Estradiol regulation of luteinizing hormone secretion in heifers of two breed types that reach puberty at different ages

The working hypothesis was that 17 beta-estradiol (E(2)) negative feedback on the hypothalamic-pituitary axis in regulation of LH secretion decreases during peripuberty in heifers of 2 different genotypes. We investigated whether Bos indicus heifers had a period postpuberty, as compared with prepuberty, of greater E(2) inhibition of LH secretion at a time when heifers of this genotype have been reported to have a period of anestrus. Prepubertal heifers 9 mo of age of 2 genotypes (B. indicus and B. taurus) were assigned to 3 groups (6 animals/group) to either remain intact (control), be ovariectomized, or be ovariectomized and implanted with E(2). Variables evaluated from 10 to 28 mo of age were circulating concentrations of progesterone (P(4)), presence of corpora lutea, and pulsatile pattern of LH release. Results confirmed that B. taurus heifers attained puberty at younger ages (P < 0.001) and at lower live weights (P = 0.015) than did B. indicus heifers (507 +/- 37 days of age vs. 678 +/- 7 days of age; 259 +/- 14 kg vs. 312 +/- 11 kg; respectively). There was cessation of E(2) inhibition of LH pulses coincident with the onset of puberty in heifers of both breed types but at a much younger age in B. taurus heifers. There was no evidence of enhanced negative feedback of E(2) on LH secretion subsequent to puberty in B. indicus heifers nor was there cessation of estrous cycles in control heifers of either breed type after puberty.     

Body composition of beef heifers at puberty

Three trials were conducted to study age, weight, percent body fat and protein content of 131 crossbred and purebred beef heifers at puberty, to investigate relationships among various pubertal measurements and to determine if body fat and protein values could be used in combination with age, weight and shoulder height to predict the onset of puberty. Trials 1 and 2 compared heifers on treatments of high (H), medium (M) and low (L) levels of energy. The rations were formulated at 120, 100 and 80 percent of the National Research Council (NRC) recommendation for energy. The protein level was 100 percent of the NRC recommendation for all three treatments. Each animal was examined every two weeks per rectum for the presence of follicles and corpora lutea. The presence of a mature corpus luteum indicated the attainment of puberty. Trial 3 heifers were fed the same level of nutrition but ovulation data were based on weekly rectal palpations and on the presence of greater than 2.0 ng/ml blood plasma progesterone. Percentage of fat and protein were quantified in all trials using a whole body counter. In Trial 1 there was a significant difference (P<0.01) among energy levels for mean weight and percent fat values at puberty, but no differences in age at puberty. Even though heifers in Trial 2 were also fed different energy levels, there were no differences among treatments for the variables measured. The onset of puberty in Trial 2 appeared to be delayed due to cooler than normal weather during that experimental period. Heifers in Trial 2 tended to be older and reached puberty approximately one month later in the year than heifers in Trials 1 and 3, despite similarities in weight gain among the trials. In all trials, high R(2) values for multiple stepwise regression analyses indicated that body composition estimates were useful in predicting weight at puberty in beef heifers. The results of this study do not support a critical body weight or body composition hypothesis in the beef heifer. It was concluded that these data indicate environmental factors may have more effect than nutrition on the onset of puberty.     

Effects of suckling frequency and breed on productive performance,follicular dynamics and postpartum interval in beef cows

One experiment was conducted to determine the influence of suckling frequency (restricted suckling (RS) versus ad libitum suckling (AS)) and breed (Brown Swiss versus Pirenaica) on productive performance, follicular dynamics and postpartum cyclicity resumption in beef cows. Twenty-four spring-calving cows (12 Brown Swiss and 12 Pirenaica) were used. After calving cows were assigned, within breed, to the two suckling frequencies (RS versus AS). During the postpartum period, cows were fed 100% of the daily energy estimated requirements. The study was conducted from calving to either day 90 postpartum or to detection of the second ovulation whenever it came later. The pattern of follicular development was monitored daily by transrectal ultrasonography, and blood samples were collected three times a week to analyse progesterone concentrations by RIA. Body condition score at calving was 2.5 (from 1 to 5). Productive parameters were not significantly affected by suckling frequency. Pirenaica cows gained more weight (P<0.001), while Brown Swiss cows had higher milk yield (P<0.001) and thus their calves grew faster (P<0.05). In the present moderate conditions of nutrition, either suckling frequency or breed did not affect follicular dynamics (on average there were 6.5 follicles in emergence, growth rate of dominant follicle (DF) was 1.3mm per day and maximum diameter of dominant follicle was 13.9mm). There were gradual increases in maximum diameter and growth rate of dominant follicle related to the proximity of the ovulatory waves, and they were defined by third order polynomial equations. Postpartum anoestrous interval (PPI) was markedly affected by the interaction between suckling frequency and breed (P<0.001): ad libitum suckling prolonged significantly the interval from calving to first ovulation in Brown Swiss cows (31.8 day versus 82.7 day, in RS and AS groups, respectively, P<0.05), but not in Pirenaica cows (43.5 day versus 32.6 day, in RS and AS groups, respectively, not statistically significant). It is concluded that in moderate pre- and postpartum nutritional conditions Brown Swiss cows were more susceptible to the suppressor effect of ad libitum suckling on postpartum anoestrous interval than Pirenaica cows, although this breed difference did not seem to be mediated by differences in energy balance or in ovarian follicular dynamics.     

Effect of clitoral stimulation after artificial insemination on conception in Zebu-crossbred heifers in the tropics

In order to increase conception rates to AI, an experiment was conducted to determine the effect of clitoral stimulation on pregnancy rate. Seven hundred and thrity-five Zebu-cross heifers were assigned to either of the following treatments: 1) control without clitoral stimulation (n=351) or 2) clitoral stimulation after IA (n=384). The data were analyzed using a linear model procedure. Clitoral stimulation increased pregnancy rates (57 vs 45 % for treated and control groups, respectively; P<0.02). Holstein x Zebu and Brown Swiss x Zebu heifers attained similar rates (P>0.05). There was a significant treatment-by-season interaction (P<0.01), with pregnancy rates being higher using clitoral massage during the rainy season (54.7 and 25.5% for the treated and control group respectively). In this study, manual clitoral stimulation increased conception rates in Zebu cross heifers under tropical conditions.     

Effects of season of birth on the prepubertal pattern of gonadotropin secretion and age at puberty in beef heifers

There is an early transient rise in gonadotropin secretion in spring-born prepubertal heifers and there is an indication that this pattern is different in autumn-born heifers. The effect of season of birth on age and weight at puberty is equivocal. This study was designed to compare the temporal patterns of LH and FSH secretion between spring- and autumn-born heifers and to determine the effects of season of birth on age and weight at puberty. Blood samples from 2 groups of heifer calves born in spring (last week of March, n = 5) or autumn (last week of October, n = 5) were collected every other week from birth to puberty and every 15 min for 10 h at 6, 12, 18, 24 and 32 wk of age. Timing of puberty was determined by measuring progesterone in plasma samples collected every 2 to 3 d starting at 42 wk of age. Age and weight at onset of puberty did not differ between the 2 groups of heifers (P > 0.05); however, the autumn-born heifers tended to mature in a wider range of ages and weights. Based on the 10-h sampling periods, mean serum concentrations of LH and LH pulse frequency and amplitude were higher in spring-born heifers at 18 wk of age than in autumn-born heifers (P < 0.05). In spring-born heifers, LH pulse frequency increased over time to 32 wk of age, and LH pulse amplitude was higher at 12 and 18 wk than at 32 wk of age (P < 0.05). Autumn-born heifers had higher LH pulse frequency at 6 wk and showed a decrease in mean concentrations of LH at 12 and 18 wk of age (P < 0.05). The FSH pulse frequency of spring-born heifers was higher at 12 wk of age than in autumn-born heifers (P < 0.05), FSH pulse amplitude in autumn-born heifers decreased from 6 to 32 wk of age. It was concluded that although the mean age and weight at puberty did not differ between spring- and autumn-born heifers, the range in age and weight at puberty was wider in the autumn-born heifers. The patterns of LH secretion differed between spring- and autumn-born prepubertal heifers, with spring-born calves exhibiting an early rise in LH secretion, while mean serum concentrations of LH decreased during this period in autumn-born heifers.     

Effect of the collection method on semen characteristics of Zebu and European type cattle in the tropics

The effect of the collection method on the characteristics of fresh semen and the recovery of spermatozoa after thawing was studied in 30 Zebu bulls (Bos indicus ) and in 30 Brown Swiss (Bos taurus ) bulls. Semen was collected by using an artificial vagina and by electroejaculation; the ejaculates were individually evaluated. Semen was diluted for freezing in skimmed milk and stored in 0.5-ml French straws, at a concentration of 30 x 10(6) spermatozoa. Data were evaluated by analysis of variance using a factorial model which included collection method, breed effect, and interaction between method and breed, with each bull as a block. Higher volume and pH of the semen was obtained following electroejaculation. Conversely, higher concentration prior to freezing and better progressive motility after thawing was observed in semen collected with an artificial vagina. No differences in motility were obtained in fresh semen between methods. Better post-thaw recovery occurred when the semen was collected by an artificial vagina, independently of the breed type.     

Serum thyroid hormone concentrations during growth and puberty in Carora dairy heifers of Venezuela

The aim of this study was to characterize changes in serum concentrations of triiodothyronine (T(3)) and thyroxine (T(4)) in relation to growth and the onset of puberty in Carora heifers, a Venezuelan dairy breed. Heifers aged 7 to 16 months were grouped retrospectively according to puberty status into 4 groups: Pubertal (control group; n = 12) and nonpubertal (n = 8) contemporary (born during the same week) heifers, pubertal (n = 7.) and nonpubertal (n = 7) noncontemporary (born in different months) heifers. A split-plot model with repeated measures over Months 7 to 16 of age was used. Control heifers attained puberty at 290 +/- 5 kg body weight (BW) between 13 and 14 months of age. Significant (P < 0.01) interactions of birth time of year and puberty status were detected in BW and serum concentrations of T(3), but only interaction of birth time of year was found in T(4). A transient but significant (P < 0.05) decrease in T(3) secretion was seen in pubertal contemporary and noncontemporary (101 +/- 4 and 113 +/- 4 ng/d1, respectively) heifers at Month 12 of age, a change which could be critical to the onset of puberty. Significant (P < 0.05) positive correlation was found between BW and T(3). In summary, thyroid hormone secretion changed across the months of growing and around the onset of puberty in Carora heifers.     

Reproductive responses of beef heifers and cows to exogenous progesterone administered in silastic coils,oestradiol benzoate and pregnant mare serum gonadotropin

The aim of this study was to investigate a Progesterone Releasing Intravaginal Device, Oestradiol benzoate and PMSG combined treatment in 81 suckling charolais cows and 56 heifers of various breeds. The former were young primiparous cows aged 2 years and treated at a means of 43 days postpartum. Most of the heifers ($\text{44}\text{56}$) were not cycling and were believed to be prepuberal.In non-cycling females, the treatment induced ovulation in 98.5% of the cows and 78 to 100% of the heifers according to breed. In those females in which ovulation was induced, pregnancy rates determined by rectal palpation were 46.9% for the cows and 66.7% for the heifers. When taking into account all treated females including those with lost Progesterone Releasing Intravaginal Devices (5.8%) and those not ovulating, the pregnancy rates determined by rectal palpation 3 to 5 months after artificial insemination were 40.7% for the cows and 52% for the heifers.     

Sexual development of dairy bulls in the Mexican tropics

Sexual development and pubertal traits were studied in Holstein Frisian (Ho) and Brown Swiss (BS) bulls born and maintained under tropical conditions. Characteristics evaluated every 2 weeks, from 27 to 63 weeks of age, included live weight, scrotal circumference, testicular diameter, semen quality and sexual behavior. Puberty was defined as the age at which a bull first produced an ejaculate containing at least 50 x 106 spermatozoa, with a minimum of 10% progressive motility. Testicular growth was linear in Ho bulls and quadratic in BS bulls. There was no breed difference in age at puberty (Ho, 333 +/- 15.8 days; BS, 311 +/- 10.5 days). However, at puberty, live weight and scrotal circumference tended to be greater in Ho (276 +/- 16.9 kg and 28.4 +/- 1 cm, respectively) than in BS bulls (233 +/- 11.3 kg and 25.9 +/- 0.7 cm, respectively), and testicular diameter was larger for Ho (5.5 +/- 0.24 cm) than for BS bulls (4.8 +/- 0.16 cm). Pooled data for all bulls for semen characteristics at puberty were: volume, 6.3 +/- 0.6 ml; progressive motility, 26.8 +/- 4.4%; sperm concentration, 58.5 +/- 13.9 x 10(6) spermatozoa/ml, and 351.5 +/- 91.2 x 10(6) spermatozoa/ejaculate. These values improved until at least 18 weeks after puberty. Eighty-five percent of bulls mounted heifers by 206 days of age, but only a few bulls had mounts with ejaculation during the study. It was concluded that reproductive development was similar between Ho and BS bulls, but slower than that reported for dairy bulls in temperate areas. Variation in some characteristics, such as scrotal circumference, was observed among bulls within each breed group, which might be of benefit for genetic selection.     

Effects of feeding treatment on growth rates,metabolic profiles and age at puberty,and their relationships in dairy heifers

Puberty attainment in dairy heifers has been widely studied from a hormonal point of view, but few studies have focussed on puberty–blood profile relationships during growth. We led experiments to determine the effects of feeding treatments on growth parameters, age at puberty and plasma biochemical profiles, and the relationships between age at puberty and metabolic profiles at 6, 9, 12 and 15 months (mo) of age. Blood samples were collected from 67 Holstein heifers, born between September 2011 and February 2012, every 10 days from 5.5 mo of age until heifers were considered pubertal (plasma progesterone concentration greater than 1.0 ng/ml) or oestrus synchronisation (November 2012; 11 to 15 mo of age). Heifers born before 30 November were fed either a standard diet (SD, n=27) or an intensive-plane diet (ID1, n=27) from 0 to 6 mo of age. This strategy aimed to reach 190 to 200 kg (SD) or 220 to 230 kg (ID1) BW at 6 mo of age. All heifers born after 1 December received an intensive-plane diet (ID2, n=13) from birth until oestrus synchronisation, in order to reach a similar BW at first insemination as heifers born before 1 December. Only 56 heifers reached puberty before oestrus synchronisation, at an average age of 10.3±2.2 mo (6.2 to 14.4 mo) and a BW of 296±40 kg (224 to 369 kg). There was no difference among the three feeding treatments until 6 mo, but at 9, 12 and 15 mo of age, ID2 (n=11) heifers weighed 37, 52 and 30 kg more than SD (n=22) and ID1 (n=23) heifers (P<0.001), respectively. Glucose, non-esterified fatty acids, albumin, aspartate aminotransferase, alkaline phosphatase and tartrate-resistant acid phosphatase, calcium, phosphorus, potassium and iron decreased with age, whereas β-hydroxybutyric acid, total cholesterol, creatinine, the creatinine : albumin ratio, alanine aminotransferase and chloride increased. The feeding treatment significantly affected creatinine, the creatinine : albumin ratio, and phosphorus and sodium levels, which were higher for ID2 heifers compared with SD and ID1. A logistic regression based on plasma metabolites at 6 mo of age to explain puberty attainment before or at 12 mo of age showed a positive relationship with plasma cholesterol (odds ratio=9.05). In conclusion, the feeding treatment had minor consequences on plasma metabolites, but it did affect growth performance.     

Treating heifers with GnRH from 4 to 8 weeks of age advanced growth and the age at puberty

In heifer calves, an early transient increase in circulating concentrations of LH is associated with early follicular development and is thought to regulate the timing of puberty. In an attempt to hasten the onset of sexual maturity, the early rise in LH concentration was advanced by injecting heifer calves with 120 ng/kg of GnRH (n=6) twice daily from 4 to 8 weeks of age; control calves received saline (n=6). Blood samples were collected every 15 min for 10h at 4, 8, 14, 20, 26, 32, 38, 44 and 50 weeks of age. Treatment with GnRH increased mean circulating concentrations of LH at 8 weeks of age (P<0.05), LH pulse frequency at 4 and 8 weeks of age (P<0.05), and reduced the mean age at puberty by 6 weeks (56.8+/-1.7 versus 62.8+/-2.4 weeks of age, for GnRH treated and control calves, respectively; P=0.04). Body weight gain was greater in GnRH-treated calves than control calves (P<0.05), and the rate of weight gain was shown to be a significant covariate within age at puberty. In conclusion, we suggest that the timing of the early rise in LH concentrations is a critical signal involved in the timing of puberty in heifers.     

Influence of Restricted Suckling and Level of Feed Supplementation on Postpartum Reproductive Performance of Zebu and Crossbred Cattle in the Semi-Arid Tropics

This study was carried out in central Tanzania on a group of 45 Zebu and 37 crossbred cows which were 4 to 10 years old. At calving time, the animals were allocated to one of the 4 treatment groups. In addition to free access to grazing for all cows in the study, in group H: AR (n = 18), cows were fed a high level of concentrate supplementation (4kg/day) and calves were artificially reared; in group H:RS (n = 24), cows were fed a high level of concentrate supplementation (4kg/day) and calves were only allowed restricted suckling up until the weaning age of 6 months. In group L:AR (n = 23) cows were fed a low level of concentrate supplementation (2kg/day) and calves were artificially reared; and in group L:RS (n = 17) cows were fed a low level of concentrate supplementation (2kg/day) and calves were only allowed restricted suckling up until the weaning age of 6 months. Milk progesterone was used as a means of determining the postpartum resumption interval (PRI) and the interval from parturition to conception (PCI). The overall PRI was 47.4 ± 0.4 days and was significantly affected by breed but not by calving season, with crossbred cows exhibiting a shorter PRI than Zebu cows. The effect of the treatments was significant, with cows in the group H:AR displaying a significantly shorter PRI than those in the other groups, while cows in group L:RS showed a significantly longer PRI than those in the other groups. The overall PCI was 149.5 ±3.7 days, and was not significantly affected by breed or calving season. The effect of the treatments was significant, with cows in the group H:AR having a significantly shorter PCI than cows in the other groups, while cows in group L:RS showed a significantly longer PCI than those in the other groups. Crossbred cows had higher live weights at calving (299.4 kg) than Zebu cows (272.6 kg), while all cows gained weight during the first 3 months after calving. The treatments had a significant effect on weight gain, with cows in the group H:AR gaining significantly more weight than those in the other groups. Cows which had high live weights at calving exhibited significantly shorter PRI and PCI than the lighter cows. Animals which had gained more than 5 kg during the first month after calving, or which had gained more than 8 kg during the first 3 months after calving, showed significantly shorter PRI and PCI than cows which had gained less weight. The results show that the calf rearing system and the level of feed supplementation interact with each other and can influence the postpartum anoestrous period in Zebu and Zebu crossbred cattle. Increasing the level of nutrition in restricted suckling cows tended to improve the postpartum anoestrous period, but the positive effects of supplementation could not completely compensate for the negative effects of suckling.     

Predictive value of palpation per rectum for detection of the CL in Zebu cattle as evaluated by progesterone concentrations and ultrasonography

The main objective of this study was to calculate the predictive value of the positive and negative diagnosis of the presence of a corpus luteum by palpation per rectum in Zebu cattle. Gyr breed (n = 16) heifers were palpated by a veterinarian every 3 d during March and April, and every 5 days during May. The presence or absence of a CL was recorded for 442 examinations. A blood sample was obtained after each examination, and a functional CL was considered to be present if plasma progesterone concentrations were above 1 ng/ml. Progesterone results were used as the reference to calculate the sensitivity and specificity of palpation per rectum for the diagnosis of a functional CL on different months. Predictive values for the positive and negative diagnoses were calculated using different hypothetical rates of the prevalence of CL. To evaluate the morphological basis of an erroneous diagnosis, ultrasonographic scanning of the ovaries was performed in 264 cases and the physical presence or absence of a CL was registered. The calculations were repeated using the ultrasonographic findings as the reference to evaluate the sensitivity and specificity of the diagnosis of a structural CL by per rectum palpation. Several heifers were not cyclic in March but began to cycle during the study. Thus, the prevalence of both functional and structural CL increased with time. As a result, the proportion of palpation diagnoses classified as true positives increased in May, while the proportion of true negatives decreased. Although the values of sensitivity and specificity remained constant from month to month, the predictive values changed markedly in May, when the predictive value of the positive test increased while the predictive value of the negative test decreased. These changes were due to differences in the prevalence of a CL and not to differences in the palpable characteristics of CLs in different months. The prevalence of a structural CL was always higher than that of a functional CL. The Cohen's Kappa test for concordance revealed better correspondence between palpation per rectum and ultrasound (k = 0.82) than between progesterone and either ultrasound (k = 0.68) or palpation (k = 0.66). These results show that a large proportion of the errors imputed to deficient palpation when progesterone concentrations are used as a reference are in fact the result of lack of correspondence between the physical and functional presence of a CL.     

Evaluation of crosses of Holstein, Jersey or Brown Swiss sires x Holstein-Friesian/Gir dams. 2. Female liveweights

The liveweights of 100 females sired by Holstein (H), Jersey (J) or Brown Swiss (BS) bulls out of Holstein-Friesian x Gir dams of 1/2 to 3/4 Holstein-Friesian fraction were compared. The animals were kept in a single herd under the same management. The data were analyzed separately for four age categories by least squares techniques. The models for cows and heifers older than 18 months included the fixed effects of breed of sire, Bos taurus fraction, season of weighing, days in milk class (for cows only), pregnancy status class, year and the random effect of animal nested within breed of sire x B. taurus fraction subclasses. The models for younger heifers included the same effects except for lactation and pregnancy status. Based on 2937 observations, the mean weights for 0- to 6-month-old calves for the groups with H, J or BS sires were 88 +/- 1, 77 +/- 1 and 75 +/- 1 kg, respectively, the mean weights for 1-year-old heifers were 168 +/- 2, 159 +/- 2 and 155 +/- 1 kg, for 2-year-old heifers, 327 +/- 4, 303 +/- 4 and 319 +/- 4 kg, for 3-year-old heifers, 380 +/- 5, 369 +/- 8 and 390 +/- 4 kg and for cows, 464 +/- 3, 413 +/- 2 and 478 +/- 2 kg. Cubic growth curves, which were different in the three sire breed groups, satisfactorily explained the cow weight changes with age (R(2)>or=0.98). Maximum weight was attained at 7.8, 9.5 and 9.2 years in cows with H, J or BS sires. Although breed of sire effects were not significant in heifers (P>0.05), those with H sires were the heaviest up to two years of age. The females with BS or J sires had similar weights up to one year of age, but thereafter the former reached similar weights as the females with H sires and declined less after attaining the maximum weight. The cows with J sires were the lightest at all ages. Since previous results showed similar protein and fat yields per day of calving interval in the three breed of sire groups, it is suggested that the lighter J crosses may be more economic than the other groups on account of their likely lower feed maintenance costs.     

Dietary protein or energy restriction influences age and weight at puberty in ewe lambs

The working hypothesis was that restricted protein content in diets of prepubertal ewe lambs would delay time of onset of puberty to a greater extent than restricted energy content. Restriction of dietary protein (metabolizable protein, MP) or energy (ME) intake on age and weight at puberty was studied in ewe lambs in two experiments. In Experiment 1, 85 crossbred ewe lambs were assigned by weight to one of five dietary treatment groups: control (CTL; 18 mg ME day⁻¹ and 173 g MP day⁻¹); energy restricted (ER; 10.2 mg ME day⁻¹ and 96 g MP day⁻¹); protein restricted (PR; 11 mg ME day⁻¹ and 66 g MP day⁻¹); ER for 9 weeks then switched to CTL (ER/CTL); PR for 9 weeks then switched to CTL (PR/CTL). Ewe lambs were weighed biweekly, bled weekly, and serum was assayed for progesterone to determine time of puberty. Average age at puberty differed among lambs fed the various diets (P < 0.01). Ewe lambs in the CTL group were younger and weighed more at puberty. Lambs fed diets restricted in protein or energy content were similar in age at puberty. Dietary switching, ER/CTL and PR/CTL, affected body weight and gain (P < 0.05), but did not result in changed age at puberty compared with lambs fed ER or PR diets. In Experiment 2, two breed types (maternal type and meat type) were used to compare responses of dietary protein restriction to dietary energy restriction. In Experiment 2, lambs (n = 60) were assigned to either the CTL, PR or ER diets. Lambs in the CTL group gained more weight than lambs fed the PR diet and lambs fed the PR diet gained more weight than lambs fed the ER diet (P < 0.05). Diet affected age (P < 0.001), but not weight at puberty (P = 0.13). Dietary energy and protein restriction influenced age at puberty in ewe lambs with energy restriction having a greater influence on delaying onset of puberty than protein restriction. Realimentation did not overcome effects of previous feed restriction by inducing an earlier age at puberty than ewe lambs fed the ER or PR diet throughout the experimental period. Body weight gain prepuberty is more important than type of dietary restriction (protein or energy) with regard to impacting age at puberty. We, therefore, reject the working hypothesis that dietary protein restriction prepuberty would have a greater impact in delaying onset of puberty than dietary energy restriction.     

Superstimulation of ovarian follicular growth with FSH oocyte recovery, and embryo production from Zebu (Bos indicus) calves: effects of treatment with a GnRH agonist or antagonist

The capacity of heifer calves of a late sexually maturing Zebu (Bos indicus) genotype to respond to superstimulation with FSH at a young age and in vitro oocyte development were examined. Some calves were treated with a GnRH agonist (deslorelin) or antagonist (cetrorelix) to determine whether altering plasma concentrations of LH would influence follicular responses to FSH and oocyte developmental competency. Brahman calves (3-mo-old; 140 +/- 3 kg) were randomly assigned to 3 groups: control (n = 10); deslorelin treatment from Day -8 to 3 (n = 10); and cetrorelix treatment from Day -3 to 2 (n = 10). All calves were stimulated with FSH from Day 0 to 2, and were ovariectomized on Day 3 to determine follicular responses to FSH and to recover oocytes for in vitro procedures. Before treatment with FSH, heifers receiving deslorelin had greater (P < 0.001) plasma LH (0.30 +/- 0.01 ng/ml) than control heifers (0.17 +/- 0.02 ng/ml), while plasma LH was reduced (P < 0.05) in heifers treated with cetrorelix (0.13 +/- 0.01 ng/ml). Control heifers had a surge release of LH during treatment with FSH, but this did not occur in heifers treated with deslorelin or cetrorelix. All heifers had large numbers of follicles > or = 2 mm (approximately 60 follicles) after superstimulation with FSH, and there were no differences (P > 0.10) between groups. Total numbers of oocytes recovered and cultured also did not differ (P > 0.05) for control heifers and heifers treated with deslorelin or cetrorelix. Fertilization and cleavage rates were similar for the 3 groups, and developmental rates to blastocysts were also similar. Zebu heifers respond well to superstimulation with FSH at a young age, and their oocytes are developmentally competent.     

Incidence of delayed ovulation in Holstein heifers and its effects on fertility and early luteal function

In the first of 3 experiments 134 first-service and 108 repeat-breeder Holstein heifers were palpated at 12-hour intervals starting 24 hours after insemination to compare the incidence of delayed ovulation in the 2 groups. Delayed ovulation was defined as failure to ovulate within the first 24 hours after insemination. Ovulation occurred within 24 hours post insemination in 92.1% of the animals and was delayed in 7.9% of the cases, with no differences between first-service and repeat-breeder heifers, indicating that the subfertility of the repeat-breeder animals was not due to delayed ovulation. The duration of the delay was at most 12 hours since all the animals had ovulated by 36 hours post insemination. Conception rate of the 19 animals with delayed ovulation (42.1%) was not different (P>0.05) from that of the 223 heifers that ovulated on time (44.8%). In a second experiment, no differences were detected between 15 heifers with delayed ovulation and 15 animals that ovulated on time with respect to their progesterone concentrations during the first 8 days post insemination, indicating that delayed ovulation is not associated with delayed luteinization or subnormal early luteal function. In the third experiment, the conception rate of 126 repeat-breeder heifers that were treated with hCG at the time of insemination was 26.7%; the conception rate of 101 repeat-breeder heifers that were inseminated twice, at 12 and 24 hours after the onset of estrus, was 34.6%; and the conception rate of 105 repeat-breeder heifers which were not treated with hCG and which were inseminated only once was 30.5% (P>0.05) It is concluded that delayed ovulation is not an important cause of infertility and does not constitute an important component of the repeat-breeding syndrome in Holstein heifers.     

Induction of twinning in crossbred heifers by ipsilateral frozen embryo transfer

Induction of twinning by means of nonsurgical ipsilateral transfer of two frozen embryos or of demi-embryos was attempted in 297 virgin crossbred dairy heifers that were kept at the following locations: in stables at our breeding station (Group 1), in a mountain field (Group 2) and at a private farm (Group 3). Of these, 187 heifers (63%) were diagnosed pregnant at 25 d after embryo transfer and 134 heifers (45%) were diagnosed pregnant at 60 d after transfer by ultrasonic echography or by palpation per rectum. At calving, 118 heifers produced 46 sets of twins and 72 single calves, for a total of 164 calves sixteen pregnant heifers aborted fetuses between 2 and 7 mo. The pregnancy rate of Group 1 (39%) was lower than that of Group 2 (48%) or Group 3 (50%). Abortion and mortality rates for Group 3 (8% and 8%, respectively) were significantly lower (P<0.05) than for Group 1 (16% and 12%) or Group 2 (13% and 18%). Twin calves had lower birth weights (P<0.05) than singles, but at 270 to 330 d of age, there was no significant difference between the weights of single and twin calves. Dystocia or difficult delivery was not observed heifers producing twins. Retention of the placenta was observed in twin calving heifers, with placenta retention levels the highest in Group 2 (33%) than followed by Group 1 (14%) and Group 3 (10%). When gestation periods were compared between single and twin calves in Groups 1 and 2, the gestation period of single calves was 5.0 to 7.0 d shorter than that of twin calves. No significant difference was observed between the gestation periods of single and twin births in Group 3.     

Effect of dietary energy and protein density on body composition,attainment of puberty,and ovarian follicular dynamics in dairy heifers

The objectives were to examine the effects of dietary energy and protein density on age and body composition at puberty, and on ovarian follicular dynamics during the pre- and peripubertal periods in Holstein heifers. In Phase 1, heifers were randomly allotted (n=10 per diet) at 100 kg body weight (BW) to diets with either low (P1L), medium (P1M) or high (P1H) energy and protein formulated for an average daily gain (ADG) of 0.5, 0.8 or 1.1 kg per day, respectively. During Phase 2 (P2), all heifers were fed ad libitum a common diet formulated for an ADG of 0.8 kg per day. Half the animals within the high (n=5) and low groups (n=5) entered P2 either at 12 months of age (P2H-12; P2L-12) or at 330 kg BW (P2H-330; P2L-330). Heifers fed P1H, P1M, P1L, and P2L-12 diets attained puberty at approximately 9, 11, 16, and 14 months of age, respectively (P<0.01). Urea space estimates of body fat and protein percent, and back-fat thickness, were lower in P1L heifers compared to P1H or P1M heifers at similar chronological ages (P<0.05) but did not differ at puberty (P>0.10). Compared to P1L heifers, P1H heifers had high amplitude LH pulses at 8 months, and high frequency low amplitude LH pulses at 10 months of age (P<0.05). The mean diameter (mm) of the dominant follicle was smaller (P<0.05) in P1L heifers (10.6) compared to P1H (12.8) or P1M (12.2) heifers at 8 months. Maximum size and growth rate of the nonovulatory dominant follicle increased with age (P<0.05) but did not differ between P1H and P1M heifers at puberty. The diameter (mm) of the nonovulatory dominant follicle, and the first and second ovulatory follicles were larger in P2L-12 heifers (14.0, 14.7, and 14.9) compared to P1M heifers (13.1, 12.5, and 11.9), while the peak progesterone levels and CL growth were lower (P<0.05) in the first cycle. In conclusion, dairy heifers attained puberty at a constant body weight and body composition independent of dietary manipulation, the size of dominant follicles increased with age in association with increased LH support, and heifers realimented from a low energy diet developed larger first ovulatory follicles and smaller CL with lower peak progesterone concentrations in the first cycle.     

Quantitation of glucocorticoid receptors in bovine skeletal muscle: Topographical distribution, sex effect and breed comparisons

The concentration of the cytosolic glucocorticoid receptor (GR) was determined in skeletal muscles of calves in order to study possible differences in individual muscles from different parts of the body as well as the influence of sex and breed. In male and female Simmental calves the topographical distribution of GR was similar: the lowest concentrations were seen in abdominal muscle, whereas in neck, shoulder and hindleg the GR concentrations were higher; this difference was more pronounced in male than in female calves. In general, female calves had about 2-fold higher GR concentrations than males. The cytosolic cortisol concentrations were differing neither between individual muscles nor between sexes. The cortisol secretion during a 24-h sampling period 1 week prior to slaughter showed no sex difference. GR concentrations in neck muscle of female calves of four different German cattle breeds (Holstein Friesian, Brown Swiss, Simmental and German Gelbvieh) were rather similar; however, when Brown Swiss with the highest GR levels were compared to Holstein Friesian calves with the lowest concentrations, a significant difference was evident (P < 0.05).