Modulation of luteinizing hormone and follicle-stimulating hormone in circulation by interactions between endogenous opioids and oestradiol during the peripubertal period of heifers.

The aim of this study was to determine whether the decline in oestradiol inhibition of circulating luteinizing hormone (LH) and follicle-stimulating hormone (FSH) during the peripubertal period of heifers is associated with a change in opioid modulation of LH and FSH secretion. Opioid inhibition of LH secretion was determined by response to administration of the opioid antagonist naloxone. Prepubertal heifers (403 days old) were left as intact controls, ovariectomized or ovariectomized and chronically administered oestradiol. Control heifers were used to determine time of puberty. Three weeks after ovariectomy, four doses of naloxone (0.13-0.75 mg kg-1 body weight) or saline were administered to heifers in the treatment groups in a latin square design (one dose per day). Blood samples were collected at intervals of 10 min for 2 h before and 2 h after administration of naloxone. This procedure was repeated four times at intervals of 3 weeks during the time intact control heifers were attaining puberty. All doses of naloxone induced a similar increase in concentration of serum LH within a bleeding period. During the initial bleeding period (before puberty in control heifers), administration of naloxone induced an increase in LH concentration, but the response was greater for heifers in the ovariectomized and oestradiol treated than in the ovariectomized group. At the end of the study when control heifers had attained puberty (high concentrations of progesterone indicated corpus luteum function), only heifers in the ovariectomized and oestradiol treated group responded to naloxone. Opioid inhibition of LH appeared to decline in heifers during the time control heifers were attaining puberty. Heifers in the ovariectomized group responded to naloxone at the time of administration with an increase in FSH, but FSH did not respond to naloxone at any other time. Administration of naloxone did not alter secretion of FSH in ovariectomized heifers. These results suggest that opioid neuropeptides and oestradiol are involved in regulating circulating concentrations of LH and possibly FSH during the peripubertal period. Opioid inhibition of gonadotrophin secretion appeared to decline during the peripubertal period but was still present in ovariectomized heifers treated with oestradiol after the time when age-matched control heifers had attained puberty. We conclude that opioid inhibition is important in regulating LH and FSH in circulation in heifers during the peripubertal period. However, opioids continue to be involved in regulation of circulating concentrations of LH after puberty.     

Effect of horn fly (Haematobia irritans) control on growth and reproduction of beef heifers

The influence of horn fly control with commercially available ear tags was studied on beef replacement heifers (n = 670) for growth and reproductive performance. The study was conducted at five sites in Louisiana over 3 yr. Heifers used were yearling replacement females that were exposed to fertile bulls during a limited spring breeding season that coincided with the horn fly season. In mid to late May of each year, heifers were randomly assigned to one of two horn fly treatments: untreated and treated for horn fly control. The trial continued each year until September or October at the end of fly season. Pregnancy status was determined by rectal palpation. Horn fly populations were controlled on the treated heifers at moderate levels (84%). Total weight gain of treated heifers was 14% greater than for untreated heifers. Horn fly treatment had no effect on pregnancy rate (78% and 75% for untreated and treated heifers, respectively). Treatment differences for weight gain were of greater magnitude for heifers failing to conceive than for heifers that became pregnant. Weight gains of nonpregnant treated heifers were 33% greater than for nonpregnant untreated heifers, whereas weight gains of pregnant treated heifers were 8% greater than for pregnant untreated heifers. In conclusion, horn fly control on yearling beef replacement heifers improved weight gain but had no effect on first exposure reproduction.     

Effects of dry season nutritional supplementation on growth, onset of puberty and subsequent fertility in Boran and Boran x Friesian heifers in Ethiopia

Shortly after weaning at 8 months of age, 43 Boran and 46 Boran x Friesian crossbred heifers were randomly divided to either receive a supplementary feed containing 16% crude protein during the dry season or to serve as controls. Heifers were examined monthly, and data on body weight, wither height and body condition score were recorded. Ovarian size and structures were determined per rectum and heifers were kept under continuous observation for standing estrus. Blood samples were collected at 10-day intervals every month for determination of plasma progesterone levels. All heifers were exposed to intact bulls for 4 months after they attained 18 months of age. Average daily weight gain to puberty was significantly (P<0.01) higher in heifers given supplementation than in the controls (360 vs 326 g/day). The average daily weight gain in Boran heifers (296 g/day) was significantly (P<0.01) lower than in crossbred (392 g/day) heifers. Mean age at puberty differed significantly (P<0.001) between heifers given supplementation (573 days) and the controls (627 days). Boran heifers attained puberty at a significantly later age (660 vs 540 days; P<0.001) than crossbred heifers. Body weight at puberty did not differ between supplemented and control heifers (226 vs 222 kg); while the difference between Boran and crossbred heifers was significant (216 vs 232 kg; P<0.01). Pelvic size and body condition score were not influenced by supplementation but differed significantly between genotypes. Pregnancy rate after 4 months of breeding was higher in supplemented (79%) than in control (64%) heifers. Boran heifers had a significantly (P<0.001) lower pregnancy rate than crossbred heifers (52 vs 91%). Mean age and body weight at conception were not affected by supplementation but differed significantly (P<0.05) between genotypes. The results indicate that prepubertal supplementary feeding during the dry season increased growth rate, reduced age at puberty and improved fertility in both Boran and Boran x Friesian heifers.     

Metabolic cues for puberty onset in free grazing Holstein heifers naturally infected with nematodes

Leptin is a new plausible candidate for the molecular link between nutritional status and the reproductive axis. In previous studies we described that continuous natural nematode infections in heifers retarded growth and delayed the onset of puberty, and that the insulin-like growth factor I (IGF-I) was involved. In the present study we monitored the leptin levels during development in heifers naturally parasitized versus those chronically treated with ivermectin and we investigated whether growth hormone (GH) accounted for the differences in IGF-I previously noted. Insulin levels were also measured. Prolactin hormone was recorded as an indicator of immune system activation. We found a direct correlation between leptin and body weight during development and a prepubertal surge of the hormone 2 weeks before the first progesterone peak that indicates the onset of puberty. This suggests that leptin may act as a signal for this event. Insulin did not vary during growth and prepuberty. On the other hand, GH as not responsible for diminished IGF-I levels in parasitized animals as levels were similar in both groups. The GH levels were high at birth and then diminished rapidly and remained constant during development and puberty. The last hormone studied, prolactin, followed seasonal changes of sunlight duration and presented sporadic bursts in infected animals. These were related to high nematode infection and are probably involved in the immune response of the host.     

Effect of level of nutrition on onset of puberty and conception rates of zebu heifers

Sixty zebu heifers were divided into three equal groups and reared on isocaloric diets but on different levels of protein. The protein levels were 19.17% (high), 13.37% (medium; NRC recommendations) and 8.3% (low). All animals were examined weekly per rectum for the presence of follicles and corpora lutea and also checked daily for standing heat. The presence of a mature corpus luteum was considered to indicate the attainment of puberty. Body weight and pelvic dimensions were taken at biweekly intervals. Intact bulls were introduced to heifers after they reached puberty and once they reached 200 kg live weight. Pregnancy diagnosis was carried out at 60, 90 and 120 days following introduction of bulls. Mean age at puberty for heifers in the high protein group (570.4 days), medium protein group (640.8 days) and low protein group (704.2 days) differed significantly (P<0.05). body weight at puberty for heifers in the high, medium and low protein groups was 207.1, 187.0 and 161.7 kg, respectively (P<0.05). Pelvic size at puberty was not influenced by the nutritional level. Conception rates of heifers in the three groups were influenced by level of nutrition. Pregnancy rates for the high protein group (58.8%), medium protein group (27.8%), and low protein group (16.7%) by 90-day post-breeding period were significantly different (P<0.05), but body weight at conception between the groups did not differ significantly. The results show that increasing the protein level in the diet is a means of improving the reproductive performance of zebu cattle.     

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.     

Delayed puberty and response to testosterone in a rat model of colitis

Delayed puberty is a frequent complication of inflammatory bowel disease. The precise etiological mechanisms are not known. In this study, we wanted to determine the relative contribution of undernutrition and inflammation to delayed puberty and the effect of inflammation on the reproductive axis. Puberty was assessed in rats with 2,4,6-trinitrobenzenesulfonic acid induced-colitis, healthy controls, and animals pair fed to match the food intake of the colitic group. The response to testosterone administration was assessed in colitic rats. We found that induction of colitis was associated with hypophagia and reduced weight gain, and undernutrition in healthy females (i.e., pair fed) resulted in a delay in the onset (by 4.8 days, P < 0.001) and progression of puberty (normal estrous cycles in 42%, P = 0.04) compared with controls. However, puberty was further delayed in the colitic group (1.4 days after pair fed) with the absence of normal estrous cycling in all rats. In males, the onset of puberty was also delayed, and weights of accessory sex organs were reduced compared with pair-fed controls. Plasma testosterone concentrations were low, and gonadotropin concentrations were normal in colitic rats. Testosterone treatment normalized puberty in male rats with colitis. In conclusion, in rats with experimental colitis, inflammation appears to potentiate the effect of undernutrition on puberty. The weights of secondary sex organs and the onset of puberty were normalized by testosterone treatment.     

Prenatal Testosterone Exposure Worsen the Reproductive Performance of Male Rat at Adulthood

The reproductive system is extremely susceptible to environmental insults, for example exogenous steroids during gestational development and differentiation. Experimental induction of androgen excess during prenatal life in female animal models reprograms their reproductive physiology, however the fetal programming of the male reproductive system by androgen excess has not been well studied. We aimed to determine the effect of prenatal exposure of two different doses of testosterone on different gestational days, on the male reproductive system using a rat model. Sixteen pregnant rats were randomly divided into two experimental groups and two control groups. Experimental group І were subcutaneously injected with 3 mg free testosterone on gestational days 16-19 and its controls received solvent for that time; experimental group П were subcutaneously injected with 20 mg free testosterone on day 20 of gestational period and its controls received solvent at the same time. The reproductive system morphology and function of 32 male offspring of these study groups were compared at days 6-30-60 of age and after puberty. The anogenital distance of the male offspring of both experimental groups had no significant differences on the different days of measurement, compared with controls. In the offspring of experimental group І, the testes weight, number of Sertoli, Spermatocyte and Spermatid cells, sperm count and motility and the serum concentration of testosterone after puberty were significantly decreased; except for reduction of sperm motility (p< 0.01), the other effects were not observed in the offspring of experimental group ІІ. In summary, our data show that prenatal exposure of male rat fetuses to excess testosterone disrupted reproductive function, an effect highly dependent on the time, duration and level of exposure. It seems that the reproductive system in individuals exposed to high levels of androgens during fetal life should be evaluated at puberty and likely to be treated.     

Genomic stability and physiological assessments of live offspring sired by a bull clone, Starbuck II

It appears that overt phenotypic abnormalities observed in some domestic animal clones are not transmitted to their progeny. The current study monitored Holstein heifers sired by a bull clone, Starbuck II, from weaning to puberty. Genomic stability was assessed by telomere length status and chromosomal analysis. Growth parameters, blood profiles, physical exams and reproductive parameters were assessed for 12 months (and compared to age-matched control heifers). Progeny sired by the clone bull did not differ (P>0.05) in weight, length and height compared to controls. However, progeny had lower heart rates (HR) (P=0.009), respiratory rates (RR) (P=0.007) and body temperature (P=0.03). Hematological profiles were within normal ranges and did not differ (P>0.05) between both groups. External and internal genitalia were normal and both groups reached puberty at expected ages. Progeny had two or three ovarian follicular waves per estrous cycle and serum progesterone concentrations were similar (P=0.99) to controls. Telomere lengths of sperm and blood cells from Starbuck II were not different (P>0.05) than those of non-cloned cattle; telomere lengths of progeny were not different (P>0.05) from age-matched controls. In addition, progeny had normal karyotypes in peripheral blood leukocytes compared to controls (89.1% versus 86.3% diploid, respectively). In summary, heifers sired by a bull clone had normal chromosomal stability, growth, physical, hematological and reproductive parameters, compared to normal heifers. Furthermore, they had moderate stress responses to routine handling and restraint.     

Estrus and pregnancy in beef heifers following use of the Syncro-Mate-B treatment (SMB)

Five studies were conducted in yearling heifers which were either puberal or prepuberal utilizing the Syncro-Mate-B (SMB) treatment (G. D. Searle & Co., Chicago, Ill.). In trial 1 the pregnancy rate in Brahman crossbred heifers after 21 days of breeding was 56% and 28% greater (P<.005) in the SMB treated groups than in the nontreated groups. Pregnancy rate in 3 4 Simmental heifers (trial 2) was increased 39% (P<.05) after 21 days of breeding. Hereford and crossbred heifers (trial 3) weighing less than 250 kg failed to respond to SMB treatment whereas heifers weighing greater then 250 kg had increased reproductive performance. Conversely the results obtained in trial 4 using Brahman crossbred heifers was not influenced by weight or SMB treatment. Crossbred Santa Gertrudis heifers (trial 5) weighing greater then 250 kg and treated with SMB had improved reproductive performance compared to the control groups. While in treated heifers weighing less than 250 kg reproductive performance was far from optimum.     

Effect of electrical stimulation or presence of a bull on puberty in beef heifers

Three experiments were conducted to determine if electrical stimulation or the presence of a bull would hasten puberty in Angus and Hereford heifers. In experiment I, electrical stimulation of the genital tract per rectum given once on day 0 of the experiment failed to hasten puberty and did not alter the size of large follicles. The presence of a teaser bull, for a period of three weeks, did not hasten puberty in a group of prepuberal heifers in experiment II. Experiment III was designed to determine whether a response to bulls could be discerned if heifers were treated with 12 mg progesterone daily for nine days. Introduction of teaser bulls to progesterone-treated heifers did not influence the percentage of heifers that reached puberty in either of two 30-day trials of this experiment. Progesterone alone did not significantly increase the percentage of heifers that reached puberty, but it appeared to synchronize estrus in heifers that reached puberty after treatment.     

Neonatal exposure to progesterone and estradiol alters uterine morphology and luminal protein content in adult beef heifers

Exposure of the developing urogenital tract to steroids can affect structure and function of adult tissues and compromise reproductive performance. This study was conducted to determine 1) if exposure of neonatal heifer calves to progesterone (P) and estradiol benzoate (E), delivered from a commercial growth-promoting implant, would affect adult uterine morphology or uterine luminal protein content; and 2) whether such effects would be related to neonatal age at the first exposure. At birth (Day 0), 20 crossbred beef heifers were assigned to 1 of 4 treatment groups (n = 5 per group), defined by age at implant placement. Heifers either received an implant on Days 0, 21 or 45, or served as untreated controls. The heifers were maintained together and slaughtered at 15 mo of age, during the luteal phase of an induced estrous cycle, when reproductive tracts and blood samples were obtained. Peripheral plasma P concentrations were determined by RIA. Uterocervical wet weights were recorded, and uterine luminal flushings (ULF) were assayed for total protein. Cross-sections of uterine tissues were evaluated histomorphometrically to determine myometrial and endometrial areas and relative endometrial gland density. Treatment did not affect plasma P concentrations (3.2 +/- 0.5 ng/ml). Regardless of age at treatment, neonatal PE exposure reduced uterocervical wet weight by 35% (112.8 < 173.9 +/- 13.9 g; P < 0.01), myometrial area by 23% (125.3 < 162.8 +/- 8.5 mm2; P < 0.02), and endometrial area by 27% (33.3 < 45.4 +/- 2.7 mm2; P < 0.09) compared with the untreated controls. Endometrial gland density was reduced (P < 0.01) by 40% in treated heifers. This effect was related to age at implant placement. Uterine gland density was reduced (P < 0.01) by 65% in heifers treated at birth, while reductions of 22 and 33% were observed for heifers treated on neonatal Day 21 or 45, respectively. Consistently, ULF protein content was lower (P < 0.01) in the treated heifers (2.67 < 4.98 +/-. 72 mg/ULF). Thus, exposure of newborn calves to PE can have profound effects on adult uterine morphology and environment, the extent of which may depend upon the developmental period when exposure occurs. The potential of such alterations to affect reproductive performance in adult beef heifers remains to be investigated.     

Pubertal traits and seasonal variation of the sexual activity in Brahman, Hereford and crossbred heifers

Age and weight at puberty and the pattern of ovulatory activity from puberty to 26 mo of age were studied in a total of 55 Brahman (B), Hereford (H), 5/8B-3/8H (B(5)H(3)) and 5/16B-11/16H (B(5)H(11)) heifers. The trial was conducted over two years. Heifers were kept during the whole experimental period on a paddock of native pasture at a stocking rate of 0.5 animals per hectare. Vasectomized bulls were used for estrus detection and blood was collected from the heifers twice weekly to monitor ovulatory activity through progesterone level as determinated by RIA. In addition to age and weight at puberty, the weights and dates at the time of cessation and resumption of ovulatory activity were also considered. There were significant effects (P<0.05 or P<0.01) of genetic group (purebreds vs crossbreds) for age and weight at puberty; the difference being 60 d and 18 kg, respectively, favoring the crossbred groups. After puberty attainment, an anestrous period was detected in all Brahman (118.6 d) and B(5)H(3) (113.4 d) heifers; anestrus was observed in only 14% of B(5)H(11) heifers and for a shorter (43.5 days) period. Hereford heifers did not exhibit interruption of ovulatory activity. In Brahman and B(5)H(3) heifers, the pattern of interruption and resumption of estrus and ovulatory activity correlated closely (0.89 and 0.95 for Brahman and B(5)H(3) heifers, respectively) with the photoperiod curve, suggesting the probable influence of this environmental factor on the regulation of estrous cycle activity in heifers with a higher proportion of Bos indicus genes. Thus, the genetic factor needs be taken into account when establishing programs in reproductive management.     

Endocrine mechanisms of puberty in heifers. Role of hypothalamo-pituitary estradiol receptors in the negative feedback of estradiol on luteinizing hormone secretion

The hypothesis tested was that the decline in negative feedback of estradiol on secretion of luteinizing hormone (LH) that occurs as puberty approaches in heifers results from a decline in the number of receptors for estradiol in the hypothalamus and/or pituitary. In addition, associated changes in receptors for luteinizing hormone-releasing hormone (LHRH) in the pituitary, ovarian follicle development, and uterine growth were characterized. Fifty prepubertal heifers, 234 to 264 days of age, were used. Six heifers of median body weight were designated controls, and sequential blood samples were collected at 20-min intervals for 24 h every 2 wk from 249 days of age through puberty and analyzed for concentrations of LH. Frequency of LH pulses/24 h was regressed on number of days prepuberty to develop a prediction equation for puberty. Thirty of the remaining 44 heifers were killed at 253, 302, and 351 days of age (n = 10/group), and tissues for described analyses were collected. Three to 5 days before tissue collection, sequential blood samples were obtained from these heifers, as described for control heifers to determine frequency of release of LH. With this information, number of days prepuberty at the time of tissue collection was estimated from the prediction equation developed with data from control heifers. The average age at puberty in control heifers was 366 days. The average age at puberty of heifers that were not killed or included in the control group (n = 14) was 360 days. Receptor and morphological data were related to the estimated onset of puberty. Cytosolic concentration of receptors for estradiol (fmoles receptor/mg cytosolic protein) in the anterior hypothalamus, medial basal hypothalamus, and anterior pituitary declined (p less than 0.05) as puberty approached. No change in concentration of receptors for estradiol was observed in the stalk median eminence or preoptic area. The concentration of receptors for LHRH in the anterior pituitary did not change as puberty approached. Uterine weight increased rapidly during the 50 days preceding puberty. The number of small, medium, or large follicles and the wet, pressed, or dry weight of the ovaries did not change as puberty approached. Follicles with a diameter greater than 12 mm were found only in the 3 heifers estimated to be closest to puberty at the time of tissue collection. The hypothesis that the decline in estradiol feedback on secretion of LH during the prepubertal period in heifers may result from a decline in the concentration of binding sites for estradiol at the hypothalamus and/or pituitary is supported by this study.     

Effect of selenium and vitamin E supplementation during the late pregnancy on reproductive indices and milk production in heifers

A trial was performed to determine the effects of supplementation of selenium (Se) and vitamin E (VE) on reproductive indices and milk production in Holstein heifers. Sixty heifers at the late stage of gestation were randomly assigned into three groups. Heifers were balanced for age, weight and time of calving. Four and 2 weeks before expected calving the heifers were injected 0 ml (C), 20 ml (T1), and 40 ml (T2) Se and VE supplements. Each ml contained 0.5mg Se and 50 IU of D, L-alpha-tocopheryl acetate. Blood samples were collected from heifers 4 weeks before expected calving and at calving day. Se concentrations in serum and colostrums were measured. The reproductive parameters were recorded. The effects of Se and VE supplements on somatic cell count (SCC) and milk yield also were measured. Supplementation of Se increased the level of Se in serum of treated heifers at calving day (P<0.05). Se concentration of colostrums were affected by the treatments (P<0.05). Concerning reproductive performance of treated heifers; gestation length was similar among groups but in comparison with controls, T1 and T2 had no incidence of retained fetal membrane (i.e., beyond 12h.). In addition, open days were fewer in T1 and T2 heifers and the number of services per conception was fewer in T2 compared with controls (P>0.05). Daily milk production at 8 weeks lactation was significantly increased in T2 compared with controls (P<0.05). Likewise, the milk SCC decreased in treated heifers compared with controls (P<0.05).     

Control of Boophilus annulatus (Acari: Ixodidae) on cattle using injectable microspheres containing ivermectin

The efficacy of an injectable microsphere formulation of ivermectin for control of the cattle tick, Boophilus annulatus (Say), was tested on 2 groups of 6 Hereford heifers held on separate 7-ha, tick-infested, buffel grass pastures. Cattle in one pasture were injected subcutaneously in the neck with a controlled-release microsphere formulation of ivermectin at the rate of 2.4 mg AI/kg body weight; the other group was injected with carrier only. Beginning 4 wk after injection and continuing throughout the remainder of the test (16 wk), no engorged ticks (> or = 5.5 mm) were found on any of the treated cattle, whereas large numbers of engorged ticks were found on the untreated controls. During this period, a few ticks were recovered from untreated sentinel animals placed in the treatment pasture during 7-8 wk after treatment, but none were recovered from animals exposed from 11-12 wk or 14-15 wk. Large numbers of B. annulatus ticks were found on untreated sentinel cattle placed in the control pasture during these same periods. Although the cattle, pastures, and tick habitat were approximately equal, the treated cattle gained an average of 77 kg compared with an average of 42 kg for the control group. This technology offers a possible alternative to the current official program of dipping and vacating pastures for eradication of Boophilus sp. infestations from the quarantine zone in southern Texas. Larger scale testing is needed to determine the potential of the injectable microsphere formulation and to optimize its use in eradication or control strategies.     

Reproductive response in supplemented heifers in the humid tropics of Costa Rica

To evaluate the effect of two supplementary diets to determine the consequence on productive and reproductive performance in heifers (Bos indicus x Bos taurus) averaging between 24 and 36 months of age and grazing tropical pastures, two trials were conducted. Thirty animals (initial BW 325.1+/-33.6 kg) were divided in two groups in the initial study: supplemented (SG) and control (CG); SG received a concentrate (5.5% CP and 2.85 Mcal/kg of DE dry matter basis) at 1% of body weight (BW). In the second study, 45 heifers (initial BW 332.6+/-29.3 kg) were assigned in two treatments, with the same amount of supplement (1% BW) but with a greater nutrient content (13% CP and 3.15 Mcal/kg of DE). The proportion of animals with a corpus luteum at the end of each study was greater in the supplemented groups (P<0.05). Ovarian follicular dynamics was similar between groups in the first study, but in the second study there were more heifers in the SG group with follicles larger than 9 mm in diameter (P<0.05). Pregnancy rate was similar for SG and CG (P>0.05). The response to a regimen of estrous synchronization in both trials was numerically superior in the SG group. No differences were observed in the length of estrus. Daily gain and body condition score were similar for supplemented and control groups (0.27 compared with -0.06 in the first study and 0.90 kg compared with 0.60 in the second study, respectively). Dietary supplementation improved the number of animals initiating estrous cycles and the expression of estrus when compared with unsupplemented control heifers. The dietary regimens imposed in these studies appear to be an adequate for the management of growing heifers destined to a reproductive program.     

Effect of supplemental light on growth,prolactin, progesterone and luteinizing hormone in water buffalo (Bubalus bubalis)

Fifty non-pregnant Surti buffalo heifers aged between 17 and 42 months (n=24, <24 months;n=26, >24 months) were randomly assigned to groups subject to either natural daylight +4h supplemental light (n=25) or natural day light (n=25), to study changes in growth, serum prolactin (Prl), progesterone (P₄) and luteinizing hormone (LH) to supplemental lighting. Ambient temperatures (T) and relative humidity (RH) generally were >27° C and <70% during the day-time, respectively. Light-supplemented heifers had 16.2 kg net body weight (BW) gain at 9 weeks compared to 20.8 kg for controls, but higher mean Prl after 6.5 weeks (P<0.01), and higher P₄ (0.41 vs 0.19 ng/ml;P<0.06) than control heifers. Older heifers had 39.7% greater BW (P<0.01), but a net 4.3% BW gain compared to a 10.1% gain for younger heifers at 10 weeks. Older, light-supplemented heifers had higher mean P₄ (0.63 vs 0.19 ng/ml;P<0.07) than the other groups. These weight and hormonal changes suggest that 4 h supplemental light can alter growth and endocrine function in buffaloes under similar planes of nutrition. While light supplementation did not have a positive effect on body wieght during the 10 week study, body weight and endocrine changes due to supplemental light may be important factors for initiation of reproductive cyclicity.     

Treatment with 17β-oestradiol does not influence age and weight at puberty in Bos indicus heifers

The working hypothesis was that treatment of heifers with 17β-oestradiol (E₂) during specific periods of prepuberty would reduce the response of the hypothalamic–pituitary axis to E₂ negative feedback and induce an earlier onset of puberty. The effects of chronic treatment with exogenous E₂ administered at specific maturational phases on the age and weight at puberty were studied in 96 prepubertal Brahman (3/4–7/8 Bos indicus) heifers (187.0±3.3 days of age, mean±SEM), weighing 149.9±2.5 kg. Heifers were randomly assigned to one of six groups (n=16 per group). Groups 2–6 received E₂ implants (Compudose 200®) for 90-day periods starting at 10, 13, 16, 19 and 22 months of age, while animals in group 1 remained untreated. Implants were placed subcutaneously at the base of the ear. Blood was collected for progesterone (P₄) determination by radioimmunoassay (RIA) and the animals were weighed at monthly intervals from 6 to 15 months then weekly from 15 to 28 months of age. Puberty was defined by concentrations of P₄>1 ng/ml in plasma and identification of a corpus luteum (CL) by transrectal ultrasonography (Aloka 210DX:7.5 MHZ probe). Treatment with exogenous E₂ at any of the ages/treatment intervals evaluated in this study did not reduce age or weight at puberty (P>0.7). The mean age and weight at puberty of control heifers was 735.3±19.7 days (range: 597–861) and 299.2±10.2 kg (range: 233–382), respectively, which is greater than the age and weight at puberty of 481 days and 246 kg, that was previously reported for B. indicus heifers [Post, T.B., Reich, M.M., 1980. Puberty in tropical breeds of heifers as monitored by plasma progesterone. Proceedings of the Australian Society of Animal Production 13, 61–62.]. The large variation in age and weight at puberty that was observed in the present study among heifers might indicate an individual animal effect to E₂ treatment among some of the treated animals. The lengthy interval from birth to puberty observed in this study, as compared to other studies, reflects the effects of other factors such as genotype, environmental or nutritional influences on puberty.     

Reproductive characteristics of cloned heifers derived from adult somatic cells.

This study examined the onset of puberty, follicular dynamics, reproductive hormone profiles, and ability to maintain pregnancy in cloned heifers produced by somatic cell nuclear transfer. Four adult somatic cell-cloned heifers, derived from a 13-yr-old Holstein cow, were compared to 4 individual age- and weight-matched heifers produced by artificial insemination (AI). From 7 to 9 mo of age, jugular venous blood samples were collected twice weekly, and from 10 to 11 or 12 mo of age, blood sampling was carried out every other day. After the heifers reached puberty (defined as the first of 3 consecutive blood samples with peripheral plasma progesterone concentrations of >1 ng/ml), ultrasound examination of ovaries and jugular plasma sample collection were carried out daily for 1 estrous cycle. Cloned heifers reached puberty later than controls (mean +/- SEM, 314.7 +/- 9.6 vs. 272 +/- 4.4 days and 336.7 +/- 13 vs. 302.8 +/- 4.5 kg for clones and controls, respectively; P < 0.05). However, cloned and control heifers were not different in estrous cycle length, ovulatory follicle diameter, number of follicular waves, or profiles of hormonal changes (LH, FSH, estradiol, and progesterone). Three of the 4 clones and all 4 control heifers became pregnant after AI. These results demonstrate that clones from an aged adult have normal reproductive development.