.001). Plasma testosterone was decreased, whereas plasma progesterone was increased (P < 0.001) in association with the increase in cortisol (P < 0.001). There was also an effect of breed for all variables except plasma urea, creatine kinase, and testosterone, perhaps indicating that a genetic component contributed to the physiological response to transportation stress, although without any clear trend. Taken together, this profile of physiological variables in the circulation of transportation-stressed bulls may aid in the future detection of disease-susceptible cattle after transportation. Further research to validate these potential biomarkers is necessary.

Abstract: Higher systemic progesterone in the immediate post-conception period is associated with an increase in embryonic growth rate, interferon-tau production and pregnancy rate in cattle. The objective of this study was to examine the effect of increasing progesterone concentration on Day 3 on subsequent embryo survival and development. Oestrus (Day 0) was synchronised in beef-cross heifers (n=210) and approximately two-thirds of the heifers were inseminated with semen from a proven sire, while the remainder were not inseminated. In order to produce animals with divergent progesterone concentrations, half of the animals received a progesterone-releasing intravaginal device (PRID) on Day 3 of the oestrous cycle, which was left in situ until slaughter. The four treatment groups were: (i) pregnant, high progesterone; (ii) pregnant, normal progesterone; (iii) non-pregnant, high progesterone; and (iv) non-pregnant, normal progesterone. Animals were blood-sampled twice daily from Days 0 to 8 and once daily thereafter until slaughter on Days 5, 7, 13 or 16, corresponding to the 16-cell stage, the blastocyst stage, the beginning of elongation and the day of maternal recognition of pregnancy, respectively. Embryos were recovered by flushing the tract with phosphate-buffered saline and characterised by stage of development and, in the case of Days 13 and 16, measured. Data were analysed by mixed models ANOVA, Chi-square analysis and Student's t-test where appropriate. Insertion of a PRID on Day 3 increased (P<0.05) progesterone concentrations from Day 3.5 onwards. There was no difference between treatments in the proportion of embryos at the expected stage of development on Days 5 or 7 (P>0.05). While not significantly different, the proportion of viable embryos recovered was numerically greater in the high progesterone group on both Day 13 (58 v. 43%) and Day 16 (90 v. 50%). Elevation of progesterone significantly increased embryonic length on Day 13 (2.24+/-0.51 mm v. 1.15+/-0.16 mm, P=0.034) and Day 16 (14.06+/-1.18 cm v. 5.97+/-1.18 cm, P=0.012). In conclusion, insertion of a PRID on Day 3 of the oestrous cycle increased serum progesterone concentrations on subsequent days, which, while having no phenotypic effect on embryonic development on Days 5 or 7, was associated with an increase in embryonic size on Days 13 and 16.

Abstract: Stress and its association with altered immune function and incidence of respiratory diseases in cattle have lead to concerns over animal health and welfare during truck transportation. Previously, bulls subjected to transportation stress displayed altered expression of candidate neutrophil genes, warranting a broader investigation of the neutrophil transcriptome and possible associations with fluctuations in circulating steroid hormones. In the current study, blood was collected from six Belgian BluexFriesian bulls at -24, 0, 4.5, 9.75, 14.25, 24, and 48h relative to initiation of 9h of truck transportation. Plasma concentrations of cortisol, dehydroepiandrosterone (DHEA), progesterone, and testosterone were measured; cortisol:DHEA ratios were computed. Neutrophil gene expression was monitored by microarray analysis using bovine immunobiology (BOTL-5) microarrays. Eighty-eight genes were identified as being differentially expressed at P<0.05. Putatively affected genes were grouped into ontological clusters; those of greatest interest for qRT-PCR validation were involved in immune response, apoptosis, wound healing, and several of currently unknown function. Confirmed gene expression changes supported the dramatic effects of transportation stress on the bovine neutrophil transcriptome. Temporal correlations between gene expression profiles and circulating total leukocyte and neutrophil counts were apparent. However, few relationships between gene expression and plasma steroid profiles were detected, possibly due to the biological time-lag between these variables not captured by the blood collection schedule. Further investigation into the factors underlying neutrophil gene expression changes and validations at the protein and cell behavior levels will lead to a better understanding of altered innate immunity in cattle during transportation stress.

Abstract: Oestrous synchronisation is an important strategy to improve reproductive management of cattle. The use of oestradiol-17beta, and its related ester derivatives, in food-producing cattle for the purposes of oestrous synchronisation is prohibited in the European Union since October 2006; a serious limitation in the implementation of large-scale use of cost effective synchronisation regimens in both dairy and beef herds. This has obvious consequences within the EU and also in other countries that have restricted the use of oestradiol following the EU ban. Oestrous synchronisation is an important facilitator for the use of artificial insemination, a necessary part of any national herd genetic improvement scheme. Presently, only 35% of the Irish dairy herd is bred by artificial insemination; and facilitation rather than restriction is required to increase this percentage. Ideally synchronisation of oestrus should increase submission rates, improve or at least not affect conception rates, and thus, increase overall pregnancy rate at the end of the breeding season. This should reduce the proportion of cows to be culled. This paper aims to review the oestrous synchrony options available in EU countries and other countries affected by the European ban on oestrogenic compounds being used for oestrous synchrony protocols. Currently, the options available for oestrous synchronisation are generally not as effective, efficient or cost effective as those that incorporated use of oestrogenic compounds.

Abstract: There is a variable anoestrous period following parturition in the cow. Follicular growth generally resumes within 7-10 days in the majority of cows associated with a transient follicle-stimulating hormone (FSH) rise that occurs within 3-5 days of parturition. Dairy cows that are not nutritionally stressed generally ovulate their first post-partum dominant follicle (approximately 15 days), whereas beef suckler cows in good body condition normally have a mean of 3.2 +/- 0.2 dominant follicles (approximately 30 days) to first ovulation; and beef cows in poor body condition have a mean of 10.6 +/- 1.2 dominant follicles (approximately 70-100 days) to first ovulation. The lack of ovulation of dominant follicles during the post-partum period is associated with infrequent luteinizing hormone (LH) pulses, with both suckling and low level of nutrition being implicated in the prolonged suppression of LH pulses in the absence of progesterone. In dairy cows, the normal pattern of early resumption of ovulation may be delayed in high-yielding Holstein-type cows generally because of the effects of severe negative energy balance, dystocia, retained placental membranes and uterine infections. First ovulation in both dairy and beef cows is generally silent (i.e., no behavioural oestrus) and is generally (>70%) followed by a short cycle. The key to optimizing resumption of ovulation in both beef and dairy cows is appropriate pre-calving nutrition and management so that cows calve down in optimal body condition (body condition score; BCS; 2.75-3.0) with post-partum body condition loss restricted to <0.5 BCS units.

Abstract: The effect of various space allowances on the pituitary, adrenal and immune responses and on performance was investigated in 72 mature Holstein x Friesian beef bulls. The animals (weighing 403+/-3.5 kg) were blocked by weight and randomly assigned to two groups (familiar, F, and unfamiliar, UF) x three treatments (1.2, 2.7 and 4.2m(2) per bull; n=24 per space allowance), and housed for 83 days in 18 pens (n=4 per pen). Blood samples were collected on days -1, 0, 3, 14, 36 and 77 with respect to mixing and housing on day 0. The bulls were given exogenous adrenocorticotrophic hormone (ACTH) on day 3 and corticotrophin-releasing hormone (CRH) on days 14, 36 and 77. Basal plasma cortisol concentration was not affected (P>0.05) by mixing F and UF bulls. On day 3, basal cortisol was greater (P<0.05) in bulls housed at 1.2 than those at 2.7 and 4.2m(2) space allowances while no effect was observed in ACTH-induced plasma cortisol concentration among treatments. Following CRH administration, there was no effect (P>0.05) of treatment and treatment x time on plasma ACTH concentration. On day 14, interferon-gamma production was lower (P<0.05) in the bulls housed at 4.2 vs. 2.7 m(2) and was intermediate but not significantly different (P>0.05) for those housed at 1.2m(2). Animals housed at either space allowances had significant (P<0.05) neutrophilia, lymphopenia, eosinopenia and decreased haemoglobin on day 3 compared with day 0. The liveweight gain from days 0 to 83 was lower (P<0.05) in bulls housed at 1.2 compared with those at 2.7 and 4.2m(2). Housing bulls at 1.2m(2) space allowance had a detrimental effect on their growth and was associated with an acute rise in plasma cortisol concentration (on day 3) compared with those having space allowances of 2.7 and 4.2m(2)/bull.

Abstract: The effects of transporting Holstein Friesian bulls (n=72; bodyweight 403+/-3.5 kg) for 12h by road were examined. Adrenal, haematological and immune responses, body temperature and performance were recorded. The animals had been previously housed for 96 days at three space allowances (1.2, 2.7 or 4.2m(2) per bull). The bulls were allocated to one of two treatments: T (transport for 12h; n=16 per space allowance) and C (control; n=8 per space allowance). Basal cortisol plasma concentrations and interferon (IFN)-gamma production from cultured lymphocytes did not show any statistically significant difference (P>0.05) following the housing period. Removing bulls from their home pens and walking them to the pre-loading crush facility, loading onto the transporter, and unloading following the 12h road journey, significantly (P<0.001) increased plasma cortisol concentration. The bulls housed at 4.2m(2) had greater (P<0.05) plasma cortisol concentrations than bulls housed at 1.2m(2) at loading, unloading, or on return to the crush holding facility; those housed at 1.2m(2) had greater (P<0.05) plasma cortisol concentrations than bulls housed at 2.7 and 4.2m(2) in their home pens after transport. There was an increased (P<0.05) plasma cortisol response in the T than in the C bulls following adrenocorticotrophic hormone administration. Transport significantly reduced (P<0.05) IFN-gamma production, lymphocyte % and body weight and significantly increased (P<0.05) neutrophils, eosinophils, packed cell volume, red blood cell numbers and haemoglobin. In conclusion, housing bulls for 96 days in a range of space allowances did not affect basal cortisol response or immune function parameters. Whereas transport increased plasma cortisol and reduced the immune response in the short-term, the changes were transient and within normal physiological ranges, suggesting that 12h road transport had no adverse effect on welfare status over the longer term. Furthermore, transport of bulls housed at increased space allowance (4.2m(2)/bull) resulted in a greater plasma cortisol response, albeit still within normal physiological range.

Abstract: Dominant follicles are those that continue to develop and have the potential to ovulate while subordinate follicles regress. Characteristics of dominant follicles include a larger diameter, higher intrafollicular estradiol, and lower IGF-binding protein (IGFBP)-4 concentrations compared with other cohort follicles. Follicle development is regulated by endocrine hormones that act via intracellular signaling pathways. Here, we show the differences in Akt, Erk, c-Jun N-terminal protein kinase, and p-38 signaling pathways between dominant and subordinate follicles at the dominance stage of the follicle wave. However, earlier in the follicle wave (dominant follicle selection), there were only differences in the levels of Akt and Erk signal transduction proteins among dominant and subordinate follicles. Using this profile of Akt and Erk protein expression in granulosa and theca cells of selected dominant follicles compared with subordinate follicles, we suggest a predictive model to identify future dominant and subordinate follicles from the pool of otherwise similar cohort follicles at the time of follicle wave emergence. We conclude that the Erk and Akt signal transduction pathways are important for dominant follicle selection and development and, furthermore, that the observed differences in these pathways mark the future dominant follicle from subordinate follicles before differences in follicular diameter, follicular fluid estradiol, and IGFBP-4 concentrations are apparent.

Abstract: The transportation of beef cattle results in a stress response that is associated with increased susceptibility and severity of respiratory diseases, presumably due to an alteration in immune function. Neutrophils are phagocytic immune cells important in lung defense and are also targets of the stress response. The objective of this study was to determine if a 9h transportation of young bulls by road induced changes in the expression of candidate genes known to be important in neutrophil-mediated defense and inflammation in the lung. These neutrophil genes encompassed functions of apoptosis (A1 and Fas), tissue remodeling (MMP-9), vascular margination (L-selectin), bacterial killing (BPI), and wound healing (betaglycan), as well as responsiveness of the cells to stress-induced increases in glucocorticoid hormones (GRalpha). To explore gene expression changes, blood was collected, plasma harvested, and neutrophils isolated from six Belgian Blue x Friesian bulls (231+/-7.0 kg in weight; 282+/-4 days of age) at -24, 0, 4.5, 9.75, 14.25, 24, and 48h relative to commencement of a 9h road transportation by truck. Plasma cortisol concentrations were elevated at 4.5 and 9.75h, peaking at 50.64+/-4.46 ng/mL (P<0.0001) and confirming that the animals experienced stress. Blood neutrophil count was elevated between 4.5 and 14.25h (P<0.0001), reaching a peak that was over 3-fold higher than the -24h concentration. Neutrophil Fas gene expression was acutely down-regulated (P=0.02) by transportation stress, while expressions of MMP-9, l-selectin, and BPI were profoundly up-regulated (P=0.003, 0.002, and <0.001 respectively). However, no changes in neutrophil expressions of betaglycan, GRalpha, and A1 were detected. It is concluded that a 9h transportation of young bulls induces a gene expression signature in blood neutrophils that increases their circulating numbers and may enhance their pro-inflammatory and anti-bacterial potential.

Abstract: The objective was to determine the pattern of IGFBP-2, -3 and -4 gene expression and follicular fluid concentrations of IGFBP-2, -3, -4 and -5 during emergence, selection and dominance of the first follicle wave of the estrous cycle in cattle and during exogenous steroid treatment. Heifers (n = 35) were ovariectomized at 36 (n = 7), 66 (n = 8), 84 (n = 12) and 108 (n = 8) h after the onset of estrus. Heifers in the 84 h ovariectomy group were sub-divided to receive either no treatment (n = 6) or were treated with a progesterone-releasing intravaginal device (n = 6, PRID) and 0.75 mg estradiol benzoate i.m. at the approximate time of ovulation, 30 h post estrus until ovariectomy. Within heifers the four largest follicles recovered following ovariectomy were ranked on size (F1, F2, F3 and F4). At 36 h IGFBP gene expression and follicular fluid IGFBP concentrations were similar in all follicles (F1-F4). Mean diameter of the F1 follicle increased (P < 0.05) between 36 and 84 h with no difference between 84 and 108 h. The F1 follicle had the highest (P < 0.05) concentration of estradiol compared with the F2, F3 and F4 at 84 and 108 h. There was no granulosa cell IGFBP-2 mRNA in F1 follicles at 84 or 108 h. Intrafolliclar IGFBP-2 concentrations were lower (P < 0.05) in the F1 compared with F3 and F4 follicles at 108 h. There was no difference in theca cell IGFBP-4 mRNA expression at 108h, but amounts of follicular fluid IGFBP-4 were lower (P < 0.05) in F1 follicles compared with F3 and F4 follicles at 108 h. IGFBP-3 mRNA was localized in the theca layer of all follicles examined with no difference in expression or follicular fluid concentrations during emergence, selection and dominance of the first follicle wave. IGFBP-5 concentrations were higher (P < 0.05) in follicular fluid of F3 follicles at 108 h compared with the F3 at 36 h. In conclusion follicular dominance was associated with low or decreased follicular fluid concentrations of IGFBP-4 and -5, increased estradiol and differential regulation of IGFBP production.

Abstract: The aim of this study was to evaluate the effect of active immunization against GnRH on ovarian activity, plasma progesterone and estradiol concentrations and on estrous behavior in adult mares. Eighteen cyclic mares were randomly divided into a treatment and control group. Nine mares were immunized twice with 2 mL (400 microg GnRH-protein conjugate) of a GnRH-vaccine (Improvac, CSL Limited, Australia) administered intramuscularly, 4 weeks apart. Control mares received the same amount of saline solution. Ovaries and uterus of all mares were examined weekly by ultrasonography from 3 weeks before to 60 weeks after first immunization. Thereafter, vaccinated mares were evaluated monthly until 100 weeks after first vaccination. In addition, mares were teased with a stallion for assessment of estrous behavior and blood was collected for progesterone, estradiol-17beta and GnRH antibody titer determination. Results demonstrate that vaccination against GnRH significantly (P<0.05) influenced all parameters, except estradiol-17beta concentration. All vaccinated mares ceased reproductive cyclicity (plasma progesterone <1 ng/mL, follicles <3 cm) within 8 weeks after the first injection and ovarian activity remained suppressed for a minimum of 23 weeks. Five mares resumed cyclicity (follicles >3 cm, progesterone >1 ng/mL) while three mares showed only follicular activity (follicles >3 cm) and one mare remained completely suppressed for the entire duration of the study. In spite of ovarian suppression, four mares expressed sporadic and one mare continuous estrous behavior. In conclusion, reproductive cyclicity in adult mares can be successfully suppressed by immunization against GnRH but the timing of resumption of cyclicity is highly variable and estrous behavior may occur in spite of ovarian suppression.

Abstract: The objective of this study was to determine the effect of carprofen (C) administration before banding or burdizzo castration of bulls on cortisol, in vitro interferon-gamma (IFN-gamma) production, acute-phase proteins, feed intake, and growth. Fifty Holstein Friesian bulls (5.5 mo old; 191 +/- 3.7 kg) were blocked by weight and assigned randomly to 1 of 5 treatments (n = 10/treatment): 1) untreated control (2) banding castration at 0 min (Band); 3) Band following an i.v. injection of 1.4 mg/kg of BW of C at -20 min (Band+C); 4) Burdizzo castration at 0 min (Burd); or 5) Burd following 1.4 mg/kg of BW of C at -20 min (Burd+C). Castration acutely increased plasma cortisol concentrations compared with control; no significant differences occurred in peak and interval to peak cortisol responses between Band and Band+C or Burd and Burd+C groups. The administration of C in Band+C reduced (P < 0.05) the cortisol concentration between 6 and 12 h postcastration compared with Band animals. Overall, the integrated cortisol response was greater (P < 0.05) in the castrates than in control, whereas C treatments tended to reduce this response compared with Band (P = 0.08) and Burd (P = 0.07), respectively. Plasma fibrinogen was elevated in Band animals on d 14 and in Burd animals on d 3 and 14. Carprofen administration reduced Band- and Burd-induced fibrinogen production on d 14 and 3, respectively. Plasma haptoglobin was elevated in Band animals on d 3 and 35 compared with control, and C administration was effective in reducing the haptoglobin elevation on d 35 in Band+C compared with Band. There were no differences among treatments in in vitro IFN-gamma production induced by concanavalin A and phytohemagglutinin on d 1 and 2. Overall from d -1 to 16, there were no DMI differences among treatments. From d -1 to 35, there were no ADG differences among treatments. In conclusion, banding and burdizzo castration increased plasma cortisol with no change in in vitro IFN-gamma production. Carprofen (1.4 mg/kg of BW) tended to reduce the integrated cortisol response, and it reduced cortisol secretion in banded animals between 6 and 12 h postcastration. There was an increased acute-phase protein production following castration; this response was effectively moderated by the administration of C before castration.

Abstract: The objective was to determine if the endocrine status of the animal dictates the responsiveness of gonadotrophs to estradiol, activin, inhibin and follistatin; hormones implicated in the differential release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Bovine pituitaries were obtained at 13 (n=8), 30 (n=24) and 66 (n=8) h after the onset of estrus, corresponding to before, during and the end of the first FSH increase of the estrous cycle which follows the pre-ovulatory gonadotropin surge in heifers. Heifers slaughtered at 30 h received no treatment, or were treated with progesterone with or without estradiol before slaughter to suppress the first transient FSH increase. Secretion of FSH from cultured pituitary cells, reflecting the prior in vivo status, was greater (P<0.01) at 30 h than 13 or 66 h, whereas, LH secretion was less (P<0.01) at 13 h compared with 30 h. Treatment with exogenous steroids decreased (P<0.05) the pituitary gland's ability to subsequently secrete FSH and LH. Inhibin and, to a greater extent, estradiol decreased (P<0.01) mean FSH secretion but increased (P<0.05) mean LH secretion. These findings suggest that estradiol and inhibin both have the ability to differentially modulate basal gonadotropin secretion during the first FSH increase of the bovine estrous cycle. Differential regulation of LH and FSH is mediated via an alteration in gonadotropin biosynthesis and basal secretion. Furthermore, the secretory capability of cultured pituitary cells and basal gonadotropin secretion reflect the prior endocrine status of the animal from which pituitaries were obtained.

Abstract: The objective was to determine if a single measurement of plasma insulin-like growth factor-1 (IGF-1) could predict the number of viable embryos obtained from donors and the likelihood of pregnancy in recipients in multiple ovulation and embryo transfer (MOET) programs in cattle. The embryo yields from 101 embryo recoveries were examined in maiden Holstein heifers (n=75) and multiparous Holstein cows (lactating cows n=20, dry cows n=6). Donors were super stimulated with FSH and embryo recovery was done non-surgically 7 days after artificial insemination. Embryos were classified according to the IETS criteria. Pregnancy rates in 100 maiden Holstein heifer recipients were analysed. Recipients were on day 7+/-1 of the estrous cycle at transfer. Pregnancy diagnosis was carried out at day 30 (PD 30) and rechecked at day 60 (PD 60) after transfer. Blood samples from coccygeal vessels taken at the time of embryo recovery (donors) and transfer (recipients) were analysed for IGF-1, insulin, beta-hydroxybutyrate (beta-OHB), non-esterified fatty acids (NEFA), urea and cholesterol. There was a negative correlation between the number of viable embryos and insulin (r=-0.33, P=0.025) in donor heifers. In donor cows, the number of viable embryos was correlated with IGF-1 (r=0.43, P=0.028) and cholesterol (r=-0.43, P=0.027). In recipients, PD30 and PD 60 were not affected by any of the circulating parameters analysed. Insulin, IGF-1 and cholesterol only explained 8.9, 13.9 and 15.8% of the variation in the production of viable embryos, respectively. Several factors affect MOET programs and under the circumstances of the present study the usefulness of hormonal and metabolic profiles as predictors of the outcome of this biotechnology was limited.

Abstract: To investigate the effect of repeated regrouping and repenning (R&R) on the hypothalamic-pituitary-adrenal axis, immune function, blood biochemical and hematological variables, and ADG, 72 Holstein-Friesian (14-mo-old; 441 +/- 3.2 kg) steers were assigned to either the control (C; n = 30) or regrouped (R; n = 42) treatments and housed six per pen in 12 pens. The R steers were exposed to six R&R over 84 d. New pen cohorts were allowed to stabilize for 14 d, and none of the R steers was allowed to share the same pen or penmates where or with whom they were previously housed. Control steers were housed in the same pen with the same penmates. Steers were blood sampled 2 h before and 2 h after the first, third, and sixth R&R. Steers were weighed the day before each R&R. Median area under the plasma cortisol curve (AUC) was greater (P < 0.05) in R than C steers after the first R&R. Following the first, third, and sixth R&R, the median ACTH AUC did not differ between the treatments. Cortisol AUC in R steers decreased (P < 0.001) following the third and sixth compared with the first R&R, however, cortisol AUC in response to exogenous ACTH (following administration of dexamethasone at -12 h) after the third R&R was greater in C than R steers (P < 0.05). Corticotropin-releasing hormone-induced cortisol and ACTH AUC were not different in C vs. R after the sixth R&R. There were no differences among treatments in haptoglobin, fibrinogen, and concanavalin A-induced interferon-gamma after the first, third, and sixth R&R. Albumin, urea, and NEFA were greater (P < 0.05) in R than C steers after the first R&R. beta-Hydroxy-butyrate and glucose concentrations were greater (P < 0.05) in R than C, whereas no changes in the protein and globulin concentrations were found in C vs. R after the sixth R& R. White blood cell, differential and total count, red blood cell, and platelet numbers did not differ in C vs. R after the first and third R&R. Lymphocyte numbers and mean corpuscular volume were greater (P < 0.05) in R than C steers after the sixth R&R. Monocyte numbers were greater (P < 0.05) in R than C steers following first R&R. There was no difference in the overall ADG in C vs. R; however, there was a tendency (P = 0.10) for lesser ADG by R than C steers following second R& R. In conclusion, steers exposed to R&R responded with increased plasma cortisol, albumin, urea, and NEFA. Repeated R&R did not have a sustained detrimental effect on immune and production measurements.

Abstract: This study tested the hypotheses that: (1) either acute stress induced by Burdizzo castration, or cortisol infusion would modulate plasma glucose, insulin and growth hormone (GH) concentrations; and (2) immune modulation induced by cortisol would be dependent on the pattern, intensity and duration of circulating cortisol concentrations. Fifty 9.2-month-old Holstein x Friesian bulls (232 +/- 2.0 kg) were blocked by weight and randomly assigned to one of five treatments (n = 10 per treatment): (1) sham handled control; (2) Burdizzo castration; (3) hydrocortisone infusion to mimic the castration-induced secretion pattern of cortisol; (4) hourly pulse infusion of hydrocortisone; and (5) sustained infusion of hydrocortisone for 8h. Blood samples were collected intensively on day 0, and weekly from days 1 to 35. Castration acutely increased plasma cortisol, GH and haptoglobin concentrations, suppressed lymphocyte in vitro interferon-gamma (IFN-gamma) production, but had no effect on plasma glucose and insulin concentrations. Cortisol infusion to simulate the castration-induced secretion pattern of cortisol, and pulse infusion of cortisol did not suppress the IFN-gamma production. A sustained infusion of cortisol resulted in the transient suppression of IFN-gamma production. Moreover, the sustained cortisol infusion resulted in increased plasma glucose, insulin and GH concentrations. The overall 14-day feed intakes and 35-day growth rates were not affected by treatments. In conclusion, cortisol infusion to induce immune suppression in vivo occurred only at pharmacological doses. Within physiological ranges, cortisol was not associated with the suppression of immune function, indicating that during castration cortisol per se is not responsible for the suppression of in vitro IFN-gamma production.

Abstract: In post-partum anestrous beef cows suckling calves, neither the choice of hormonal regime to ensure the presence of a healthy dominant follicle at the end of a progestagen treatment nor the optimum hormone to induce estrus and ovulation is clear. Twenty-eight beef cows, in good body condition, 25-30 days post-partum, were assigned to one of four treatments: (i) 3mg norgestomet (N) implant with 5mg estradiol valerate (EDV) and 3mg N injection at the time of insertion (Crestar) for 5 days followed by 600 IU eCG at the time of implant removal; (ii) Crestar for 5 days as in (i) followed by 0.75 mg estradiol benzoate (EDB) 24h later; (iii) Crestar for 9 days followed by 600 IU eCG at the time of implant removal; and (iv) Crestar for 9 days followed by 0.75 mg EDB 24h later. Ovarian scanning was preformed from 4 days before implant insertion until ovulation and 4 days postovulation to detect the CL. Daily blood samples were collected from day 20 post-partum until second ovulation for FSH and E(2) assay. Data were analyzed using analysis of variance. There was no effect of the stage of follicle wave at the time of implant insertion on interval to new follicle wave emergence (range 1-7 days; mean 4.7 days). FSH concentrations were decreased to 5.9+/-2.0 and 7.7+/-1.1 ng/ml for pre- and post-selection cows 1 day after start of treatment; thereafter, they increased on Day 2 to 7.9+/-2.0 and 11.0+/-1.1 ng/ml and on Day 3 to 10.3+/-2.7 and 11.4+/-1.7 ng/ml for pre- and post-selection cows, respectively, despite high-estradiol concentrations at that time. There was no effect of treatment on the interval from implant removal to ovulation (3.2-4.0 days) or on the number of cows detected in estrus (26 of 27 cows). The size of the ovulatory follicle in cows given 0.75 mg EDB 24h post implant removal was decreased in animals at the pre-selection stage (12.2+/-0.1mm) of the follicle wave compared with those at the post-selection stage (15.3+/-0.9 mm) at implant removal. Cows given 600 IU eCG at the pre-selection phase of follicular growth had multiple ovulations (4.0+/-1.1). Cows given EDV at the start of a 5-day implant period had higher estradiol concentrations before and on the day of implant removal than those given EDV at the start of a 9-day implant period. The injection of 0.75 mg EDB 1 day after implant removal tended to increase concentrations of estradiol one day later. In conclusion, 5mg EDV and 3mg N at insertion of a 3mg N implant resulted in variable new follicle wave emergence 1-7 days later in post-partum beef cows nursing calves (22 of 27); both eCG and EDB were equally effective at inducing estrus after implant removal in cows in good BCS, but eCG resulted in a significant increase in ovulation rate in cows treated before dominant follicle selection.

Abstract: The objective of this study was to determine an appropriate exogenous dose of bovine corticotropin-releasing hormone (bCRH) to stimulate the physiological effects of the hypothalamic-pituitary-adrenal axis in steers as a method to test the sensitivity of the pituitary and adrenal gland. Twenty 14-mo-old Holstein-Friesian steers were blocked by weight (443.7+/-2.5 kg) and randomly allotted to receive either saline (control) or bCRH (0.1, 0.3, 1.0, or 1.5 microg/kg BW). Animals were housed in a slatted-floor facility (n = 5 per pen). Indwelling jugular catheters, for both the administration of bCRH and blood collection, were fitted on d -1 of the experiment. Saline and bCRH were administered i.v. at time 0. Serial blood samples were collected at -15, 0, 15, 30, 45, 60, 75, 90, 105, 120, 135, 150, 165, and 180 min relative to time 0. Following administration of 0.1 microg of bCRH/kg BW, the peak ACTH response was not significantly different from pretreatment baseline concentrations (mean concentrations as measured at -15 and 0 min before bCRH administration). Mean ACTH concentrations from 0 to 180 min following 0.1 microg of bCRH/kg BW were not significantly different (P = 0.177) from controls. Administration of 0.3, 1.0, and 1.5 microg of bCRH/kg BW increased (P < 0.05) peak ACTH above pretreatment concentrations, and mean ACTH from 0 to 180 min for these treatments were greater (P < 0.05) than for controls. Peak cortisol responses to all bCRH treatments were greater (P < 0.05) than those to pretreatment concentrations. Mean cortisol concentrations from 0 to 180 min were greater (P < 0.05) in all bCRH-treated steers than in controls, but there were no significant differences among the bCRH treatments. The ratio of mean cortisol to mean ACTH for all bCRH doses tested differed (P < 0.05) from control values, indicating reactivity of the adrenals. In conclusion, bCRH challenge may be a useful method for testing the sensitivity of the hypothalamic-pituitary-adrenal axis in steers subjected to stressful husbandry conditions, and a minimum dose of 0.3 microg of bCRH/kg BW is required to stimulate physiological effects of stressor hormones.

Abstract: To determine the effect of repeated ketoprofen (K) administration to surgically castrated bulls on cortisol, acute-phase proteins, immune function, feed intake, growth and behavior, 50 Holstein x Friesian bulls (11 mo old; 300 +/- 3.3 kg) were assigned to one of five treatments: 1) untreated control (C); 2) surgical castration at 0 min (S); 3) S following an i.v. injection of 3 mg/kg of BW of K at -20 min (SK1); 4) S following 1.5 mg/kg of BW of K at -20 and 0 min (SK2); or 5) S following 1.5 mg/kg of BW of K at -20 and 0 min and 3 mg/kg of BW of K at 24 h (SK3). Castration acutely increased plasma cortisol concentrations in S- and K-treated animals compared with C, with no differences in peak and interval to peak cortisol responses among the castration groups. Overall, the integrated cortisol response was greater (P < 0.05) in the castrates than in C, whereas K treatments decreased (P < 0.05) this response compared with S alone, with no differences between K treatments. Plasma haptoglobin and fibrinogen concentrations were increased (P < 0.05) on d 3 in the castration groups compared with C as the result of tissue trauma induced by castration, whereas SK1 and SK2 had lower (P < 0.05) haptoglobin concentrations than S animals. On d 1, concanavalin A-induced interferon-gamma production was suppressed (P < 0.05) in S and SK3 compared with C, SK1, and SK2 animals. Overall from d 1 to 33, DMI were lower (P < 0.05) in S, SK1, and SK3 than in C animals. From d -1 to 35, ADG were lower (P < 0.05) in S, SK2, and SK3 compared with C animals. A higher (P < 0.05) incidence of standing postures and lower incidence of lying postures was observed in S compared with C during the first 6 h after treatment. However, the higher (P = 0.02) incidence of abnormal standing activities observed for S was reversed (P < 0.05) by the K treatments. In conclusion, surgical castration increased plasma cortisol and acute-phase proteins and decreased immune function, feed intake, and growth rate. Ketoprofen effectively reduced the cortisol response to castration, but there was no advantage in treating with two split doses of K (1.5 mg/kg of BW per dose). A repeated K dose 24 h after treatment (3 mg/kg of BW) had no influence on changes in acute-phase proteins and immune response. Systemic analgesia with K is an effective method for alleviating acute inflammatory stress associated with castration.

Abstract: To determine the effects of burdizzo castration alone or in combination with ketoprofen (K), local anesthesia (LA), or caudal epidural anesthesia (EPI) on plasma cortisol, acute-phase proteins, interferon-gamma production, growth, and behavior of beef cattle, 50 Holstein x Friesian bulls (13 mo old, 307 +/- 5.3 kg) were assigned to (n = 10/treatment): 1) control (handled; C); 2) burdizzo castration (B); 3) B following K (3 mg/ kg of BW i.v.; BK); 4) B following LA (8 mL into each testis and 3 mL s.c. along the line where the jaws of the burdizzo were applied with 2% lidocaine HCl; BLA); and 5) B following EPI (0.05 mg/kg of BW of xylazine HCl and 0.4 mg/kg of BW of lidocaine HCl as caudal epidural; BEPI). The area under the cortisol curve against time was lower (P < 0.05) in BK than in B, BLA, or BEPI animals. On d 1 after treatment, plasma haptoglobin concentrations were higher (P < 0.05) in B, BLA, and BEPI than in BK animals. On d 3, haptoglobin and plasma fibrinogen concentrations were higher (P < 0.05) in all castration groups than in C. On d 7, haptoglobin and fibrinogen concentrations remained higher (P < 0.05) in BLA than in B and C animals. On d 1, concanavalin A-induced interferon-gamma production was lower (P < 0.05) in B, BLA, and BEPI than in C, but there was no difference between BK and C animals. From d -1 to 35, ADG was lower (P < 0.05) in B, BLA, and BEPI animals, but not in BK compared with C animals. Overall, there was a higher (P < 0.05) incidence of combined abnormal postures in B than in C, BK and BEPI animals. Although the use of K and EPI decreased (P < 0.05) these postures compared with B alone or B with LA, there was no difference between the K and EPI treatment. In conclusion, burdizzo castration increased plasma cortisol and acute-phase proteins, and suppressed immune function and growth rates. Local anesthesia prolonged the increase in acute-phase proteins. Ketoprofen was more effective than LA or EPI in decreasing cortisol and partially reversed the reduction in ADG following castration. The use of K or EPI was more effective than LA in decreasing pain-associated behavioral responses observed during the first 6 h after treatment. Systemic analgesia with ketoprofen, a non-steroidal antiinflammatory drug, was more effective in reducing inflammatory responses associated with castration than LA or EPI.

Abstract: Tissue samples were collected postmortem from 126 sheep at five lymphoreticular sites by different techniques. The three most successful combinations of sites and techniques were: the third eyelids, using a forceps and scissors, which provided a mean (se) of 5.32 (0.70) lymphoid follicles per 5 microm tissue section, a mandibular lymph node, using a Biopty gun, which gave 1.19 (0.26) lymphoid follicles per 5 microm tissue section, and tonsil, using a biopsy forceps, which gave 1.14 (0.27) lymphoid follicles per 5 microm tissue section. These three techniques were repeated once a month for five months on five sheep under general anaesthesia, and their clinical effects were compared with five control sheep which were restrained and anaesthetised in the same way but from which no biopsies were taken. Most lymphoid follicles (3.47 [0.58] per 5 pm tissue section) were obtained by using the third eyelid biopsy technique. There were no clinical side effects associated with the biopsy procedure. There were increases in the plasma concentration of cortisol in all the animals, suggesting that the restraint and anaesthesia were more stressful than the biopsy procedure.

Abstract: To determine the effects of the anti-inflammatory ketoprofen, alone or with local anesthesia (LA) during castration on cortisol, immune, and acute phase responses, 40 Friesian calves (215 +/- 3.5 kg) were assigned as follows: 1) control, 2) surgical castration (SURG), 3) SURG following ketoprofen (3 mg/kg BW i.v.; SURG + K), 4) SURG following LA (9 mL of 2% lidocaine hydrochloride to each testis; SURG + LA), or 5) SURG following LA and K (SURG + LA + K). Total cortisol response was greater (P < 0.05) in SURG, SURG + LA, and SURG + K + LA calves than in control calves and was not different between control and SURG + K calves. The interval to peak cortisol was longer (P < 0.05) for SURG + K + LA calves than for either SURG or SURG + K calves. On d 3, KLH-induced interferon-gamma production was lower (P < 0.05) in SURG calves than in control calves, whereas concanavalin A-induced interferon-gamma production was lower (P < 0.05) in all castration groups than in control. On d 1 after surgery, fibrinogen was higher (P < 0.05) in SURG and SURG + LA calves than in control calves, whereas SURG + LA + K calves had lower (P < 0.05) fibrinogen than did SURG calves. Haptoglobin was higher (P < 0.05) in SURG calves on d 1, 3, and 7 than in control calves. On d 1 after surgery, SURG + K and SURG + LA + K calves had lower (P < 0.05) haptoglobin concentrations than SURG calves, whereas SURG + K calves had lower (P < 0.05) levels than SURG calves on d 3. In conclusion, surgical castration induced a significant elevation in cortisol secretion; the rise in cortisol was reduced to control levels by the administration of ketoprofen but not local anaesthetic. Thus, systemic analgesia using ketoprofen is more effective than local anesthesia during castration to alleviate the associated stress response.

Abstract: Ovarian follicle growth in cattle culminates in the selection of a single dominant follicle which attains the ability for final maturation and ovulation once or twice during the luteal phase and at the end of the oestrous cycle, as well as during other reproductive states. This review will describe in detail the first follicle wave of the cycle leading to selection of the first wave dominant follicle, indicating the specific gonadotrophin dependencies of cohort and dominant follicles, and relating follicle fate to steroidogenesis. As a differential gonadotrophin response of growing antral follicles during the follies-stimulating hormone (FSH) decline may determine which follicle becomes selected, first wave follicles are also characterized in relation to intrafollicular growth factors, which may modify the gonadotrophin response, such as inhibins and members of the insulin-like growth factor (IGF) family. Subsequently, the follicular control of the transient FSH rise and decline so crucial to dominant follicle selection will be discussed. It is concluded that successful hormonal manipulation of follicle wave growth and dominant follicle selection will depend on our detailed understanding of the gonadotrophin requirements of differentiating wave follicles.

Abstract: The aim was to determine the effect of estradiol benzoate (EDB) given after removal of a progesterone-releasing intravaginal device (PRID) at either emergence or dominance of a follicle wave, on the interval to estrus, variation in its onset and pregnancy rate in heifers. Heifers (n=186) were assigned randomly to four treatments in a 2 x 2 factorial design; emergence or dominance of a follicle wave at PRID removal, with or without 0.5 mg EDB 24 h after PRID removal. Ovarian ultrasonography was performed to confirm follicular status; data from heifers of undeterminable follicular status were excluded (n=36). Mean size of the largest follicle of the new wave at PRID removal was smaller (P < 0.01) in heifers given EDB at emergence (6.3 +/- 0.09 mm) compared with those given it at dominance (10.9 +/- 0.30 mm). The onset of estrus was earlier (P < 0.01) in heifers given EDB at dominance (median 42 h, range 13 h) compared with those not given EDB at dominance (median 43 h, range 42 h). The median interval to estrus was decreased (P < 0.01) in heifers given EDB at emergence (median 48 h, range 73 h) compared with those not given EDB at emergence (median 66 h, range 45 h). Variation in onset of estrus was reduced (P < 0.05) in heifers given EDB compared with those not given EDB. The pregnancy rate was not affected when EDB was given at dominance, however, it was decreased (P < 0.05) when given at emergence (23 of 40 vs 26 of 32, respectively). To determine the effect of EDB on follicular dynamics in heifers treated with EDB at emergence, heifers (n=37) were assigned to two treatments: at emergence with or without EDB and their ovaries were examined daily using ultrasonography. Follicular dynamics were not different (P > 0.05) in EDB-heifers compared with untreated controls. Mean serum estradiol was greater (P < 0.01) in EDB-treated heifers compared with controls. In conclusion, 0.5 mg EDB given 24 h after PRID removal to heifers decreased the interval to estrous onset at emergence or dominance, decreased variation in onset of estrus and decreased pregnancy rates when given at emergence of a follicle wave.

Abstract: To evaluate the roles of FSH and LH in follicular growth, GnRH-immunized anestrous heifers (n = 17) were randomly assigned (Day 0) to one of three groups (n = 5 or 6). Group 1 received i.m. injections of 1.5 mg porcine FSH (pFSH) 4 times/day for 2 days; group 2 received i.v. injections of 150 microg pLH 6 times/day for 6 days; group 3 received both pFSH and pLH as described for groups 1 and 2. After slaughter on Day 6, measurements were made of follicle number and size, and follicular fluid concentrations of progesterone (P(4)), estradiol (E(2)), and aromatase activity. Injection of pFSH increased (P: < 0.01) the serum concentrations of FSH between 12 and 54 h. Infusion of pLH increased (P: < 0.05) mean and basal concentrations of LH and LH pulse frequency. Serum E(2) concentrations were higher (P: < 0.05) for heifers given pFSH + pLH than those given either pFSH or pLH alone. There was no difference (P: > or = 0.24) between treatments in the number of small follicles (<5 mm). Heifers given pFSH or pFSH + pLH had more (P: < or = 0.02) medium follicles (5.0-9.5 mm) than those that were given pLH alone (none present). Heifers given pFSH + pLH had more (P: = 0.04) large follicles (> or =10 mm) than those given either pLH or pFSH alone (none present). Overall, only 1 of 35 small follicles and 2 of 96 medium follicles were E(2)-active (i.e., E(2):P(4) >1.0), whereas 18 of 21 large follicles (all in the pFSH + pLH treatment) were E(2)-active; of these, 8 of 18 had aromatase activity. Concentrations of E(2) and E(2) activity in follicular fluid were correlated (r > or = 0.57; P: < 0.0001) with aromatase activity in heifers given pLH + pFSH. In conclusion, pLH failed to stimulate follicle growth greater than 5 mm; pFSH stimulated growth of medium follicles that were E(2)-inactive at slaughter and failed to increase serum E(2) concentrations; whereas pFSH + pLH stimulated growth of medium follicles and E(2)-active large follicles, and a 10- to 14-fold increase in serum E(2) concentrations.

Abstract: The aim was to compare the estrous response in heifers given either gonadotropin-releasing hormone (GnRH) or estradiol benzoate (EDB) at the start of a progesterone treatment initiated at emergence or dominance of the first or second follicular wave of the estrous cycle. Cross-bred beef heifers (n=134) were assigned to 1 of 3 treatments; 0.75 mg EDB given at insertion of a progesterone-releasing intravaginal device (PRID) treatment of 10 days duration (10dE2), 0.75 mg EDB at insertion of a PRID treatment of 8 days duration with 15 mg luprostiol (PGF) a luteolytic agent, given 1 day before PRID removal (8dE2) or 250 microg GnRH at insertion of a PRID treatment of 8 days duration with 15 mg PGF given 1 day before PRID removal (8dGnRH). Treatments were initiated on Days 2, 5, 10 or 13 of the estrous cycle. Estrous detection was conducted six times daily. Twice daily blood samples were taken, from 2 days before PRID insertion until detection of estrus. The proportion of heifers detected in estrus was higher (P < 0.05) for heifers in the 8dE2 treatment group (40/40) compared with those in the 8dGnRH group (38/42) and tended to be higher (P = 0.08) than heifers in the 10dE2 group (38/41). The onset of estrus was earlier (P < 0.05) for heifers in the 10dE2 treatment group (median 41 h, range 92 h) compared with either the 8dE2 (median 49 h, range 64 h) or 8dGnRH groups (median 49 h, range 92 h). Submission rate at 72 h was higher (P < 0.01) in the 8dE2 (95%) group than for those in the 10dE2 (74%) and 8dGnRH (69%) groups. In conclusion, EDB given at PRID insertion, with PGF given 1 day before PRID removal, was more effective at synchronizing estrus than was GnRH at PRID insertion. Decreasing the length of treatment and the use of PGF 1 day before the end of an EDB and progesterone treatment improved estrous synchrony.

Abstract: Prolonged postpartum anoestrus in beef cows is due to failure of early dominant follicles to ovulate. It is hypothesized that this failure to ovulate is due to inadequate LH pulse frequency. The objective of this study was to determine whether administration of hourly LH pulses would cause the first dominant follicle to ovulate. In Expt 1, 16 cows received either saline (n = 8) or porcine LH (pLH; 50 micrograms h-1; n = 8) as hourly pulses for 3-5 days from the second day of dominance of the first dominant follicle (day 0). In Expt 2, 21 cows received either saline (n = 7), or 50 micrograms pLH (n = 7) or 100 micrograms pLH (n = 7) as hourly pulses for 3 days. Appropriate ovarian scanning and assays of blood samples were carried out. In Expt 1, the number of dominant follicles that underwent atresia was not affected by increasing the number of LH pulses, but the duration of dominance (days) of the first and second dominant follicles and maximum size (mm) of the second dominant follicle were increased (P < 0.05). Oestradiol concentrations were higher (P < 0.05) in cows given hourly pLH pulses (3.1 +/- 1.2 pg ml-1) compared with controls (1.2 +/- 0.2 pg ml-1). Four of eight treated cows had an anovulatory LH surge. The number of follicle waves to first ovulation was not different (P < 0.05) between control (4.6 +/- 0.9) and pLH treated cows (3.9 +/- 0.5). In Expt 2, four of seven cows given pulses of 100 micrograms pLH h-1 ovulated the first dominant follicle, and the interval from calving to first ovulation was decreased (P < 0.05). In the remaining three cows, the duration of dominance of the first dominant follicle was increased (P < 0.005), the maximum size of the first dominant follicle was greater (P < 0.05), and the interval (days) from the start of infusion to new wave emergence was greater (P < 0.05) compared with cows that failed to ovulate in either the 50 micrograms pLH h-1 or control treatments. In conclusion, hourly pulses of pLH from day 1 after dominance of the first dominant follicle in postpartum beef cows can either prolong dominance or induce it to ovulate. This finding supports the hypothesis that LH pulse frequency is a key determinant of the fate of the dominant follicle in the early postpartum period.

Abstract: To test the hypothesis that emergence of follicle waves postpartum is associated with a change in circulating FSH isoform distribution, 10 Limousin-cross suckler cows were blood sampled daily from 5 wk prepartum until first ovulation postpartum for FSH, LH, estradiol (E2), and progesterone assay. Follicular growth was monitored daily by ultrasonography from Days 5 to 10 postpartum until first ovulation. Distributions of circulating FSH isoforms were characterized (n = 4 per group) by chromatofocusing at 1) 18-33 days prepartum, 2) 3-5 days prepartum, 3) the first postpartum FSH rise responsible for emergence of the first follicle wave, and 4) the FSH rise that stimulated the ovulatory follicle wave. The interval to detection of the first postpartum dominant follicle (DF) was 9.6 +/- 0.58 days. The number of DF before first ovulation was 2.1 +/- 0.18, and first ovulation occurred at 28.6 +/- 1.54 days postpartum. Serum E2 concentrations were higher (p = 0.0001) in cows during the 5-wk period prepartum (53.8 +/- 6.29 pg/ml) than in the postpartum period up to first ovulation (1.5 +/- 0.15 pg/ml). In late pregnancy, there was an absence of recurrent FSH rises and LH concentrations were decreased (p < 0.0001) compared with those in the postpartum period. The emergence of each follicle wave postpartum was preceded by a 2- to 4-day rise in FSH concentrations. The pattern of FSH isoform distribution did not differ (p > or = 0.75) between the pre- and postpartum periods.

Abstract: The objective of this study was to determine the effects of castration, with its presumed pain and inflammatory effects, including increased cortisol, and elevated cortisol per se on in vitro interferon-gamma (IFN-gamma) production, ADG, ADFI, and plasma haptoglobin and fibrinogen. Thirty Friesian bull calves (174 +/- 3.8 kg) were assigned to three treatments (given on d 0): 1) control (CON); 2) i.v. cortisol administration to mimic castration-induced increases in cortisol (CORT); and 3) surgical castration (SURG). Blood samples were collected for 12 h on d 0 and at 24 and 72 h after treatment for cortisol determination. Keyhole limpet hemocyanin (KLH)- and concanavalin A (Con A)-induced in vitro IFN-gamma production in blood, and plasma haptoglobin and fibrinogen were measured in blood samples taken before treatment on d 0 and on d 1 and 3. On d 0, CORT and SURG animals had higher peak cortisol (P < .001) and area under the cortisol curve (P < .001) than CON animals. There were no differences (P > .05) between CON, CORT, and SURG animals in cortisol at 24 and 72 h. There were no differences (P > .05) between CON and CORT animals in IFN-gamma production, haptoglobin, fibrinogen, ADG, and ADFI. Compared with CON animals, SURG animals had lower (P < .05) KLH-induced IFN-gamma on d 1 and CON A-induced IFN-gamma on d 1 and 3. Haptoglobin concentrations were greater (P < .05) for SURG than for CON animals on d 1 and 3. Fibrinogen concentrations were greater (P < .001) for SURG than for CON animals on d 3. The SURG animals had lower (P < .01) ADG and ADFI during d 0 to 7 than CON animals. In conclusion, castration decreased IFN-gamma production, ADG, and ADFI and increased haptoglobin and fibrinogen, and these effects seemed to be independent of plasma cortisol concentrations.

Abstract: The objective was to determine the effects of reducing the plasma cortisol rise in calves following castration on plasma ACTH concentrations, keyhole limpet hemocyanin (KLH)- and concanavalin A (Con A)-induced in vitro interferon (IFN)-gamma production, white blood cell (WBC) numbers, neutrophil:lymphocyte (N:L) ratio, plasma haptoglobin and fibrinogen concentrations, ADG, and ADFI. Forty 5-mo-old Friesian bull calves (169 +/- 1.7 kg) were assigned to four treatments: 1) control (CON); 2) oral metyrapone administration (MET); 3) surgical castration at 0 h on d 0 (SURG); and 4) oral metyrapone administration and surgical castration (MET+SURG). Cortisol, ACTH, IFN-gamma production, haptoglobin, fibrinogen, ADFI, and ADG were not different between CON and MET animals. The MET+SURG calves had lower (P < .001) peak and mean cortisol during .25 to 1.5 h than SURG animals, but area under the cortisol vs time curve from 0 to 12 h did not differ (P > .39) between SURG and MET+SURG calves. Peak ACTH concentrations and area under the ACTH vs time curve from 0 to 6 h were greater (P < .05) for MET+SURG than for SURG calves. There were no differences between MET+SURG and SURG animals in IFN-gamma production, WBC numbers, and ADFI. On d 1, MET+SURG and SURG animals had lower (P < .01) KLH- and Con A-induced IFN-gamma production and higher (P < .05) neutrophil numbers and N:L ratio compared with CON animals. Plasma haptoglobin on d 1 and 3 and fibrinogen concentrations on d 3 and 7 were elevated (P < .05) for MET+SURG and SURG compared with CON animals, whereas SURG animals had greater (P < .05) haptoglobin and fibrinogen concentrations than MET+SURG animals on d 7. The ADG of SURG calves was lower (P < .05) than that of MET+SURG calves during d 0 to 7. Metyrapone treatment partially suppressed cortisol and increased ACTH in castrated calves but did not alter the castration-induced suppression of IFN-gamma and increases in neutrophil numbers and the N:L ratio.

Abstract: Meaningful biological interpretation of the role of follicle-stimulating hormone (FSH) requires use of a validated radioimmunoassay (RIA) that closely estimates biologically active FSH, which was the objective of this work. Three FSH antibodies [NIDDK anti ovine FSH (oFSH); JAD anti oFSH; USDA anti bFSH] were screened against three tracer preparations [USDA oFSH-19-SIAFP-I2(USDA oFSH I2); LER1976a oFSH; USDA bFSH I2] in a RIA using USDA bFSH B1 or I2 as the assay standard. Sera obtained from three heifers at 4- to 8-h intervals for 5 days after injection of PGF2 alpha during the luteal phase were assayed for both FSH immunoactivity using each of the three optimized assay formats (NIDDK anti oFSH and JAD anti oFSH with USDA oFSH I2 as tracer; USDA anti bFSH with USDA bFSH I2 as tracer), and FSH bioactivity, using a rat Sertoli cell bioassay. Cross reactivity of bLH (NIH bLH B9) in all three assay formats was minimal (0.7, 0.9 and < 0.4% at 50% binding for the NIDDK, JAD and USDA antibodies, respectively). There was parallel displacement of tracer between bovine serum dilutions of 10 to 500 microliters and the two FSH standards. Correlations between JAD and USDA RIA data and bioassay results were not significant (P > or = 0.10), but were significant (r = 0.78; P = 0.0001) for the NIDDK RIA FSH and the bioactive FSH measurements. The assay sensitivity of the NIDDK RIA was 0.55 ng USDA bFSH B1 (0.013 ng USDA bFSH I2)/ml. The inter- and intra-assay CV were between 5.8 and 7.9 %. This RIA detected a pre-ovulatory FSH surge coincident with the LH surge, in all heifers studied. Furthermore, the emergence of each wave of follicle growth (up to day 12 of the cycle), was preceded by a transient increase (P < 0.02; days 0.5 to 1.5 and 8 to 10.5 of the cycle) in serum FSH, while LH concentrations remained unchanged. In conclusion, the RIA utilising NIDDK anti oFSH and USDA oFSH I2 as tracer provides a good estimate of bioactive FSH in cattle, and detects physiologically relevant increases in serum FSH related to emergency of each new wave of follicle growth.

Abstract: Blood samples were collected from heifers (n = 6; 450 +/- 7.7 kg) 2-4 times a day during the first and second follicular waves, and during the gonadotrophin surge to determine whether changes in circulating FSH isoforms occur during cyclic quantitative changes in FSH throughout the oestrous cycle. Serum was assayed for FSH, LH, oestradiol and progesterone. Selected samples collected during the first (samples 1-3) and second (samples 4-6) postovulatory recurrent FSH increase and during the subsequent gonadotrophin surge (samples 7 and 8) were analysed for FSH isoforms by chromatofocusing. No change (P > 0.05) in isoform profile occurred during the first or second recurrent FSH increase, when oestradiol concentrations were 0.6 +/- 0.07 and 0.6 +/- 0.09 pg ml-1 and progesterone concentrations were 0.03 +/- 0.01 and 2.4 +/- 0.19 ng ml-1, respectively. The percentage of FSH eluting in the pH range 7.4-7.0 increased (P < 0.05) from 14.2 +/- 2.2 during the luteal phase (samples 1-6) to 20.2 +/- 2.3 (sample 7) and 31.4 +/- 3.4% (sample 8) during the preovulatory gonadotrophin surge, while oestradiol concentrations were higher (P < 0.05; 4.9 +/- 0.39 pg ml-1) than in the luteal phase of the cycle. In summary, FSH isoform patterns did not change during the cyclic quantitative changes in FSH associated with emergence of the first or second follicular wave. However, during the gonadotrophin surge, in association with increased oestradiol concentrations, an increase in the amount of less acidic isoforms of FSH was observed. Therefore, qualitative changes in FSH are not important in the physiological regulation of follicle turnover during the luteal phase of the oestrous cycle of heifers.

Abstract: To determine the effects of castration of calves, with or without local anesthesia, on plasma cortisol, scrotal circumference, ADG, and ADFI, 56 Friesian bulls (5.5 mo of age; mean +/- SE BW = 173 +/- 2 kg) were randomly assigned to each of seven treatments: 1) control (CON); 2) s.c. injection of .1 mg of a human serum albumin-GnRH conjugate with DEAE-dextran adjuvant (HSA-GnRH); 3) burdizzo castration without local anesthetic (BURD); 4) burdizzo castration following local anesthetic administration (BURD + LA); 5) surgical castration without local anesthetic (SURG); 6) surgical castration following local anesthetic administration (SURG + LA); and 7) local anesthetic administration alone (LAA). Blood samples for cortisol analyses were taken via jugular catheter from -2 to 10 h and at 24, 48, and 72 h relative to treatment. Average daily feed intakes were recorded for 5-d periods and calves weighed at 7-d intervals before and after treatment. Local anesthetic alone had no effect (P > .10) on any variable. The HSA-GnRH calves had elevated (P < .05) plasma cortisol from 2 to 6 h compared with CON calves. Peak plasma cortisol was elevated (P < .01) in BURD, BURD + LA, SURG, and SURG + LA compared with CON calves. The SURG calves (46.0 ng/mL) had higher (P < .03) peak cortisol than BURD (31.4 ng/mL) and SURG + LA (35.4 ng/mL) calves. There was no difference in peak cortisol between BURD and BURD + LA (26.5 ng/mL) calves. The ADG from d 0 to 7 was reduced (P < .05) in calves in BURD + LA, SURG, and SURG + LA treatments (-.01, -.83 and -.24 kg, respectively) compared with CON calves (.54 kg). The ADFI were reduced (P < .05) in BURD and BURD + LA calves during d 1 to 5 and in BURD + LA, SURG, and SURG + LA calves during d 6 to 10 compared with CON calves. The scrotal circumferences of BURD and BURD + LA calves were greater (P < .05) than those of CON calves for 7- and 35-d periods post-castration, respectively. Castration induced increases in cortisol and decreases in ADG and ADFI. Surgical castration induced a greater plasma cortisol response than burdizzo castration, and the administration of local anesthetic reduced the cortisol response of surgical castrates but was less effective for burdizzo castrates.

Abstract: The objective was to determine the effect of gonadotrophin-releasing hormone (GnRH), GnRH analogue (GnRH-A) or oestradiol administration on luteinising hormone (LH) and follicle-stimulating hormone (FSH) release in GnRH-immunised anoestrous and control cyclic heifers. Thirty-two heifers (477 +/- 7.1 kg) were immunised against either human serum albumin (HSA; controls; n = 8), or a HSA-GnRH conjugate. On day 70 after primary immunisation, control heifers (n = 4 per treatment; day 3 of cycle) received either (a) 2.5 micrograms GnRH or (b) 2.5 micrograms of GnRH-A (Buserelin) and GnRH-immunised heifers (blocked by GnRH antibody titre; n = 6 per treatment) received either (c) saline, (d) 2.5 micrograms GnRH, (e) 25 micrograms GnRH or (f) 2.5 micrograms GnRH-A, intravenously. On day 105, 1 mg oestradiol was injected (intramuscularly) into control (n = 6) and GnRH-immunised anoestrous heifers with either low (13.4 +/- 1.9% binding at 1:640; n = 6) or high GnRH antibody titres (33.4 +/- 4.8% binding; n = 6). Data were analysed by ANOVA. Mean plasma LH and FSH concentrations on day 69 were higher (P < 0.05) in control than in GnRH-immunised heifers (3.1 +/- 0.16 vs. 2.5 +/- 0.12 ng LH ml-1 and 22.5 +/- 0.73 vs. 17.1 +/- 0.64 ng FSH ml-1, respectively). The number of LH pulses was higher (P < 0.05) in control than in GnRH-immunised heifers on day 69 (3.4 +/- 0.45 and 1.0 +/- 0.26 pulses per 6 h, respectively). On day 70, 2.5 micrograms GnRH increased (P < 0.05) LH concentrations in control but not in GnRH-immunised heifers, while both 25 micrograms GnRH and 2.5 micrograms GnRH-A increased (P < 0.05) LH concentrations in GnRH-immunised heifers, and 2.5 micrograms GnRH-A increased LH in controls. FSH was increased (P < 0.05) in GnRH-immunised heifers following 25 micrograms GnRH and 2.5 micrograms GnRH-A. Oestradiol challenge increased (P < 0.05) LH concentrations during the 13-24 h period after challenge with a greater (P < 0.05) increase in control than in GnRH-immunised heifers. FSH concentrations were decreased (P < 0.05) for at least 30 h after oestradiol challenge. In conclusion, GnRH immunisation decreased LH pulsatility and mean LH and FSH concentrations. GnRH antibodies neutralised low doses of GnRH (2.5 micrograms), but not high doses of GnRH (25 micrograms) and GnRH-A (2.5 micrograms). GnRH immunisation decreased the rise in LH concentrations following oestradiol challenge.

Abstract: To investigate the effects of active immunization of cyclic beef heifers with different doses of a human serum albumin-Cys-Gly-GnRH (HSA-GnRH) conjugate on antibody titers, ovarian function, body growth, and carcass characteristics, 32 heifers (BW = 477 +/- 7.1 kg; mean +/- SE) were assigned to one of four immunization treatments: .1 mg of HSA or .01, .1, or 1.0 mg of HSA-GnRH, respectively. All heifers received a primary (d 0) and booster (d 28) immunization using DEAE-dextran as adjuvant. The duration of the experiment was 158 d. Overall antibody titers against GnRH were greater (P < .05) for heifers immunized against GnRH (13 +/- 3.3, 22 +/- 3.8, and 19 +/- 2.8% binding at a plasma dilution of 1: 640 for Treatments 2 to 4, respectively) than for controls (1 +/- .1%). The numbers of heifers that became anestrous (plasma progesterone < .5 ng/mL for > 21 d) were 1/8, 8/8, 7/8, and 8/8, respectively. The interval from primary immunization to anestrus (40.7 +/- 6 d) and the duration of anestrus (78 +/- 7 d) were not affected by dose of HSA-GnRH conjugate. The number of ovulations detected was reduced (P < .05) in GnRH-immunized (4.6 +/- .64, 4.0 +/- .70, and 3.6 +/- .60 for Treatments 2 to 4, respectively) compared with control heifers (9.4 +/- .20). During induced anestrus, follicular growth was generally arrested (< 5 mm in diameter) and plasma estradiol decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: To determine the effects of active immunization against prostaglandin F2 alpha (PGF) on estrous activity and performance traits of beef heifers, 50 14-mo-old cyclic heifers (358 +/- 3.3 kg) were assigned to two treatments (n = 25 per treatment): 1) heifers (controls) given 3.3 mg of human serum albumin (HSA) on d 0 (primary) and 27 (booster), and 2) heifers (PGF-immunized) given 3.3 mg of PGF-HSA on d 0 and 27. The adjuvant was DEAE-dextran, and the duration of the experiment was 167 d. Plasma progesterone concentrations (every 3 to 4 d) were used to monitor corpus luteum (CL) presence; PGF antibody titers were determined every 2 wk. Heifers were checked twice daily for estrous behavior and were weighed every 2 wk. Data were analyzed using ANOVA. Antibody titers for PGF-immunized heifers increased to a peak (43 +/- 2.9% binding at a plasma dilution of 1:1,250) on d 55 +/- 4.6. Antibody titers were greater (P = .02) in PGF-immunized than in control heifers by d 15 and remained elevated (P < or = .001) throughout the experiment. Twenty-four of 25 PGF-immunized heifers formed persistent CL with a mean duration of 129 +/- 6.4 d. The mean number of estrous period per heifer were less for PGF-immunized (1.5 +/- .27) than for control heifers (7.0 +/- .32). Mean daily live weight gain of the PGF-immunized heifers was decreased (P < .05; .75 +/- .024 kg) compared with that of controls (.83 +/- .014 kg), largely due to a 31.5% decrease during the 28-d period after booster.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Growth rate of heifers is reduced by prostaglandin F2 alpha (PGF) immunization following a primary and booster regimen. The objective was to attenuate the immune response with or without a booster immunization; specifically, the effects of booster interval, dose of PGF-human serum albumin (HSA) conjugate at booster, adjuvant type, or single immunization with one or two adjuvants were examined. Three experiments were conducted using 175 cyclic heifers. Plasma PGF antibody titers were measured every 2 wk and progesterone concentrations every 3 to 4 d. In Exp. 1, single immunization with one adjuvant (3.3 mg of PGF-HSA in either DEAE-dextran [DEAE] or non-ulcerative Freund's adjuvant [NUFA]; or 10.0 mg of PGF-HSA in NUFA) did not induce sufficient antibody titers to consistently induce persistent corpora lutea (CL). Booster intervals of either 14, 21, or 28 d increased titers sufficiently to induce persistent CL (34/35 heifers), but ADG of heifers was less (P < .05) than for those given a single immunization. In Exp. 2, 1.0 mg of conjugate for booster immunization induced a greater (P < .05) immune response than 3.3 mg, and both doses decreased (P < .05) ADG. Single immunization, with half the conjugate dose in DEAE and half in NUFA injected separately, induced persistent CL in 7/8 heifers without decreasing ADG compared with controls. In Exp. 3, single immunization, with half the conjugate dose in DEAE and half in NUFA injected separately, prolonged (P < .05) the intervals to peak titer compared with the booster treatment, but the incidence (13/15 vs 8/8) and duration (120 +/- 4.8 vs 111 +/- 7.9 d) of persistent CL were similar, and ADG was greater (P < .05). In conclusion, attempts to attenuate the immune response following booster immunization were unsuccessful. Single immunization, using two adjuvants separately, induced persistent CL for at least 120 d without decreasing ADG compared with the primary and booster regimen.

Abstract: To develop an effective immunization protocol against human serum albumin-Cys-Gly-GnRH (HSA-GnRH) conjugate to delay the onset of puberty in heifers, 58 heifers (8 mo of age; mean +/- SE BW = 203 +/- 1 kg) were randomly assigned to each of six treatments: 1) controls, .1 mg of HSA, with diethylaminoethyl (DEAE)-dextran as adjuvant, on d 0 and 28; 2) .1 mg of HSA-GnRH, with DEAE-dextran, on d 0; 3) as 2) and booster on d 28; 4) as 3) but boosters also on d 84, 140, 196, and 252; 5) as 2) but half the conjugate given with DEAE-dextran adjuvant and half with non-ulcerative Freund's adjuvant (NUFA), injected in two separate sites; and 6) as 2) but the conjugate given with DEAE-dextran and NUFA, emulsified and injected in two sites. The duration of the experiment was 342 d. Mean plasma GnRH antibody titers (samples every 2 wk) for heifers in Treatments 2 to 6 were 9.4 +/- 1.16, 20.6 +/- 2.21, 43.9 +/- 2.86, 27.9 +/- 2.67, and 44.5 +/- 3.75% binding at a plasma dilution of 1:640. The mean number of times estrus was observed in heifers was less (P < .05; pooled SEM = .53) in Treatments 4 (.2) and 6 (2.4) than in Treatments 1, 2, 3, and 5 (7.8, 7.0, 7.0, and 6.6, respectively). The mean interval to the onset of puberty (the first increase in plasma progesterone > or = .5 ng/mL for > or = 10 d with samples at 3- to 4-d intervals) was greater (P < .05; pooled SEM = 11.6) for heifers in treatments 4 (339 d) and 6 (276 d) than for heifers in Treatments 1, 2, 3, and 5 (164, 159, 165, and 170 d, respectively). Mean ADG of heifers was reduced (P < .05) in treatments 2, 3, 4, and 6 (.71, .72, .68, and .69 kg, respectively) compared with controls (.77). In summary, the multiple booster immunization treatment induced and maintained sufficient anti-GnRH titer to delay puberty for 175 d; a single immunization against GnRH with DEAE and NUFA increased antibody titers enough to delay puberty for 112 d. However, GnRH immunization treatments reduced ADG of heifers in Treatments 2, 3, 4, and 6.

Abstract: To optimize the prostaglandin F2 alpha (PGF) immunization protocol (conjugate [PGF-human serum albumin; PGF-HSA] dose and immunization regimen) to achieve prolonged suppression of estrous behavior (EB) in beef heifers, 56, 14-mo-old cyclic heifers were assigned (n = 7 per treatment) to eight treatments: 1) 3.3 mg of PGF-HSA on d 0 (single); 2) 3.3 mg of PGF-HSA on d 0 and 28 (booster; B); 3) as (2) except on d 0 and 55; 4) as (2) except on d 0 and 83; and 5 to 8) as in Treatments 1 to 4 except using 10 mg of PGF-HSA. The adjuvant was diethylaminoethyl-dextran, and duration of the experiment was 170 d. Heifers were checked twice daily for EB. A persistent corpus luteum (CL) was considered present when progesterone (P4) was > or = .5 ng/mL for > or = six consecutive samples (every 3 to 4 d). Data were analyzed using ANOVA for a factorial plan. All heifers produced plasma antibody titers (samples every 2 wk) against PGF (peak range: 7 to 84% binding at 1: 1,250). There were no effects (P > .10) of conjugate dose and no interactions between dose and immunization regimen for any variable; therefore, data were combined across dose. Mean and peak titers were greater (P < .05) in heifers in 55- and 83-d B treatments than those in single immunization and 28-d B treatments. Overall, 48/55 heifers formed a persistent CL (41/41 for B heifers). In the single, and 55- and 83-d B treatments, 23/42 heifers formed persistent CL in response to single/primary immunization. There was no difference between immunization regimens in duration (133 +/- 4.4 d) of persistent CL.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: This study examined the correlation between measurement of follicle growth by ultrasound, and measurement of intrafollicular ratios of oestradiol and progesterone concentrations and the serum concentrations of FSH during selection, dominance and atresia or ovulation of dominant follicles in heifers. Heifers were ovariectomized on days 0 (before LH surge), 1 (after LH surge, preovulation), 1 (postovulation), 3, 6 and 12 of the oestrous cycle. Blood samples were collected at 4-6 h intervals. After ovariectomy all follicles > or = 5 mm were measured and follicular fluid was aspirated. Follicles were classified by size according to ultrasound (F1, largest; F2, second largest; F3, all remaining follicles > or = 5 mm) and by the ratio of oestradiol:progesterone concentrations. During the follicular phase, a single dominant oestrogen-active follicle increased in diameter while serum concentrations of LH increased and FSH decreased (P < 0.05). On day 1 (after LH surge, preovulation), serum LH and FSH decreased to pre-surge concentrations (P < 0.0001), while follicle size and intrafollicular progesterone concentration increased and oestradiol concentration decreased (P < 0.05). A dominant nonovulatory follicle, classified as oestrogen-active on days 1, 3 and 6 and oestrogen-inactive on day 12, increased in size from day 1 to day 7 and lost dominance during days 10-12, coincident with the growth of multiple oestrogen-active follicles. The serum FSH concentration increased transiently (P < 0.05) before each new wave of dominant follicular growth. The overall correlation of ultrasound measurements of follicle diameter with measures of follicle size after ovariectomy was high.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Fifty-six prepubertal Hereford cross Friesian heifers were assigned to seven treatment groups: (1) primary (day 0) and booster (day 41) using 10 mg human serum albumin (HSA) (control); (2) primary and booster (day 41) immunizations using 3.3 mg prostaglandin F2 alpha PGF-HSA conjugate; (3) primary and booster (day 83) using 3.3 mg PGF-HSA; (4) primary and booster (day 210) using 3.3 mg PGF-HSA; (5-7) as in treatments 2-4 except 10 mg PGF-HSA were used. Plasma progesterone concentrations were used to determine the onset of puberty and the presence of a corpus luteum (CL); plasma PGF antibody titres were determined at 28-day intervals. There was no effect (P > 0.05) of conjugate dose or booster interval on mean antibody titres and there was no interaction between them. However, heifers in the 83- and 210-day booster treatments had higher (P < 0.05) peak antibody titres than heifers in the 42-day booster treatments. Puberty was delayed in 40% (16/40) of PGF-immunized heifers and 40% (16/40) of heifers formed persistent CL after puberty. Overall, eight of the heifers with delayed puberty also formed a persistent CL. There was a positive correlation between mean titre and onset of puberty but not with duration of persistent CL.

Abstract: There is a low incidence of ovulation of the first dominant follicle that develops in the early postpartum period of beef suckler cows, which prolongs the interval from calving to first ovulation. The objective of this study was to determine whether a single injection of a GnRH analogue would ovulate the first postpartum dominant follicle. Limousin x Friesian beef suckler cows were assigned at parturition, over two years (16 cows in year 1; 19 cows in year 2), to one of three treatments: (1) untreated (control; n = 12), (2) GnRH analogue (20 micrograms buserelin i.m.) administered in the growing-plateau phase of the first postpartum dominant follicle (GnRH-G; n = 12) and (3) GnRH analogue administered in the declining phase of the first postpartum dominant follicle (GnRH-D; n = 11). From day 8 or 9 post partum, the ovaries of each cow were examined daily by ultrasound to determine the time of GnRH injection and ovulation. Blood samples were collected daily for progesterone measurement to confirm ovulation and in year 2 to determine the duration of the first oestrous cycle. The mean (+/- SEM) number of days from parturition to development of the first dominant follicle was 11.0 +/- 0.3, 10.3 +/- 0.5 and 10.1 +/- 0.7 for cows assigned to treatments 1-3, respectively (P > 0.05). The proportion of cows ovulating the first dominant follicle was higher (P < 0.05) following GnRH treatment (12 of 12 and 7 of 10; GnRH-G and GnRH-D, respectively) than with controls (2 of 12).(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: To determine whether systemic and/or intraovarian concentrations of insulin-like growth factor-I (IGF-I) are affected by short-term fasting, 24 heifers were blocked by weight and, within block, were assigned to one of three treatments: fasted for 0 h (controls; n = 8), fasted for 24 h (n = 8), or fasted for 48 h (n = 8). Blood plasma was collected every 8 h from -64 h to 0 h before ovariectomy (OVEX). OVEX was performed per vagina under local anesthesia during the follicular phase of an estrous cycle (36-42 h after synchronization with prostaglandin-F2 alpha). Follicular fluid (FFL) and granulosa cells were collected individually from follicles greater than or equal to 6 mm (large), and FFL was pooled from follicles 1.0-5.9 mm (small) in diameter. Fasting did not affect (p greater than 0.20) the number (mean +/- SE) of small (52 +/- 7) or large (1.5 +/- 0.4) follicles per heifer, specific binding of 125I-hCG to granulosa cells of follicles greater than or equal to 8 mm in diameter, or concentrations of progesterone in FFL of small follicles. At OVEX, body weight was less (p less than 0.01) for 24 h- and 48 h-fasted heifers (412 +/- 7 kg and 399 +/- 7 kg, respectively) than for 0 h-fasted heifers (442 +/- 7 kg). At OVEX, plasma concentrations of IGF-I were lower (p less than 0.05) in the 48 h-fasted group (105 +/- 8 ng/ml) than in the 0 h-fasted group (140 +/- 8 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: Friesian x Hereford heifers (n = 19; mean +/- s.e.m. body weight (BW) = 375 +/- 5 kg) were used in a randomized incomplete block design. Heifers were fed 0.7 (n = 7; L), 1.1 (n = 7; M) or 1.8% (n = 5; G) of BW in dry matter (DM)/day for 10 weeks. Ovaries were examined by ultrasound, for one oestrous cycle, from week 5 of treatment. Maximum diameter of dominant follicles was smaller (P less than 0.05) in L (11.8 +/- 0.1 mm) than in M (13.7 +/- 0.2 mm) or G (13.2 +/- 0.3 mm) heifers. Growth rate (mm/day) of dominant follicles during the oestrous cycle was not affected (P greater than 0.05) by dietary intake. Persistence of dominant follicles was shorter (P less than 0.05) in L (9.8 +/- 0.2 days) than in M (11.9 +/- 0.3 days) or G (12.7 +/- 0.4 days) heifers. Three dominant follicles were identified during the oestrous cycle of 5 of 7 L, 3 of 7 M and 1 of 5 G heifers (P less than 0.10); 2 dominant follicles were identified in the remaining heifers (n = 2 of 7, 4 of 7 and 4 of 5, respectively). Length of the luteal phase and luteal-phase concentrations of progesterone were not affected (P greater than 0.05) by treatment. Low dietary intake reduced the diameter and persistence of dominant follicles during the oestrous cycle of beef heifers and tended to increase the proportion of oestrous cycles with 3 dominant follicles.