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Effect of stage of anestrus on the induction of estrus by the dopamine agonist cabergoline in dogs.

Beagle bitches were administered the dopamine D2 receptor agonist cabergoline in 3 groups of 5 animals each, starting on known days of the estrous cycle. Cabergoline treatment was started in either early anestrus (Days 93 to 108), mid-anestrus (Days 123 to 156), or late anestrus (Days 161 to 192) at doses of 5 ug/kg/d, per os, and was continued until the confirmation of induced proestrus or for 40 d. Reproductive parameters were compared with those in 5 control anestrous bitches (Days 90 to 150). In control bitches, the mean (+/- SEM) interval to the next proestrus (73+/-11 d) resulted in an interestrus interval (192+/-9 d) similar to that of the previous cycles (196+/-11 d). In 14 of the 15 cabergoline-treated bitches, the next proestrus occurred within 4 to 30 d, was premature in early and mid-anestrous bitches and developed with low variability within groups. The resulting intervals to proestrus in bitches treated with cabergoline in early anestrus (20+/-2 d), mid-anestrus (14+/-3 d) and late anestrus (6+/-1 d) resulted in interestrus intervals in those groups of 131+/-5, 166+/-7 and 196+/-2 d, respectively. In response to treatment, interestrus intervals were reduced (P<0.05) and more synchronous (P<0.05) in early and mid-anestrus bitches, and were more synchronous (P<0.05) in late-anestrous bitches compared with those of control bitches or those of the previous cycle. Periovulatory estradiol and progesterone profiles of induced cycles in treated bitches were similar to those of spontaneous cycles in control bitches. Four of 5 control bitches and 12 of the 14 responding cabergoline-treated bitches became pregnant and produced normal litters. Plasma prolactin concentrations at Days 2 and 5 of treatment (0.3+/-0.1 ng/mL) and at the onset of proestrus shortly before the end of treatment (0.4+/-0.1 ng/mL) were lower (P<0.05) than those present in anestrus prior to treatment (1.7+/-0.6 ng/mL) or in control bitches. Prolactin was also low at the onset of proestrus in control bitches (0.5+/-0.2 ng/mL). The results demonstrate that prolactin-lowering doses of the dopamine agonist cabergoline can terminate the normal obligate anestrus in dogs, and that the effect occurs more slowly in early anestrus than in mid or late anestrus.

Anestrus↗

The role of nutrition in the regulation of LH secretion during anestrus by the serotoninergic and dopaminergic systems in Mediterranean ewes treated with melatonin.

The steroid-dependent inhibition of LH secretion involves dopaminergic and serotoninergic systems but it is unclear how the plane of nutrition affects this inhibition during anestrus in melatonin treated ewes. Melatonin implants (18 mg) were inserted (Day 0) into ovariectomized, estradiol treated adult Rasa Aragonesa ewes on a high (H; n = 8) or low energy diet (L; n = 6) which were applied in early anestrus (Day 29-57) and late anestrus (Day 90-104). Cyproheptadine (0.1 mg/ kg), a serotoninergic (SHT2) receptor antagonist, was administered in early and late anestrus (Day 50 and 107) followed by pimozide (0.08 mg/kg), a dopaminergic2 receptor antagonist (Day 57 and 114). The H ewes had significantly higher LH concentrations (P < 0.05) before cyproheptadine treatment in early anestrus. The H and L ewes responded in a similar way to the antagonists in both early and late anestrus, except for L ewes who had a higher LH pulse amplitude after pimozide treatment in both periods (P < 0.05). During early anestrus, cyproheptadine tended to increase (P = 0.06) LH pulse frequency in L ewes and LH concentrations in H ewes. The LH secretion also increased in L ewes after pimozide administration during early anestrus (P < 0.05 for mean LH concentrations and LH pulse frequency and amplitude). However, pimozide dramatically increased LH secretion during late anestrus (Day 114) irrespective of the plane of nutrition (P = 0.06-0.08 for LH pulse frequency and P < 0.05 for LH concentrations and pulse amplitude). In melatonin treated Mediterranean ewes, the plane of nutrition appeared to modify the effect of dopaminergic and serotoninergic systems on the steroid-dependent inhibition of LH secretion throughout anestrus.

Anestrus↗

Physiological mechanisms controlling anestrus and infertility in postpartum beef cattle.

Postpartum infertility is caused by four factors: general infertility, lack of uterine involution, short estrous cycles and anestrus. The general infertility component is common to any estrous cycle and reduces potential fertility by 20 to 30%. Incomplete uterine involution prevents fertilization during the first 20 d after calving but is not related to anestrus. Short estrous cycles prevent fertility during the first 40 d after calving by causing the cow to return to estrus before pregnancy recognition occurs. Anestrus is the major component of postpartum infertility and is affected by several minor factors: season, breed, parity, dystocia, presence of a bull, uterine palpation and carryover effects from the previous pregnancy as well as two major factors: suckling and nutrition. These major factors have direct effects on anestrus but also interact with one or more other factors to control postpartum anestrus. Physiological mechanisms associated with anestrus involve blockage of the GnRH "pulse generator" in the hypothalamus, but other pathways also must be involved because bypassing the pulse generator is not an effective treatment for all cows. The primary cause of anestrus probably is different for different stages of anestrus. The mediating mechanisms for anestrus are not involved with prolactin, oxytocin, the adrenal or direct neural input from the mammary gland but are at least partially involved with blood glucose and the endogenous opioid peptide system. Management options to decrease the impact of anestrus and infertility include: 1) restrict breeding season to less than or equal to 45 d; 2) manage nutrition so body condition score is 5 to 7 before calving; 3) minimize effects of dystocia and stimulate estrous activity with a sterile bull and estrous synchronization; and 4) judicious use of complete, partial or short-term weaning.

Anestrus↗

Duration of the seasonal anestrus in sheep selected for fertility in a fall-lambing system.

Crossbred ewes (1/2 Dorset, 1/4 Rambouillet, and 1/4 Finnsheep) from a flock being selected for spring fertility, defined as ability to lamb following ram exposure in May and June in Virginia (37 degrees N latitude), were used to study the duration of the seasonal anestrus. In the first 3 yr of the study (1992, 1993, and 1995), mature ewes were divergently selected based on EBV for fertility, and the duration of anestrus was measured by continuously exposing the ewes to vasectomized rams equipped with marking harnesses from mid-January until approximately August 1. Only ewes that had lambed in the previous fall were used to ensure that ewes were in a comparable physiological state, and the same rams remained with the ewes in each year to avoid induction of estrus by introduction of novel rams. The duration of anestrus in high-fertility ewes (n = 26; mean fertility EBV of 12.6%) was 28.4 d, which was significantly less than the 70.2 d of anestrus observed for low-fertility ewes (n = 15; mean fertility EBV of .3%). Five high-fertility ewes did not exhibit a period of anestrus. The regression of number of days of anestrus on fertility EBV was -2.15 +/- .72 d/%. In yr 4 (1997), 11 high-fertility and two low-fertility ewes were evaluated. None of these ewes exhibited a clear seasonal anestrus; six unequivocally cycled continuously. Between January 23 and July 31, the mean duration of anestrus for these ewes was only approximately 11 d. The duration of anestrus for high-fertility ewes seems to be the shortest reported for temperate sheep breeds.

Anestrus↗

Descriptive epidemiology of anestrus in Michigan Holstein-Friesian cattle.

Twenty-two Michigan Holstein-Friesian herds were studied to determine the incidence and epidemiology of anestrus. In 3,309 lactations studied, 42% were classified as having exhibited preservice anestrus (no estrus detected by 70 d after calving). Organic reasons (pyometra, cystic follicles, static ovaries) were identified by palpation per rectum for 237 (23%) of the cows with preservice anestrus that were examined by a veterinarian. Postservice anestrus, defined as failure to show estrus within 35 d after an unsuccessful insemination, was identified in 790 (47%) of 1,691 lactations. Veterinary examination identified 104 (20%) of postservice anestrous cows as having this condition because of organic causes. The average cow with preservice anestrus had an increase of 30 d open compared to her herdmates, and the average cow with postservice anestrus had an increase of 37 d open. Anestrous cows produced more milk than their unaffected herdmates in both the current and the previous lactation. Analysis of composite lactation curves indicates that, for preservice anestrus, this additional milk production was obtained gradually over the entire lactation. Anestrous cows were culled at a significantly higher rate than their herdmates. Cows with preservice anestrus were more likely to have begun their lactation in the spring months.

Journal Article↗

A serial study of fluid balance during pregnancy, lactation and anestrus in goats.

Water and salt balance was studied in the same goats during pregnancy, lactation and anestrus. All goats increased their water intake during the course of pregnancy, but individual differences were large. In general, twin pregnant (TP) goats drank more water than single pregnant (SP) animals (3.4 +/- 0.4; N = 11 as compared to 2.5 +/- 0.3 litres/day; N = 5) during the last weeks of pregnancy. During lactation the high water intake TP goats persisted and the SP animals increased their intake to the same level as the TP goats. The water intake was reduced to about 2 1/day in both categories of animals during anestrus. Urine volume largely followed the changes in water intake in the individual animal. A continuous decrease in urine osmolality during the course of pregnancy occurred, but during lactation urine osmolality increased towards anestrus levels. TP goats generally retained more sodium than SP animals during pregnancy and during anestrus, whereas the figures were similar during lactation. Plasma Na, K and osmolality remained unchanged during pregnancy, lactation and anestrus, but a large fall in total plasma proteins and a moderate fall in hematocrit were observed during the course of pregnancy. Glomerular filtration rate of TP goats was elevated by about 35% during the 4th month of pregnancy, but did not differ from anestrus levels during the 3rd and 5th month or during lactation. Effective renal plasma flow was highest during the 3rd pregnancy month and then fell to reach lactation and anestrus levels during the 4th month of pregnancy. A few hours before parturition the animals became markedly dehydrated as shown by sudden increases in plasma Na, K, osmolality, total proteins and hematocrit. This water deficit was replenished within 26 h post-partum.

Anestrus↗

Nutritional anestrus in beef cows: concentrations of glucose and nonesterified fatty acids in plasma and insulin in serum.

Twenty-two nonlactating Hereford cows exhibiting normal estrous cycles were fed either maintenance (M) or restricted (R) diets until most of the R cows became anestrus; R cows then were fed 160% of the M diet until estrous cycles resumed. Concentrations of progesterone, glucose, insulin and nonesterified fatty acids (NEFA) were determined in weekly blood samples. Blood also was collected frequently, before and after i.v. infusion of 300 ml of a 40% glucose solution, to evaluate responses in blood concentrations of glucose and insulin when cows were exhibiting normal estrous cycles, when R cows were initiating anestrus, during anestrus, and at the reinitiation of estrous cycles. Losses in BW and body condition score in R cows were associated with reduced (P less than .01) concentrations of glucose and insulin and greater (P less than .01) concentrations of NEFA in blood plasma compared with those of M cows. During normal estrous cycles, disappearance of infused glucose from plasma and concentrations of insulin in serum were similar for R and M cows. Glucose disappearance from plasma was retarded and serum concentrations of insulin remained increased for a longer time after glucose infusion in R at the start of anestrus compared with M cows (P less than .01). Similarly, during anestrus, the rate of glucose disappearance was slower for R cows (P less than .01). During refeeding of R cows, disappearance of infused glucose was similar for R and M cows. In conclusion, reduced concentrations of glucose and insulin and increased concentrations of NEFA in blood were associated with nutritional anestrus and the glucoregulatory effects of insulin were compromised during nutritional anestrus.

Anestrus↗

Body condition and suckling as factors influencing the duration of postpartum anestrus in cattle: a review.

Prolonged postpartum anestrus is a main factor limiting reproductive efficiency in cattle, particularly in Bos indicus and Bos taurus/Bos indicus cows from tropical regions, because it prevents achievement of a 12 month calving interval. During anestrus, ovulation does not occur despite ovarian follicular development, because growing follicles do not mature. Although many factors affect postpartum anestrus, nutrition and suckling are the major factors influencing the resumption of postpartum ovarian cycles, as they affect hypothalamic, pituitary and ovarian activity and thus inhibit follicular development. Under-nutrition contributes to prolonged postpartum anestrus, particularly among cows dependent upon forages to meet their feed requirements and it apparently interacts with genetic, environmental or management factors to influence the duration of anestrus. The nutritional status or balance of an animal is evaluated through body condition score (BCS), as it reflects the body energy reserves available for metabolism, growth, lactation and activity. There is a converse relationship between energy balance and time to resumption of postpartum ovarian activity; inadequate nutrient intake results in loss of weight and BCS and finally cessation of estrous cycles. Suckling interferes with hypothalamic release of GnRH, provoking a marked suppression in pulsatile LH release, resulting in extended postpartum anestrus. The effects of suckling on regulation of tonic LH release are determined by the ability of the cow to identify a calf as her own or as unrelated. Vision and olfaction play critical roles in the development of the maternal-offspring bond, allowing the cow to identify her own calf, and abolition of both senses attenuates the negative effects of suckling on LH secretion. Thus, the maternal-offspring bond is essential for prolonged postpartum suckling-induced anovulation, and the suppressive influence of suckling is independent of neurosensory pathways within the teat or udder.

Anestrus↗

The postpartum buffalo. II. Acyclicity and anestrus.

Prolonged postpartum acyclicity (absence of ovarian cyclic activity) and anestrum (absence of overt estrous signs) are major sources of economic loss to buffalo breeders. Studies on the epidemiology of these two problems are highly recommended to achieve successful control. Review of the available literature on controlled studies in dairy buffaloes revealed that first ovulation as detected by rectal palpation and progesterone analysis occurred between 28-71 and 24-55 days, respectively, after calving. Postpartum estrus in the same studies occurred between 44 and 87 days. Reports concerned with data compiled from breeding records of research stations, breeding farms and small holders where estrus is a subjective measure, gave much longer periods. Also data from Egypt, India and Pakistan indicate that only 34-49% of buffaloes showed estrus during the first 90 days after calving and 31-42% remained anestrus for more than 150 days. In swamp buffaloes both postpartum ovulation and estrus are more delayed than in dairy buffaloes. The role of suckling, nutrition, body condition score at calving, milk yield, parity, season of calving and other minor factors were discussed. First postpartum ovulation is frequently followed by one or more short estrous cycles (<18 days). Long anovulatory and anestrous periods due to prolonged inter-luteal phase were reported to occur after short cycles. Also long anestrous periods due to cessation of cyclic activity (true anestrus) for 3 or more weeks and prolonged luteal activity for 28 days or more were described to occur in about 25 and 8-11% of the buffaloes, respectively, after the first or second ovulation. These cycle irregularities certainly impose difficulties on estrus detection programs in postpartum buffaloes. Four main forms of anestrus i.e. true anestrus (inactive ovaries and small and medium sized anovulatory follicles), subestrus, prolonged luteal activity and ovarian cysts in addition to pregnancy are reviewed in this article. Differentiation between true anestrus and subestrus is particularly important in buffaloes because of their weak estrous signs. However, the accuracy of a single rectal palpation of the ovaries is limited with an overestimation of the frequency of true anestrus due to misdiagnosis of the corpus luteum. The possible causes are discussed.

Anestrus↗

Concurrent pulsatile secretion of luteinizing hormone and follicle-stimulating hormone during different phases of the estrous cycle and anestrus in beagle bitches.

The 6-h secretory profiles of LH and FSH and the possible concordance between the episodic release of LH and FSH were studied in 6 beagle bitches during early, mid-, and late anestrus and during the follicular and luteal phases of the estrous cycle. Plasma samples were obtained at 10-min intervals via jugular venipuncture. In all stages of anestrus and in the luteal phase, FSH and LH secretion was pulsatile. All FSH pulses coincided with LH pulses. However, the mean duration of the FSH pulse (115 min) was significantly longer than that of the LH pulse (72 min). The basal plasma LH concentration was low compared with the maximum peak levels, whereas FSH pulses were characterized by relatively low peaks compared with the basal levels. In contrast to the basal plasma LH levels and the area under the curve (AUC) for LH, the basal plasma FSH levels and the AUC for FSH increased significantly as anestrus progressed. During the follicular phase, the secretory pattern of LH was characterized by frequent increases of short duration. During this phase, the basal plasma FSH concentration was relatively low, whereas the basal plasma LH level was high in comparison with that in the other phases of the estrous cycle. The luteal phase was characterized by an increased frequency of LH pulses, a shorter duration of the LH peaks, and a tendency to a lower amplitude of both LH and FSH peaks compared with values observed during anestrus. It is concluded that in the bitch, FSH and LH pulses are released in concordance and that progression from early to late anestrus is associated with an increase in basal plasma FSH concentration without a concomitant rise in basal plasma LH concentrations. The latter suggests that in the bitch an increase in circulating FSH should be considered to be a critical event required for the initiation of ovarian folliculogenesis and consequently for the termination of anestrus.

Anestrus↗

An essential role for ovarian inhibin in pineal gland-mediated anestrus in Syrian hamsters.

When transferred from long photoperiod (LP) to short photoperiod (SP), female Syrian hamsters exhibit depressions of follicle stimulating hormone (FSH) and follicular development, cessation of ovulation, and marked ovarian interstitial tissue hyperplasia. Pinealectomy prevents these effects of SP. The object of this study was to determine the role of inhibin in the regulation of FSH during SP-induced anestrus. Adult LSH/SsLak hamsters maintained in LP (LD 14:10 hr) were transferred to SP (LD 8:16 hr) on the day of estrus, and groups of animals killed at either 16.00 hr on proestrus or 08.00 hr on estrus during each of five consecutive 4-day estrus cycles after transfer. Groups of females that became anestrus in SP were killed either at 08.00 or 16.00 hr, 12 days after the last observed estrus discharge. Compared to LP controls, serum FSH levels on estrus increased significantly (P<0.01) during the first two cycles in SP before declining to concentrations that were significantly lower than control values (P<0.01). Serum inhibin levels increased significantly by the third, fourth, and five days of estrus in SP. Regression analysis revealed a significant inverse correlation between serum inhibin and FSH levels on estrus during SP exposure (P=0.021) but not on proestrus. Relative levels of inhibin alpha- and betaA-subunit mRNAs were lower in ovaries from SP proestrus and anestrus females killed at 16.00 hr as compared to those from proestrus LP controls; they were elevated in ovaries from SP estrus and anestrus females killed at 08.00 hr compared to those from estrus LP controls. The absence of antral follicles on estrus in the last cycles of SP and anestrus suggests that the increase in circulating inhibin and inhibin mRNAs may be derived from hyperplastic interstitium. These observations suggest that inhibin may play an essential role in suppressing FSH secretion during pineal gland-mediated anestrus in Syrian hamsters.

Anestrus↗

Effects of varying energy intake and sire breed on duration of postpartum anestrus, insulin like growth factor-1, and growth hormone in mature crossbred cows.

Objectives of this study were to evaluate effects of seven sire breed groups and three levels of daily ME intake (DMEI = 132 or 189 kcal ME/kg BW(0.75) or ad libitum), beginning 5 mo prepartum, on BCS, length of postpartum anestrus, and circulating concentrations of IGF-1 and GH in F1 cows (six to eight cows per sire breed in each DMEI group) out of Angus or Hereford dams. At the initiation of the study, BW were 522, 530, 548, 572, 575, 577, and 595 kg for cows sired by Longhorn, Galloway, 1960s Hereford or Angus, 1980s Hereford or Angus, or Nellore, Salers, and Shorthorn bulls, respectively (SE = 13; P < 0.001 for sire breed). After 4 mo on DMEI treatment during the pre-partum period, cows fed 132 kcal of ME/kg BW(0.75)gained little to no BW; cows fed 189 kcal ME/kg BW(0.75) gained 50 kg; and cows fed ad libitum gained 70 kg (all groups differ P < 0.05). Concentrations of progesterone in weekly blood samples collected 2 to 14 wk after calving were used to establish when normal luteal function resumed to predict length of postpartum anestrus. Length of anestrus was affected by level of DMEI in cows sired by Galloway, Longhorn, and Nellore bulls, but not other breeds (P < 0.02 for interaction of sire breed and DMEI). Level of DMEI, but not sire breed, affected (P < 0.01) BCS at wk 2 postpartum. Concentrations of IGF-1 at wk 2 postpartum differed (P < 0.001) due to sire breed, and changes in concentrations of IGF-1 from wk 2 to 14 were influenced (P < 0.03) by the interaction of sire breed and level of DMEI; which was primarily the result of differences in rate of decrease over time among different sire breed x level of DMEI groupings. Concentrations of GH did not differ due to sire breed but varied (P < 0.001) due to the interaction of DMEI and week postpartum, for which concentrations of GH did not differ at wk 2 but increased over time at rates that were inversely proportional to level of DMEI. Length of anestrus was negatively associated (P < 0.05) with day of calving, BCS, and BW. When effects of sire breed and level of DMEI were accounted for (residual correlation), length of anestrus was inversely associated (P < 0.01) with IGF-1 concentrations. Breed of sire influenced length of postpartum anestrus and energy balance, as predicted by IGF-1, in crossbred cows fed varying levels of DMEI.

Anestrus↗

Absorption and systemic availability of two synthetic growth hormone secretogogues and transport of glucose by the proximal small intestine of anestrus dogs after administering estradiol.

Pharmacokinetics for one growth hormone secretogogue (NNC 26-0722), but not for another (NN703), differ between dogs in estrus or anestrus. We examined if the differences could be mimicked by administering estradiol during anestrus and if there was a relationship with rates of small intestine absorption. Pharmacokinetics for oral doses of NN703 (1.0-1.6 mg kg(-1)) did not differ among dogs in estrus, anestrus, or anestrus and given estradiol for 1 week (days 1, 3, and 6; 40 micro g kg(-1)), whereas plasma concentrations of NNC 26-0722 increased from undetectable in untreated, anestrus dogs to several hundred nanograms per milliliter in dogs given estradiol, with maximal concentrations measured 5 min after oral dosage. Estradiol treatment increased small intestinal absorption of NNC 26-0722 by 100% (P<0.05), but did not increase absorption of NN703, and caused a 64% increase in carrier-mediated glucose transport at 50 mmol l(-1) (P<0.05) due to increased densities of transporters. These findings indicate estrus and estradiol enhance absorptive functions of the dog proximal small intestine and can affect pharmacokinetics for some orally administered drugs.

Administration, Oral↗

Resynchronization of ovulation and timed insemination in lactating dairy cows, II: assigning protocols according to stages of the estrous cycle, or presence of ovarian cysts or anestrus.

Pregnancy rates were compared in lactating dairy cows (n = 1083) assigned to protocols for resynchronization of ovulation based on stages of the estrous cycle, or presence of ovarian cysts or anestrus. Cows were detected not pregnant by ultrasonography 30 d after a previous AI (study day 0) and classified as diestrus, metestrus, proestrus, with ovarian cysts or anestrus. Cows in diestrus (January-May) were assigned to either Ovsynch (GnRH day 0, PGF2alpha day 7, GnRH day 9, and timed-AI [TAI] 16 h later; n = 96), or Quicksynch (PGF2alpha day 0, estradiol cypionate [ECP] day 1, AI at detected estrus [AIDE] on day 2, or TAI on day 3; n = 96). Cows in diestrus (June-December) were assigned to either Ovsynch (n = 156) or Modified Quicksynch (PGF2alpha day 0, ECP day 1, AIDE days 2 and 3, and to Ovsynch on day 4 if not detected in estrus; n = 142). Cows in metestrus were assigned either to Ovsynch (n = 68), Heatsynch (GnRH day 0, PGF2alpha day 7, ECP day 8, AIDE day 9, or TAI day 10; n = 62), or GnRH + Ovsynch (GnRH on day 0, followed by Ovsynch on day 8; n = 64). Cows in proestrus, with ovarian cysts, or anestrus were assigned to either Ovsynch (proestrus n = 89, ovarian cysts n = 97, anestrus n = 8) or GnRH + Ovsynch (proestrus n = 87, ovarian cysts n = 109, anestrus n = 9). Pregnancy rate was evaluated 30, 55 and 90 d after resynchronized AI. For cows in diestrus (January-May), pregnancy rates were higher for Ovsynch (35.9, 29.2 and 26.0%) than for Quicksynch (21.7, 16.7 and 15.6%). For cows in diestrus (June-December), pregnancy rates were similar for Ovsynch (34.4, 24.0 and 23.6%) and Modified Quicksynch (27.1, 26.2 and 21.6%). For cows in metestrus, pregnancy rates were higher for GnRH + Ovsynch (33.3, 24.5 and 20.3%) than for Heatsynch (20.3, 12.9 and 9.8%). For cows with ovarian cysts, pregnancy rates were higher for GnRH + Ovsynch (30.3, 26.6 and 22.9%) than for Ovsynch (20.2, 18.5 and 14.7%). Assignment to resynchronization protocols based on the stages of the estrous cycle, or presence of ovarian cysts improved pregnancy rates.

Anestrus↗

Effects of follicle stimulating hormone (FSH) on follicular development, oocyte retrieval, and in vitro fertilization (IVF) in ewes during breeding season and seasonal anestrus.

Administration of FSH increases the number of developing follicles, and affects oocyte health and cleavage rate. To determine the optimal level of FSH treatment, studies were conducted during the normal breeding season and seasonal anestrus. In Experiment 1, ewes were implanted with SyncroMate-B (SMB; norgestomet) for 14 days during the breeding season. Beginning on day 12 or 13 after SMB implantation, ewes were treated with saline (control; n=10), or treated with FSH for two days (2D; n=9) or three days (3D; n=10). In Experiment 2, conducted during seasonal anestrus, ewes were implanted with SMB for 14 days (n=23) or were not implanted (n=26). The SMB-implanted and nonimplanted ewes were assigned to one of three treatments as in Experiment 1: control (n=13), 2D (n=21) or 3D (n=15). In Experiments 1 and 2, ewes were laparotomized to count the number of follicles < or = 3 mm and > 3 mm and to retrieve oocytes. Healthy oocytes from each treatment were used for IVF. In Experiment 3, ewes (n=6) were implanted twice with SMB for 14 days during seasonal anestrus. Ewes were injected with FSH for 2 days, and the oocytes were collected and fertilized as in Experiments 1 and 2. In Experiment 1, FSH-treatment increased (P < 0.05) the number of follicles > 3 mm, the number of oocytes retrieved from follicles < or = 3 mm and > 3 mm, the proportion of healthy oocytes, and the number of oocytes used for IVF. Oocytes from control and 2D ewes had greater (P < 0.01) cleavage rates than 3D ewes (68% and 71% vs. 42%). In Experiment 2, implanted and nonimplanted ewes had similar (P > 0.05) numbers of follicles, total oocytes, and healthy oocytes; therefore, data were combined. The FSH treatment increased (P < 0.01) the number of follicles > 3 mm, and the number of oocytes recovered from follicles > 3 mm. The recovery rate of oocytes and the percentage of healthy oocytes were similar for control and FSH-treated ewes. The cleavage rate in Experiment 2 ranged from 4 to 16%. In Experiment 3, the cleavage rate for ewes treated twice with SMB was 27% which tended to be greater (P < 0.07) than for the 2D ewes that received one SMB implant in Experiment 2. These data indicate that FSH increased the number of developing follicles and the number of healthy oocytes retrieved from ewes during the breeding season and seasonal anestrus. However, cleavage rates during seasonal anestrus were lower than during the normal breeding season in both FSH-treated and control ewes. Treatment of ewes for 2 days with FSH resulted in a greater cleavage rate than treatment of ewes for 3 days.

Anestrus↗

Increased LH pulse frequency and estrogen secretion associated with termination of anestrus followed by enhancement of uterine estrogen receptor gene expression in the beagle bitch.

The relationships among pulsatile LH secretion pattern, estrogen secretion, and expression of the uterine estrogen receptor gene were examined throughout the estrous cycle in beagle bitches. In Experiment 1, blood samples were collected from 30 bitches every 10 min for 8 h from a cephalic vein during different phases of the estrous cycle. An increase in the mean plasma levels of LH occurred from mid to late anestrus (P < 0.01). The LH pulse frequency increased (P < 0.01) from late anestrus to proestrus, and was strongly correlated (r = 0.96, P < 0.001) with the mean plasma level of estradiol-17 beta (E2). In Experiment 2, middle uterine samples, including the myometrium and endometrium, from 18 bitches were taken at 6 stages of the estrous cycle. The total number of estrogen receptors and nuclear estrogen receptor and its mRNA levels in the uterus also increased (P < 0.01) from late anestrus to proestrus. Mean plasma E2 level and the number of uterine estrogen receptor were positively correlated (r = 0.81, P < 0.05). In Experiment 3, nine bitches were ovariectomized in mid anestrus. Two weeks later they received a single injection of 10 or 50 micrograms/kg, i.m., estradiol benzoate. The number of uterine estrogen receptor and their mRNA levels for ovariectomized bitches were low, but increased (P < 0.05) after treatment with a low dose of estradiol benzoate. These results suggest that increases in LH pulse frequency and estrogen secretion are associated with termination of anestrus and that subsequent enhancement of uterine estrogen receptor expression may be up-regulated by estradiol.

Anestrus↗

Concentrations of reproductive hormones in canine serum throughout late anestrus, proestrus and estrus.

Concentrations of estradiol, progesterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin in serum from 6 bitches bled daily for at least 45 days before the onset of proestrus, during proestrus and estrus were determined by radioimmunoassay. Mean concentrations of estradiol in serum were high early in the sampling period (20 to 46 pg/ml) and appeared to decrease prior to the onset of proestrus (8 to 19 pg/ml). There were sporadic increases in serum concentrations of LH throughout the sampling period in each bitch. Five of the 6 bitches sampled had increased serum concentrations of LH following the low mean concentration of estradiol just before the onset of proestrus. Mean concentrations of FSH were highest during anestrus (240 to 294 ng/ml) and near the time of the preovulatory surge of LH (297 ng/ml) and were lowest during proestrus (131 to 200 ng/ml). The mean concentration of progesterone for the 6 bitches remained at less than 1.0 ng/ml throughout late anestrus, but increased to greater than 1.0 ng/ml the day of the maximum mean concentration of LH (preovulatory LH surge). Mean concentrations of prolactin were variable throughout the sampling period and demonstrated no consistent pattern among bitches. The results of the current investigation suggest that neither the canine ovary nor pituitary are quiescent during anestrus. The bitch appears to have sufficient FSH present during anestrus for follicular growth. Serum concentrations of LH appear to increase prior to the onset of proestrus when concentrations of estradiol are lowest, possibly inducing a new follicular phase. Progesterone and prolactin do not appear to be involved in the termination of anestrus in the bitch.

Anestrus↗

Time-course of thyroid hormone involvement in the development of anestrus in the ewe.

It is well established that the thyroid gland is essential for termination of seasonal reproductive activity in a variety of birds and mammals. In the present study, we examined when during the breeding season the thyroid exerts this effect in female sheep. Previous results suggest that the presence of thyroid hormones during the first 4-6 wk (20-25%) of the breeding season is not sufficient for the neuroendocrine changes that lead to anestrus. We therefore hypothesized that thyroid hormone action is exerted at some point during the latter 75-80% of the breeding season. To test this hypothesis, ewes thyroidectomized early in the breeding season received replacement of thyroxine at various times to create gaps during the mid- to late breeding season when thyroid hormones were absent. We then examined the effect, if any, of this absence on development of seasonal neuroendocrine anestrus. Each ewe was ovariectomized and treated with a constant-release Silastic capsule containing estradiol. Serum concentrations of LH were used as an index of seasonal changes in reproductive neuroendocrine activity. We found that when thyroid hormones were removed for a 60-day period in mid- to late breeding season (from mid-Oct. to late Dec., which is approximately 40% of the entire breeding season), anestrus still developed at the normal time. We conclude, therefore, that thyroid hormones need not be present for much of the breeding season (mid-Sept. through late Dec.) for anestrus to develop in the ewe. Rather, we postulate that thyroid hormones need to be present for only a brief period of time near the end of the breeding season for the neuroendocrine changes that lead to anestrus.

Anestrus↗