Comparison of different transvaginal ovum pick-up protocols to optimise oocyte retrieval and embryo production over a 10-week period in cows.

The objective was to develop a simple and effective ovum pick-up (OPU) protocol for cows, optimised for oocyte harvest and subsequent in vitro embryo production (IVP). Five protocols differing in collection frequency, dominant follicle removal (DFR) and FSH stimulation were tested on groups of three cows each, over an interval of 10 consecutive weeks. Performance was evaluated on per OPU session, per week and pooled (3 cowsx10weeks) basis. Among the non-stimulated groups, on a per cow per session basis, once- or twice-weekly OPU had no effect on the mean (+/- S.E.M.) number of follicles aspirated, oocytes retrieved and blastocysts produced (0.6+/-0.8 and 0.7 +/- 0.7, respectively). However, DFR 72 h prior to OPU almost doubled blastocyst production (1.2 +/- 1.3). In stimulated groups, FSH treatment (80 mg IM and 120 mg SC) was given once weekly prior to OPU. Treatment with FSH, followed by twice-weekly OPU, failed to show any synergistic effect of FSH and increased aspiration frequency. When FSH was given 36 h after DFR, followed by OPU 48 h later, more (P < 0.05) follicles (16.0 +/- 5.0), oocytes (10.6 +/- 4.5) and embryos (2.1 +/- 1.2) were obtained during each session, but not on a weekly basis. Pooled results over 10 weeks showed an overall improved performance for the treatment groups with twice-weekly OPU sessions, due to double the number of OPU sessions performed. However, the protocol that consisted of DFR, FSH treatment and a subsequent single OPU per week, was the most productive and cost-effective, with potential commercial appeal.

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.

Time estimation in Parkinson's disease: normal long duration estimation despite impaired short duration discrimination.

It has been claimed that patients with Parkinson's disease (PD) are deficient in estimating and reproducing time intervals in the range of seconds. This deficit is more severe when subjects are requested to count internally during the demanded intervals, and when the rate of internal counting is fast. The observed deficit might therefore reflect slow internal counting, i. e. motor slowness, rather than a specific deficit of temporal processing. However, PD patients also have a higher temporal discrimination threshold for sensory stimuli, a finding purportedly indicating a slow 'internal clock'. In this study we examined PD patients' processing of short durations (approx. 1s,'psychological present') and long durations (up to 48 s,'extended present'). In the first experiment the ability to discriminate between temporally overlapping presentations of two visual stimuli (darkened rectangles on the computer screen) in the range of one second was tested. In the second experiment PD patients' ability to estimate time intervals between 12 and 48 s was investigated. During these intervals, subjects were required to tap and read a random number on the computer screen at a rate of 1 Hz. We found that the patients were impaired at discriminating between short intervals. This deficit was independent of task difficulty and appeared to be based on an impairment of divided attention. Despite this deficit, the PD patients estimated time intervals up to 48 s as accurately as the controls. We suggest that time estimation, i. e. the feeling for the flow of time, is normal in PD patients despite impaired discrimination of brief intervals in the range of seconds.

Influence of somatotropin and nutrition on bovine oocyte retrieval and in vitro development.

The objective of this study was to determine the effects of supplemental bovine somatotropin (bST) and limit feeding on follicular growth and oocyte competence in yearling beef heifers. Sixteen growing heifers (424+/-4 kg) were randomly assigned to 1 of 4 treatments in a 2 x 2 factorial arrangement, with main effects of bST (0 or 33 microg/kg BW/d) and feeding regimen (ad libitum or 0.75 ad libitum intake). Animals were treated for 100 d prior to follicular aspiration, and treatments continued for the 42-d period that follicles were aspirated. Follicles were observed ultrasonically then aspirated, and recovered oocytes were matured, fertilized and developed in vitro. The number of follicles observed ultrasonically was greater with bST treatment (P<0.01) but was unchanged by plane of nutrition. The number and quality of recovered oocytes were similar among treatments, as was the number of oocytes resulting in blastocyst formation.

Developmental competence of juvenile calf oocytes in vitro and in vivo: influence of donor animal variation and repeated gonadotropin stimulation.

Juvenile calf oocytes represent an untapped source of germ plasm for reproduction. Reports on the developmental competence of calf oocytes have been controversial. In this research, oocytes were recovered after gonadotropin stimulation from Holstein calves (N = 10) at 2-3 mo of age (2-mo cycle) and again at 4-5 mo of age (4-mo cycle). The in vitro developmental competence was measured, and prestimulation follicle numbers (for 2-mo cycle) and poststimulation follicle numbers (both cycles) were obtained. The number of antral follicles doubled after stimulation (23.4 +/- 6.1 vs. 55.1 +/- 16.1) for the 2-mo cycle and for the 4-mo cycle (47.4 +/- 12.4). The number of follicles observed prior to stimulation in the 2-mo cycle was found to be highly correlated with the poststimulation oocyte recovery for both collection cycles (r = 0.95, 2-mo cycle; r = 0.81, 4-mo cycle). The majority (90-96%) of recovered oocytes were found to be usable for in vitro maturation and fertilization; of these, 41-42% cleaved and 10-11% developed to morulae or blastocysts. Eighty-four in vitro-produced embryos were transferred to synchronized recipients and resulted in 11 pregnancies, leading to 7 live (4 males, 3 females) and 2 dead (one male, one female) calves at full term. No significant differences were observed between the 2-mo and 4-mo collection cycles; however, 73% of the total pregnancies resulted from the 2-mo cycle. All pregnancies resulted from embryos of high-responding donors. The high correlation between the number of follicles prior to stimulation and the poststimulation response suggests the possibility of screening calves prior to stimulation for routine embryo production.

The influence of unilateral castration on testicular morphology and function in adult rams.

An experiment was designed to investigate the mechanisms controlling testicular compensatory hypertrophy in rams. Endocrine and histological events were examined, with special attention to Sertoli cell hyperplasia and hypertrophy as contributing factors to the compensatory process. Fifteen sexually mature yearling Targhee rams were allotted to intact control (C, n = 5) and unilateral castrate (UC, n = 10) treatment groups in June. Approximately 150 days after UC, testicular tissue was collected in November after efferent duct cannulation and rete testis fluid (RTF) collection or perfusion fixation. Unilateral castration increased mean testis weight by 56% (p = 0.01) and mean epididymal weight by 15% (p = 0.05). Although the mean volume of RTF collected more than doubled after UC (1.55 +/- 0.86 vs. 0.63 +/- 0.10 ml for UC and C rams, respectively), the difference was not statistically significant. By 150 days after UC, the concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) in jugular venous blood did not differ between the two treatment groups. The concentrations of T. dihydrotestosterone (DHT), and androgen-binding protein (ABP) in RTF were also similar for UC and C rams. However, since the observed mean RTF volume was increased, the amounts of T, DHT, and ABP exiting the testes of these UC rams via the RTF were approximately doubled, although this difference was not statistically significant. UC increased the mean diameter of seminiferous tubules by 21% (p less than 0.01) and of their lumina by 51% (p less than 0.01), but did not significantly increase mean height of seminiferous epithelium or estimated length of seminiferous tubules per testis.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of unilateral castration on morphologic characteristics of the testis in one-, two-, and three-year-old stallions.

The effects of unilateral castration on testicular compensatory hypertrophy were measured in 12 Morgan stallions, four each at one, two, and three years of age. They were randomized within age to intact (IN) or unilaterally castrated (UC) groups. Allotment and surgery were in January 1983 and total castration was in June 1983, 150 d after unilateral castration. Testis weight increased linearly with age (P < 0.01) and was increased by unilateral castration (P < 0.07). Epididymal weight also increased linearly with age (P < 0.05) and was heavier in UC animals (P = 0.15). Tubule diameter (P < 0.10) and epithelial height (P < 0.03) were greater in UC than in IN stallions. In conclusion, testes of stallions underwent compensatory hypertrophy after unilateral castration.

Effects of unilateral castration on serum luteinizing hormone, follicle stimulating hormone, and testosterone concentrations in one-, two-, and three-year-old stallions.

The endocrine control of compensatory hypertrophy was investigated in 12 Morgan stallions, four each at one, two and three years of age. Half were assigned to be unilaterally castrated (UC) in January and half to remain intact (IN). Nine blood samples were taken from each stallion at half-hour intervals 30, 90, and 150 d after unilateral castration for radioimmunoassay of serum concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), and testosterone. Mean serum LH concentration was greater (P<0.06) in UC than IN stallions; however, the difference was greatest at 30 d and least at 150 d. Serum LH was greater (P<0.01) in two- and three-year-olds than in one-year-olds. The mean log(10) for serum FSH concentration was greater (P<0.06) in UC than IN stallions. Mean serum testosterone concentrations were similar in UC and IN stallions for all sample days, suggesting that the single testes of the UC stallions produced as much testosterone as the two testes of the IN stallions. Two- and three-year-old stallions had greater (P<0.01) serum testosterone than one-year-old stallions. Unilateral castration of stallions was associated with a significant increase in serum LH and FSH concentrations and, perhaps, higher intratesticular testosterone, which may explain, in part, the compensatory hypertrophy noted in the remaining testis.

Factors influencing estrus and conception in dairy heifers after prostaglandin F2-alpha.

The influence of interval between insemination (AI) and estrus on subsequent fertility of PGF2alpha-treated (two injections of 25 mg, 11 days apart) heifers was assessed in two experiments. In Experiment I, 240 heifers were allotted to Control (AI 8 to 16 hr after estrus detection), PGF2alpha-E (AI 8 to 16 hr after estrus within five days of second PGF2alpha) or PGF2alpha-T (AI 80 hr after second PGF2alpha). In Experiment II, 130 heifers were assigned to control (AI as before) or PGF2alpha (AI 72 or 80 hr after second PGF2alpha) with half the PGF2alpha heifers receiving 100 microg GnRH 72 hr after first PGF2alpha. Heifers of both experiments that were bred at a predetermined time were arrayed by interval from AI to estrus. Conception rates of heifers detected in estrus from 32 hr before AI to 24 hr after AI did not differ (x2=3.35, df=5, P>0.5). The percentage of GnRH-treated heifers in estrus within five days (81.8%) was not (P>0.75) greater than those not receiving GnRH (77.3%) but they had higher (P<0.05) serum progesterone (P4) concentration at second PGF2alpha (3.17 vs 2.41 ng/ml). When P4 values were arrayed for both groups at 1 ng intervals, the percentage of heifers exhibiting estrus increased with increasing P4 level (P<0.05).

Regulation of the estrous cycle in ewes with progestin containing implants: Influence of dose and day of cycle at treatment.

Implants containing Norgestomet (G. D. Searle and Co., Chicago) were inserted subcutaneously in ewes on selected days of the estrous cycle. When ewes were treated for 13 days with 2 or 3 mg Norgestomet, implantation 13 days post-estrus reduced the number of ewes in estrus within 5 days of implant removal and reduced the number of estrous ewes that lambed compared with ewes implanted 4 days post-estrus. When ewes were implanted with 3 or 6 mg Norgestomet 4 or 13 days post-estrus, no difference in estrus response was found. Conception rate was not influenced by day of treatment, but was higher in those ewes treated with 6 mg than ewes treated with 3 mg. Compared to no treatment, treatment with 3 or 6 mg Norgestomet reduced the number of uterine and oviducal sperm recovered 12 or 24 hr after insemination from ewes implanted for 12 days 2 or 12 days post-estrus. However, more sperm were recovered from ewes treated 2 days than 12 days post-estrus with the principal increase occurring in ewes treated with 6 mg of Norgestomet.

Influence of unilateral castration and increased plane of nutrition on sexual development of Holstein bulls. I. Growth and sperm production.

Sixty-five Holstein bull calves were used to study the effects of unilateral castration (UC) and increased plane of nutrition on the growth and development of the reproductive system. Bulls were slaughtered at 1 wk., 2, 4, 8 and 16 months. Half of each slaughter group above one week was unilaterally castrated at 7 days of age. Half of the bulls remaining at 6 months of age received 90% of their recommended daily TDN allowance while the remainder received 120%. Compensatory hypertrophy was evident as early as 2 months and the degree of compensation increased for the duration of the experiment (Age x UC, P<.01). By 16 months of age the remaining testis of UC animals was 73% heavier than the average testis weight of intact bulls. While epididymal weight was significantly increased by UC, seminal vesicle weight was not. UC bulls produced significantly more sperm per testis than intact bulls both from the onset of puberty to slaughter and for the 16 week period prior to slaughter. Testis sperm concentration was similar in UC and intact bulls. UC at one weel of age caused greater testis growth and greater sperm production per testis, but did not promote earlier puberty.

Influence of unilateral castration and increased plane of nutrition on sexual development of Holstein bulls. II. Histologic development of the testes.

Sixty-five Holstein bull calves were assigned in an experiment to determine the effects of unilateral castration (UC) at 1 week of age and of two levels of nutrition after 6 months on reproductive development to 16 months. Five animals were killed at 1 wk and half the remainder UC at that age. Groups of 5 in all factorial groups were killed at 2, 4, 8 and 16 months. Tubular diameter increased with age (P<.01) and at 8 and 16 months with UC (P<.01). Epithelial area at 8 and 16 months increased with age and UC (P<.01). The percents of tubular and intertubular tissue varied with age (P<.01) with the tubular tissue having its highest value at 8 months. Indexes of both total tubular and intertubular tissue were increased with age and UC (P<.01). The number of type A spermatogonia per cross section of stage 1 tubules of 16-month bulls was increased by UC (P<.05).

Influence of unilateral castration and increased plane of nutrition on sexual development of Holstein bulls. III. Endocrine responses.

Pituitary gonadotropic hormones were assayed in 65 Holstein bulls from 7 days to 16 months. Pituitary LH concentration and content at 2, 4, 8 and 16 months increased (P<.01) with age, while FSH content increased with age (P<.01) but was lower in UC bulls at 2, 4 and 8 months and higher at 16 months (A x UC, P<.01) as compared to intact bulls. In five samples of plasma collected at 90-minute intervals, one day each month from 1 to 15 months in 10 of the bulls killed at 16 months, LH concentration and variance changed (P<.01) with age reaching maxima at 4 and 3 months respectively. Plasma testosterone concentration and variance changed (P<.01) with age, reaching maxima at 10 and 9 months respectively. In the other 10 bulls killed at 16 months, assays of plasma collected before and after exposure to a teaser showed that stimulation increased LH by 20 minutes after exposure but LH declined by 60 minutes while testosterone was increased 20 (P<.05) and 60 (P<.01) minutes after exposure.