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The effect of type of vaginal insert and dose of pLH on embryo production, following fixed-time AI in a progestin-based superstimulatory protocol in Nelore cattle.

The objective was to analyze and report field data focusing on the effect of type of progesterone-releasing vaginal insert and dose of pLH on embryo production, following a superstimulatory protocol involving fixed-time artificial insemination (FTAI) in Nelore cattle (Bos taurus indicus). Donor heifers and cows (n = 68; 136 superstimulations over 2 years) received an intravaginal, progesterone-releasing insert (CIDR or DIB, with 1.9 or 1.0 g progesterone, respectively) and 3-4 mg of estradiol benzoate (EB) i.m. at random stages of the estrous cycle. Five days later (designated Day 0), cattle were superstimulated with a total of 120-200 mg of pFSH (Folltropin-V), given twice daily in decreasing doses from Days 0 to 3. All cattle received two luteolytic doses of PGF2alpha at 08:00 and 20:00 h on Day 2 and progesterone inserts were removed at 20:00 h on Day 3 (36 h after the first PGF2alpha injection). Ovulation was induced with pLH (Lutropin-V, 12.5 or 25 mg, i.m.) at 08:00 h on Day 4 with FTAI 12, 24 and in several cases, 36 h later. Embryos were recovered on Days 11 or 12, graded and transferred to synchronous recipients. Overall, the mean (+/-S.E.M.) number of total ova/embryos (13.3 +/- 0.8) and viable embryos (9.4 +/- 0.6) and pregnancy rate (43.5%; 528/1213) did not differ among groups, but embryo viability rate (overall, 70.8%) was higher in donors with a DIB (72.3%) than a CIDR (68.3%, P = 0.007). In conclusion, the administration of pLH 12 h after progesterone removal in a progestin-based superstimulatory protocol facilitated fixed-time AI in Nelore donors, with embryo production, embryo viability and pregnancy rates after embryo transfer, comparable to published results where estrus detection and AI was done. Results suggested a possible alternative, which would eliminate the need for estrus detection in donors.

Administration, Intravaginal↗

Comparison of long-term feeding pattern between male and female Fischer 344 rats: influence of estrous cycle.

We studied the effect of gender on food intake, meal number, and meal size in eight 10-wk-old female and seven age-matched male Fischer 344 rats for 44 consecutive days. Although food intake (g/100 g body wt) was similar in males and females (5.42 +/- 0.10 vs. 5.13 +/- 0.13 g food.day-1.100 g body wt-1, respectively; not significant), weight gain in males was approximately seven times greater than in female rats (1.49 +/- 0.07 vs. 0.21 +/- 0.03 g/day, respectively; P < 0.001). During this time, males had a relatively constant food intake. They increased their meal size but decreased their meal number. In female rats, food intake was relatively stable for the duration of the study, despite cyclically and reciprocally recurring changes in meal number and meal size, which are synchronized with the estrous cycle. Data confirm that net food intake is a dynamic process and suggest that, in the rat, the homeostasis of food intake in response to external as well as internal stimuli is maintained via the modulation of meal number and size.

Animals↗

Waves of follicle development during the estrous cycle in sheep.

The pattern of ovarian follicle development in maiden cyclic lambs was characterized using the definition of a follicle wave as the changes in the number of follicles among the days of the estrous cycle, as originally defined in cattle by Rajakoski in 1960. We also examined the steroid content relationships among follicles on Days 5 (Wave 1) and 14 (Waves 2 and 3) of the estrous cycle. In Experiment 1, the ovaries of 20 cyclic lambs (40 to 45 kg) were examined daily using transrectal ultrasonography for 1 or 2 estrous cycles (n = 31 cycles). The number of small (2 and 3 mm in diameter), medium (4 and 5 mm) and large (> or = 6 mm) follicles were aligned with the beginning and end of the average length estrous cycle and then compared among days. Identified follicles were defined as those that grew to > or = 4 mm and remained at > or = 3 mm for > or = 3 d. The number of identified follicles emerging (retrospectively identified at 2 or 3 mm) per ewe per day was also aligned with the average length estrous cycle. In Experiment 2, ewe lambs were ovariectomized on Day 5 (n = 6) or 14 (n = 5) of the estrous cycle, then follicle diameters and follicular fluid concentrations of estradiol and progesterone were compared among follicles. Data were analyzed by repeated measures ANOVA and compared among days using Fisher's LSD. In Experiment 1, either 2 (n = 10 cycles), 3 (n = 20 cycles) or 4 (n = 1 cycle) periods of emergence of identified follicles occurred during individual cycles, with estrous cycle lengths of 15.6 +/- 1.6, 16.1 +/- 1.1 and 17 d respectively. In animals with 2 or 3 periods of emergence of identified follicles, the total number of small, medium and large follicles differed (P < 0.05) among days of the estrous cycle showing a wave-like pattern. In Experiment 2, a single follicle collected on each of Days 5 and 14 of the cycle (6.2 +/- 0.2 and 3.9 +/- 0.2 mm in diameter) had a higher (P < 0.05) concentration of follicular fluid estradiol (36.2 +/- 4.4 and 50.9 +/- 21.6 ng/mL) than other follicles collected on the same day (next largest follicle: 4.3 +/- 0.3 and 3.5 +/- 0.4 mm; 4.3 +/- 0.9 and 18.2 +/- 6.7 ng/mL estradiol). The results showed that 1) there was a synchronous emergence of follicles associated with fluctuations in the number and size of follicles during the estrous cycle; 2) within a wave there was a hierarchy among follicles for diameter and steroid content; 3) ovarian follicle growth in ewe lambs occurred in 2 or 3 organized waves during the estrous cycle.

Animals↗

Acetylcholine rhythm in the preoptic area of the rat hypothalamus is synchronized with the estrous cycle.

Acetylcholine levels in the preoptic area of rat hypothalamus were found to change rhythmically, in synchronization with the estrous cycle. During the proestrous critical period, these levels demonstrated a drop followed by a sharp rise. The absence of this phenomena in males indicates sexual dimorphism. The close association between this newly detected hypothalamic acetylcholine rhythm and the estrous cycle suggests the possible involvement of acetylcholine in regulation of the estrous cycle.

Acetylcholine↗

Light, behavior, and biologic rhythms.

The most potent environmental synchronizer of "biologic clocks" in mammals is the light-dark cycle. The author describes experiments that identify a particular pathway--a derect connection between retina and hypothalamus--as the main route by which such biologic rhythms as the daily activity cycle, the estrous cycle, and the annual reproductive cycle are entrained to the external environment.

Animals↗

Comparison of synchronization of ovulation with timed insemination and exogenous progesterone as therapeutic strategies for ovarian cysts in lactating dairy cows.

The objective of this study was to compare the effectiveness of the Ovsynch and controlled internal drug releasing (CIDR) protocols under commercial conditions for the treatment of cystic ovarian disease in dairy cattle. A total of 401 lactating dairy cows with ovarian cysts were alternatively allocated to two treatment groups on the day of diagnosis. Cows in the Ovsynch group were treated with GnRH on Day 0, PGF2alpha on Day 7, GnRH on Day 9, with timed insemination 16-20 h later. Cows in the CIDR group were treated with a CIDR insert on Day 0 for 7 days; on Day 7, the CIDR was removed, and cows were treated with PGF2alpha. All cows in the CIDR group were observed for estrus and cows exhibiting estrus within 7 days following removal of the CIDR and PGF2alpha administration were inseminated. The outcomes of interest for this experiment were the likelihood to be inseminated, return to cyclicity (determined by a CL on Day 21), conception and pregnancy rates. Data for these variables were analyzed using logistic regression. The percentage of cows inseminated in the Ovsynch and CIDR groups were 82 and 44%, respectively. Cows in the Ovsynch group were 5.8 times more likely to be inseminated than cows in the CIDR group. Cows with a low BCS were 0.48 times less likely to be inseminated than cows with a high BCS. The percentage of cows with a CL on Day 21 for the Ovsynch and CIDR groups was 83 and 79%, respectively (P > 0.05). Cows with a low BCS were 0.49 times less likely to have CL on Day 21 than cows with a high BCS. Conception and pregnancy rates for cows in the Ovsynch group were 18.3 and 14.4%, respectively. Conception and pregnancy rates for cows in the CIDR group were 23.1 and 9.5%, respectively. There was no significant differences between conception or pregnancy rates in cows in both groups. Primiparous cows were 2.6 times more likely to conceive than multiparous cows. In conclusion, the results of this study suggested that fertility was not different between cows with ovarian cysts treated with either the Ovsynch or the CIDR protocols in this dairy herd. In addition, primiparous cows had an increased likelihood for conception compared to multiparous cows, and cows with a low BCS were less likely to be inseminated or have a CL on Day 21, regardless of treatment.

Animals↗

19-Hydroxylation of androgens by rat granulosa cells.

The transformation of androgens by rat granulosa cells was examined employing [19-C3H3]-, [1 beta-3H]-, and [1,2,6,7-3H]androgens as substrates. Rat granulosa cell homogenates incubated with [19-C3H3]androstenedione generated [3H] water and [3H]formic acid in a ratio of 8-9, indicating considerable 19-hydroxylation which was not followed by aromatization. This ratio remained relatively constant regardless of the time in the estrous cycle when the ovaries were removed, although there were large differences in the extent of the reactions. Parallel incubations with [1 beta-3H]]androstenedione showed that the aromatization of [19-C3H3]androstenedione in this tissue proceeds with a negative isotope effect of approximately 3, similar to that in human placenta. Incubation of the same substrates with granulosa cell cultures produced [3H]water and [3H]formic acid in ratios of 4-5 and showed a smaller negative isotope effect in the aromatization of [19-C3H3]androstenedione. FSH stimulation of the cell cultures had no influence on the ratio of 19-hydroxylation to aromatization with respect of either the duration of stimulation or the concentration of the pituitary hormone. Incubation of the cell cultures with [1,2,6,7-3H]androstenedione yielded tritium-labeled 19-hydroxy- and 19-oxoandrostendiones and estrogens in relative quantities corresponding to those expected from the [3H]water and [3H]formic acid formation. Virtually all of the products were found in the medium, with only trace quantities located intracellularly. Similarly, incubation of granulosa cell homogenates with [14C]androstenedione yielded [14C]19-oxygenated androgens in excess of [14C] estrogens. These results indicate that rat granulosa cells effect C-19-hydroxylation of androgens greater than that linked to aromatization and that the rat ovaries produce 19-oxygenated androgens in quantities exceeding those of estrogens. The excess 19-hydroxylation is synchronous with aromatization, but it is not known whether it is catalyzed by the same or a different enzyme. The formation of 19-oxygenated androgens in cell cultures indicates that they are distinct metabolites of androgens in the rat ovary and are not merely trapped transient aromatization intermediates.

1-Methyl-3-isobutylxanthine↗

Reproductive performance of lactating dairy cows treated with cloprostenol at the time of insemination.

The effect of intravenous cloprostenol treatment at the time of insemination on reproductive performance was consecutively evaluated in three different subpopulations of high producing lactating dairy cows: Study (1) early postpartum synchronized and fixed-time inseminated (about 50 days in milk) cows (n = 379: 187 control and 192 treated cows); Study (2) presumed high fertility cows first inseminated between 90 and 120 days postpartum (n = 248: 124 control and 124 treated cows); and Study (3) heat stressed repeat breeder cows (n = 183: 93 control and 90 treated cows). Data were analyzed using multiple regression methods. Study 1: Parity (primiparous versus multiparous), milk production, body condition score at AI, insemination season (cool versus warm period) and treatment were included in the analysis as potential factors affecting ovulation, double ovulation, return to estrus, and pregnancy to first AI and to second AI (first AI plus return AI) rates. Logistic regression analysis indicated that the final model for ovulation rate only included the interaction (P = 0.002) between insemination season and treatment. Cloprostenol treatment at insemination led to a 4.2-fold increase in the ovulation rate in cows inseminated during the warm period. There were no significant effects of treatment, parity, milk production, body score or the insemination season on the return to estrus rate. The only variables included in the final logistic model for double ovulation and pregnancy to first AI rates were treatment and season, respectively. Treatment led to a 2.6-fold increase (P = 0.001) in the double ovulation rate, whereas cows inseminated in the warm period were 2.1 times less likely (P = 0.007) to become pregnant at first AI compared to those inseminated in the cool season. The variables included in the final logistic model for the pregnancy rate to second AI were treatment and season. Cloprostenol given at AI increased the risk of pregnancy 1.9 times (P = 0.002), and cows inseminated during the warm season were two times less likely to become pregnant (P = 0.003). No significant interactions were found among these three dependent variables (double ovulation and pregnancy to first and to second AI rates). Study 2: Logistic regression analysis of all the dependent variables: return to estrus, and pregnancy to first and to second AI (first AI plus return to AI) rates indicated no significant effects of treatment, parity, days in milk, milk production or body score at AI. No significant interactions were found. Study 3: The final model for the pregnancy rate only included the interaction between parity (primiparous versus multiparous) and treatment. Days in milk, milk production and insemination number showed no significant effect on pregnancy rate. Cloprostenol treatment at insemination increased the pregnancy rate in primiparous repeat breeder cows (odds ratio: 3.6). The treatment group and parity showed significant (P < 0.0001) interaction. This interaction suggests that cloprostenol treatment of primiparous cows at insemination might enhance pregnancy yet have no effect in multiparous cows. Our findings indicate that cloprostenol administered at insemination promotes ovulation and double ovulation in lactating dairy cows. Cloprostenol treatment showed no benefit in cows with acceptable reproductive performance, suggesting that cloprostenol treatment at AI may only be useful in cows in which stress factors affect ovulation and in repeat breeder cows.

Animals↗

Ovarian atrial natriuretic peptide during the rat estrous cycle.

The changes in ovarian levels of immunoreactive atrial natriuretic peptide (irANP) and arginine vasopressin (irAVP) were observed during the estrous cycle of rat. We also demonstrated the synthesis of ovarian ANP. In adult 4-day cycling rats, ovarian level of irANP was found to be the highest on proestrus and was to be the lowest on diestrus. Ovarian irANP level inversely correlated with ovarian level of irAVP. On reverse-phase HPLC, two distinct peaks of ovarian irANP, high and low molecular weight forms, existed in the each stage of the estrous cycle. However, no significant changes in plasma and atrial concentrations of ANP were observed during the cycle. The rat ovary contained mRNA coding for ANP. These data showing the synchronized cyclic change of ovarian irANP and irAVP with the estrous cycle suggest that the ovary locally synthesizes ANP and ovarian ANP may play regulatory roles on the follicular fluid dynamics.

Analysis of Variance↗

Characterization of the periovulatory period in superovulated heifers.

Heifers (n=31) were superovulated with an FSH-P/cloprostenol regimen, and at 12 and 24 hours after the onset of estrus they were inseminated. Blood sampling for LH analyses and ultrasound scanning of the ovaries were performed at 4-hours intervals. The scanning, at which the first and last ovulations were recorded, was performed at 22.7 +/- 1.5 (mean +/- SD) and 31.0 +/- 1.5 hours after the LH peak, respectively. An average of 7.8 +/- 1.0 ovulations was monitored when the first ovulations were detected, while 2.8 +/- 0.7 ovulations occurred later. At 16 hours after detection of the first ovulations the oviducts were flushed and 5.6 +/- 0.5 fertilized and 2.3 +/- 0.3 unfertilized ova were isolated per animal. The fertilized ova displayed spherical pronuclei of synchronous development, and polyspermic penetration was not seen. At 24 hours after detection of the first ovulations the content of the remaining 3.3 +/- 0.5 nonovulatory follicles > 8 mm per animal was aspirated. Expanded cumulus investment was found in 69.4% of the oocytes, while 22.4% had abstricted the first polar body.

Journal Article↗

Prospects for improving reproductive performance through genetic selection.

Prospects for improving female fertility in dairy cattle using genetic selection are reviewed. Today's high producing cows have shorter estrous cycles, fewer standing events, shorter duration of estrus, and more frequent multiple ovulations. Although high milk production is often implicated as the cause of impaired fertility, the impact of inadequate body condition appears to be greater, as the latter has a significant impact on probability of conception, rate of embryonic loss, and proportion of anestrous animals. Genetic improvement of female fertility can be achieved by indirect selection for longevity or body condition score, or by direct selection for traits such as daughter pregnancy rate. Most leading dairy countries have implemented genetic evaluation systems for female fertility in the past decade, but refinement of these systems to account for hormonal synchronization, differences in the voluntary waiting period, exposure to natural service bulls, and other confounding factors is warranted. Recent work has focused on the development of data collection and genetic evaluation systems that will allow selection of bulls that have daughters that are resistant to common health disorders, including mastitis, lameness, ketosis, displaced abomasum, and metritis. Such systems will allow selection of animals that can remain healthy and fertile while producing large quantities of milk.

Animals↗

Effects of restricted food and water regimes on the estrous cycle of the rat.

The possibility that daily food and water presentation is a more potent synchronizer to the estrual rhythm of the female rat than the light-dark cycle was investigated by presenting food and water ad libitum only during the dark period or only during the light period of an LD 12:12. It was found that the daily feeding and drinking regimes did not synchronize the estrous cycle, but only disrupted the synchrony between the LD cycle and the estrous cycle. Because there was no significant difference between body weights of the groups, results cannot be interpreted in terms of under-nourishment effects on hormone synthesis, but rather in terms of the temporal characteristics of food and water intake.

Animals↗

Annual physiological changes in individually housed squirrel monkeys (Saimiri sciureus).

This study investigated whether annual changes in physiology occur in individually housed squirrel monkeys (Saimiri sciureus). Physiological measures were monitored for 20 months. Over the course of the study, all individually housed males and females exhibited clear annual changes in gonadal and adrenal hormone levels, and males exhibited species-typical changes in body weight. Females exhibited a typical pattern of hormonal changes, with elevations in gonadal steroids occurring during the same months as elevations in cortisol. Males, however, exhibited an atypical pattern, as elevations in hormone levels were not synchronized with each other; rather, elevations in testosterone occurred out of phase with changes in cortisol and body weight. The timing of annual events in individually housed subjects was compared to that in nearby social groups, in which the timing of the breeding season from year to year was determined by social group formations and was outside the naturally occurring breeding season. Elevations of ovarian and adrenocortical hormones in individually housed females were synchronized with indices of breeding in heterosexual social groups. Similarly, weight gain in males was associated with elevations in cortisol and, as with socially housed males, tended to precede seasonal breeding in the social groups. In contrast, annual testosterone elevations for individually housed males were not synchronized with breeding in nearby social groups. We conclude that direct physical interaction is not required for the annual expression of breeding readiness. Synchrony of seasonality among squirrel monkeys may be accomplished by distant social cues in females, but males may require physical interaction for complete synchrony of annual physiological changes.

Adrenal Cortex Hormones↗

Melatonin and photoperiodic time measurement: seasonal breeding in the sheep.

Well-established circadian physiology supports the view that photoperiodic time measurement utilizes the coincidence between the presence of light and a photosensitive phase of a 'biological clock' to alter reproductive status--the so-called external coincidence model of seasonal breeding. In this review, we examine the mechanism whereby photoperiod interacts with presumed suprachiasmatic nuclei activity to allow endogenous melatonin to normally synchronize reproductive activity to the optimal time of year. The Romney Marsh sheep is particularly explored as an experimental model. It is suggested that the on/off activity of seasonal reproduction may be a robust mechanism able to be predictably manipulated by the judicious use of the light/dark cycle and exogenous melatonin, but firmly based on circadian principles.

Animals↗

The expression of the clock protein PER2 in the limbic forebrain is modulated by the estrous cycle.

Daily behavioral and physiological rhythms are linked to circadian oscillations of clock genes in the brain and periphery that are synchronized by the master clock in the suprachiasmatic nucleus. In addition, there are a number of inputs that can influence circadian oscillations in clock gene expression in a tissue-specific manner. Here we identify an influence on the circadian oscillation of the clock protein PER2, endogenous changes in ovarian steroids, within two nuclei of the limbic forebrain: the oval nucleus of the bed nucleus of the stria terminalis and central nucleus of the amygdala. We show that the daily rhythm of PER2 expression within these nuclei but not in the suprachiasmatic nucleus, dentate gyrus, or basolateral amygdala is blunted in the metestrus and diestrus phases of the estrus cycle. The blunting of the PER2 rhythm at these phases of the cycle is abolished by ovariectomy and restored by phasic estrogen replacement suggesting that fluctuations in estrogen levels or their sequelae are necessary to produce these effects. The finding that fluctuations in ovarian hormones have area-specific effects on clock gene expression in the brain introduces a new level of organizational complexity in the control of circadian rhythms of behavior and physiology.

Animals↗

Embryo production and endocrine response in ewes superovulated with PMSG, with or without monoclonal anti-PMSG administered at different times.

Mature nonlactating Altamurana ewes (n = 168) were synchronized in the seasonal anestrus period with FGA-impregnated intravaginal pessaries for 12 d. In Experiment 1, 48 ewes were divided into a 3 x 4 factorial design for anti-PMSG monoclonal antibody (AP) bioassay test. Concomitant injections of PMSG (1000, 1500, 2000 IU) and AP (0, 1, 2, 3 microl/IU PMSG) were given, and ovarian response was evaluated by laparoscopy. In Experiment 2, 120 ewes were divided into 8 experimental groups (n = 15 per group). The ewes treated with 1000 or 1500 IU PMSG at -24 h from sponge removal were given AP intravenously at 50 h after pessary withdrawal, 12 or 24 h after the onset of estrus, while the controls did not receive AP. Blood samples were collected from ewes (n = 6) treated with 1500 IU PMSG with or without anti-PMSG. Ovarian response and embryo production were evaluated on Day 7 after sponge removal upon laparotomy. It was found that 1 microl AP was effective in neutralizing 1 IU PMSG. No significant differences in serum concentrations of progesterone were observed among the groups of superovulated ewes. Estradiol-17 beta levels were reduced following AP treatment 12 h after the onset of estrus. At a lower dosage of superovulatory treatment (1000 IU PMSG), AP injected at 12 or 24 h after the onset of estrus significantly lowered large follicles (P < 0.01) and increased the rate of ovulation (P < 0.05). Moreover, embryo production showed a more than two-fold increase (P < 0.01) of viable embryos following AP injection at 12 or 24 h after the onset of estrus (3.2 to 3.3 vs 1.3, with vs without anti-PMSG). It is concluded that superovulatory treatment with 1000 IU PMSG plus AP administered at a fixed time after the onset of estrus may improve ovarian response and the yield of viable embryos in ewes.

Journal Article↗

Control of ovulation with a GnRH analog in gilts and sows.

Methods for the control of ovulation with GnRH or the GnRH analog D-Phe6 -LHRH (GnRH-A), were evaluated in gilts and sows as the last step in development of a fixed-time Al protocol. This involved 3 field trials using 2,744 gilts (10 units) and 71,628 sows (33 units). In Trial 1, the GnRH-A (75 microg) was given subsequent to treatment with altrenogest for cycle control or eCG for the stimulation of uniform follicle development in gilts. The release of LH was followed by ovulations which commenced within 36.4 +/- 3.3 hr and were terminated at 39.0 +/- 2.8 hr after administration of GnRH-A. This degree of synchronization of ovulations enabled the use of fixed-time AI. Consequently, subsequent to pretreatment with altrenogest and eCG, in 10 production units 1,285 gilts received 50 microg GnRH-A and 1,459 gilts 500 IU hCG serving as positive controls (Trial 2); all the gilts were inseminated 24 and 42 hr after treatment. Pregnancy rate and piglet index (n of piglets per 100 first inseminations) following GnRH-A vs hCG were 78.8% and 779 vs 74.4% and 728, respectively (P < 0.05). In field trials with first litter gilts and multiparous sows (33 units holding from 250 to 6,000 sows), 1,000 IU eCG was used for estrus control after weaning and 25 microg or 50 microg GnRH-A were given 55 to 58 hours after eCG (n = 19,954 and 20,701) (Trial 3). Sows treated during the same time period with 300 microg GnRH plus 300 IU. hCG (n = 30,973) served as positive controls; all sows were inseminated 24 and 42 hours after treatment. Pregnancy rates for 50 microg GnRH-A, 25 microg GnRH-A and 300 microg GnRH plus 300 IU hCG were 83.0%, 81.7% and 80.7%, and the piglet indices 913, 899 and 880, respectively (P < 0.05). Unit size and parity had significant effects on fertility and productivity. In all studies, results with 50 microg GnRH-A were superior. In year-long studies, highest levels of fertility in response to these treatments were seen from December to May.

Journal Article↗

[Endocrine mechanisms of the synchronization of the estrous cycle in sheep].

Prostaglandin of the F2-alpha group (estrophan) led to a sharp decrease in serum progesterone concentration of 90-96% of Edibai female sheep eliminating the blocking hypothalamic center effect of the endogenous steroid hormone. The resultant conclusion was a possibility of artificial insemination of the above number of female sheep without finding out signs of wish in them. Their fertility from the first frontal insemination was not below 62-70%.

Animals↗