The effect of bpV(HOpic) on in vitro activation of primordial follicles in cultured swine ovarian cortical strips

The vanadate‐derivative dipotassium bisperoxo (5‐hydroxy‐pyridine‐2‐carboxylic) oxovanadate (V) (bpV(HOpic)), a pharmacological inhibitor of phosphatase and tensin homolog (PTEN), has been used in ovarian follicle culture systems for activation of follicular growth in vitro and suggested to be responsible for primordial follicle survival through indirect Akt activation. For pig ovarian tissue, it is still not clear which culture medium needs to be used, as well as which factors and hormones could influence follicular development; this also applies to bpV(HOpic) exposure. Therefore, ovarian cortical strips from pigs were cultured in 1 µM bpV(HOpic) (N = 24) or control medium (N = 24) for 48 hr. Media were then replaced with control medium and all tissue pieces incubated for additional 4 days. The strips were embedded in paraffin for histological determination of follicle proportions at the end of the culture period and compared to histological sections from tissue pieces without cultivation, which had been embedded right after preparation; comparison of healthy follicles for each developmental stage was performed to quantify follicle survival and activation. After 6‐day culture, follicle activation occurred in tissue samples from both cultured groups but significantly more follicles showed progression of follicular development in the presence of 1 µM bpV(HOpic). The amount of non‐vital follicles was not significantly increased during cultivation. BpV(HOpic) affects pig ovarian follicle development by promoting the initiation of follicle growth and development, similar as in rodent species and humans.     

Effects of Calf Removal at Parturition on Postpartum Ovarian Activity in Zebu (Bos indicus) Cows in the Humid Tropics

To assess endocrine and morphological responses of ovaries to total weaning at parturition, 6 Zebu (Bos indicus) cows 5 years or older were investigated. Following parturition, blood samples were collected daily during the first month and twice weekly thereafter until day 60 to determine concentrations of progesterone (P₄) and prostaglandin F_2α metabolite.It took between 25 to 32 days to complete uterine involution.The prostaglandin metabolite remained elevated for a mean period of 14.2 days (range, 4-21) postpartum. Five of the animals resumed cyclicity with a short estrous cycle starting between days 7 to 34 and lasting between 7 and 14 days. No estrous behavior was recorded prior to the short estrous cycles, but subsequent normal-length estrous cycles were all preceded by signs of estrus. In the 1 animal that resumed cyclicity with an estrous cycle of normal length on day 37 (length 20 days), the cycle was preceded by estrous behavior.Progesterone concentrations reached a mean maximum of 4.8 nmol liter⁻¹ during the short estrous cycles, and prostaglandin metabolite concentrations peaked while P₄ concentrations were decreasing. P₄ concentrations reached a mean maximum of 12.2 nmol liter⁻¹ during the estrous cycles of normal length. The interval from parturition to the first estrous cycle of normal length varied between 16 and 48 days, and the length of the cycle was 18 to 22 days.Starting 2 days postpartum, ovaries from 5 of the cows were scanned by ultrasonography every second day until day 30 postpartum. Medium-sized follicles were detected between days 4 to 7 postpartum in 4 of the scanned cows that later had short estrous cycles. The time between parturition and the appearance of the first dominant follicle was 7.6 days (range 6-10 days). The interval between parturition and the appearance of the first ovulatory-sized follicle was 10.2 days (range 8-13 days). In 3 of the scanned cows this ovulatory-sized follicle ovulated.We conclude that cyclic ovarian activity in Zebu cows can start early in the postpartum period in the absence of offspring, and that short luteal phases, not preceded by estrous behavior, may play an important role in establishing normal postpartum ovarian activity.     

β-defensins gene expression in tissues of the crossbred and Tibetan pigs

Delayed resumption of postpartum oestrous cycles in dairy cows is linked to low plasma insulin status and associated changes in ovarian function. Previous studies showed that plasma insulin can be raised by increasing dietary starch concentration, but it is not known if rumen digestible starch and bypass starch would be equally effective. The objective of the two experiments reported here was to investigate whether site of dietary starch digestion influences metabolic hormones and ovarian function. Cows were fed on a standard diet from calving until 40 days postpartum and then fed on treatment diets until 70 days postpartum. In Experiment 1, six cows were transferred to each of five diets with a starch content of 190 g/kg DM, of which 0.40, 0.45, 0.50, 0.55 or 0.60 was bypass starch. In Experiment 2, eight cows were transferred to each of four diets containing concentrates with low (L) or high (H) starch content and predominantly grass (G) or maize (M) silages in a factorial design. Between 60 and 70 days postpartum, plasma insulin was not influenced by site of starch digestion in Experiment 1, but insulin was increased by H (P = 0.020) and M (P = 0.048) in Experiment 2. Dietary bypass starch content negatively influenced plasma glucagon concentration (P = 0.046) and the insulin to glucagon ratio (P = 0.005) in Experiment 1. Growth hormone, insulin-like growth factor-I and leptin did not vary among diets in either experiment. The number of small (< 5 mm) ovarian follicles before ovulation tended (P = 0.056) to decrease with increasing dietary bypass starch content and was negatively related (P = 0.046) to plasma glucagon in Experiment 1, but there was no other dietary effect on ovarian function in either experiment. The main conclusion from these experiments is that rumen digestible starch and rumen bypass starch can be equally effective for maintaining plasma insulin and ovarian function of high-yielding dairy cows in early lactation.     

Follicular turnover and hormonal association in postpartum goats during early and late lactation

To clarify the effect of lactation period on ovarian follicular activity and associated hormonal levels in goats, six goats were monitored daily by ultrasonographic examination with blood sampling during early (Days 5 to 25; Day 0 was the day of kidding) and late (Days 40 to 60) lactation. While the presence of a corpus luteum of pregnancy retarded follicular growth in the ipsilateral ovary until Days 11-13 postpartum, the total follicular number (TFN) and area (TFA) increased during late lactation due to the significant increase in the number of medium- and large-sized follicles and decrease in the number of small follicles. Four goats showed a similar pattern of follicular development during the period studied characterized by the emergence of five and six waves during the early and late lactation, respectively. The largest follicle diameter of the first three waves monitored during early lactation was significantly smaller as compared with the diameter of those existing during late lactation. TFN showed a positive correlation with FSH but showed a negative correlation with immunoreactive (ir-) inhibin and estradiol during the postpartum period. TFA was positively correlated with ir-inhibin, estradiol and PRL and negatively correlated with FSH during the monitored periods. The plasma levels of ir-inhibin and progesterone were significantly higher during late lactation compared with the levels recorded during early lactation. Ir-inhibin levels showed a significant positive correlation with LH and estradiol during early and late lactation but showed a negative correlation with FSH during the whole lactation period. LH was positively correlated with estradiol and PRL during early and late lactation, respectively. These results suggest that the lactation period has a detrimental effect on ovarian activity during the early postpartum period in goats.     

Factors that affect ovarian follicular dynamics in cattle.

Studies of ovarian follicular dynamics in cattle may lead to methods for improving fertility, for synchronizing estrus with more precision, and for enhancing superovulatory responses. Within an estrous cycle, two or three large (> 10 mm) follicles develop during consecutive waves of follicular growth. The last wave provides the ovulatory follicle, whereas preceding wave(s) provide follicles that undergo atresia. The life span of large follicles seems to depend on the pulsatile secretion of LH; decreased frequency of LH pulses results in atresia of large follicles. Aromatase activity in the walls of the largest follicles is greatest during the first 8 d of the estrous cycle and decreases by d 12. Steroidogenesis of the largest and second-largest ovarian follicles differs on d 5, 8, and 12 of the estrous cycle. Follicular dynamics are altered by negative energy balance and lactation. The number of large follicles and concentration of estradiol during the preovulatory period differs between postpartum lactating and nonlactating cows. Dietary fats stimulate follicular growth when they are fed to increase energy balance. Administration of bovine somatotropin decreases energy balance and has a differential effect on ovarian follicular responses; growth of the largest follicle does not change, but growth of the second-largest follicle is stimulated by somatotropin. Studies of follicular dynamics in lactating cows demonstrate changes in ovarian function associated with energy balance that may be related to inefficient reproductive performance of cows producing high yields of milk.     

Associations of prepartum body condition score with occurrence of clinical endometritis and resumption of postpartum ovarian activity in dairy cattle

This study was performed to investigate the effect of periparturient body condition score on the occurrence of clinical endometritis and postpartum resumption of ovarian activity in dairy cows. Eighty-seven lactating Holstein cows, fed with a total mixed ration diet, were included into the study. Body condition scoring (using a 5-point scale with quarter-point divisions) was performed by the same investigator using the visual technique every 2 weeks, from 2 weeks before until 6 weeks after calving. Palpation of the reproductive tract and ultrasonographic assessment of ovaries for detection of corpus luteum using a rectal linear probe was also performed at 2, 4, and 6 weeks after calving. Cows with clinical endometritis had significantly lower body condition score (BCS) than normal cows at all weeks pre- and postcalving, and cows that did not ovulate until 45 days after calving had a significantly lower BCS pre- and postpartum. Cows that did not ovulate until 45 days after calving also lost more BCS from 2 weeks before to 4 weeks after calving. Besides, first ovulation after calving take occurred later in cows with clinical endometritis compared to normal cows (P?<?0.05). In conclusion, low BCS is a risk factor for postpartum endometritis and delayed cyclicity in dairy cows. BCS loss from dry-off to early lactation and occurrence of clinical endometritis can significantly affect postpartum ovarian activity.     

Turnover of Dominant Follicles prior to First Ovulation and Subsequent Fertility in Postpartum Dairy Cows

Contents: The ovaries of 64 multiparous Holstein cows were scanned twice weekly by ultrasonographyfrom Day 6 postpartum until 60 days or return to normal estrous cycle, and turnover of dominant follicles in relation to subsequent fertility were determined. The mean (SD) number of days from parturition to detection of the first postpartum dominant follicle (10 mm) was 9.0 0.5, which was predominantly located in the ovary contralateral to previous pregnancy in 54/64 cows. The first postpartum dominant follicle ovulated in 34 cows, failed to ovulate in 18, became cystic in 10, and only 2 were diagnosed as ovarian quiescent which delayed the detection of dominant follicles until days 25 and 30 postpartum. Number of dominant follicles in cows with anovulated first dominant follicle was 2.8. 1.1 prior to first ovulation (range 2–5). Growth and regression of cystic follicles were observed 24 times in 6110 cows, and a total of 20 cystic follicles persisted 32.7 9.0 days (range 18–58 days) except one which persisted for 124 days. Conception rate was 89.5% and postpartum interval to conception within 180 d after calving was 96.7 days in cows detected with less than 3 dominant follicles before first ovulation compared with those of 42.9% and 141 days in cows detected with more than 4 dominant follicles. It is concluded that the number of dominant follicles detected prior to first ovulation varied from I to 5, which positively correlated with ovulation intervals, while only those cows which first ovulated less than 3 dominant follicles resulted in reasonable reproductive performance.     

Reproductive Patterns in the Non‐Breeding Season in Asinina de Miranda Jennies

This study aims to characterize the reproductive patterns in Asinina de Miranda jennies during the non‐breeding season. Reproductive activity was surveyed in 12 females, aged between 3 and 18 years old, using ultrasound and teasing with a jack. The animals were monitored from September to April, six in each consecutive year. Of these 12 females, nine showed disruption to the normal pattern of ovarian activity during the non‐breeding season. Loss of normal cyclicity included anoestrus (41.7%), silent ovulatory oestrus (25%), and persistence of corpus luteum (8.3%). Only three females maintained a regular cyclic pattern with oestrous behaviour during the non‐breeding season. Anoestrus began in early November and lasted for an average of 147 ± 28 days (113–191 days), ending near to the spring equinox. Onset of silent oestrous cycles began more erratically, between October and February. In both groups the first behavioural ovulation of the year occurred around the time of the spring equinox. Disrupted reproductive activity was preceded by a shorter oestrous cycle only in females entering anoestrus. The mean follicle size in the first ovulation of the year was larger than in the reproductive season (44.7 ± 2.45 mm vs 39.2 ± 3.60 mm) in anoestrous jennies with protracted oestrus. Though age and body condition score (BCS) were associated, changes in BCS below a threshold of four points (for anoestrus) and five points (for silent oestrus) contributed greatly to disruption of reproductive cycles. BCS in females with regular oestrous cycles during the winter season remained unchanged or exceeded five points prior to the winter solstice.     

Postpartum follicular development in Brahman cows under two stocking rates

The aim was to study postpartum follicular population dynamics in Brahman cows in the Mexican humid tropics and under different stocking rates. Lactating adult cows (n = 104) grazed native grammas (Paspalum spp., Axonopus spp.) under two stocking rates: 2.8 (high) or 1.7 (low) cows/ha. Follicle numbers and their size (millimetres) were measured by ovarian ultrasound scanning. Animals detected in oestrus and inseminated were taken out of the study. Both stocking rates produced similar (P < 0.05) body condition scores (BCS) and follicular dynamics. The percent of follicles, averaged over stocking rates, of <4 mm diminished and those of 5–8 mm increased with time, whilst those of >9 mm remained constant; respective values for the previously mentioned sizes were 60 days postpartum (n = 101), 49%, 56% and 5%; 120 days (n = 70), 28%, 66% and 6%; and 180 days (n = 15), 9%, 79% and 12%. There was a positive correlation between BCS and follicle number for size <4 mm (r = 0.73, P < 0.01), whilst for size 5–8 mm, this was negative (r = −0.51, P < 0.05), for follicles >9 mm, no difference was found (r = −0.08, P > 0.05). It was concluded that stocking rate did not affect follicular population dynamics of Brahman cows, but BCS dictated the number and categories of follicles present regardless of the stocking rate utilised.     

Expression Pattern of Vascular Endothelial Growth Factor in Canine Folliculogenesis and its Effect on the Growth and Development of Follicles after Ovarian Organ Culture

In this study, the expressions of VEGF in dog follicles were detected by immunohistochemistry and the effects of VEGF treatment on the primordial to primary follicle transition and on subsequent follicle progression were examined using a dog ovary organ culture system. The frozen‐thawed canine ovarian follicles within slices of ovarian cortical tissue were cultured for 7 and 14 days in presence or absence of VEGF. After culture, the ovaries were fixed, sectioned, stained and counted for morphologic analysis. The results showed that VEGF was expressed in the theca cells of antral follicles and in the granulosa cells nearest the oocyte in preantral follicle but not in granulosa cells of primordial and primary follicles; however, the VEGF protein was expressed in CL. After in vitro culture, VEGF caused a decrease in the number of primordial follicles and concomitant increase in the number of primary follicles that showed growth initiation and reached the secondary and preantral stages of development after 7 and 14 days. Follicular viability was also improved in the presence of VEGF after 7 and 14 days in culture. In conclusion, treatment with VEGF was found to promote the activation of primordial follicle development that could provide an alternative approach to stimulate early follicle development in dogs.     

Effects of first postpartum progesterone rise,metabolites, milk yield,and body condition score on the subsequent ovarian activity and fertility in lactating Holstein dairy cows

Seventy multiparous healthy lactating Holstein cows (fat-corrected milk yield = 7,561.8 kg) were monitored from 14 days before to 70 days after calving. Transrectal ultrasound scanning was performed twice weekly from 7 to 65 days postpartum. Blood samples were also collected twice weekly to measure serum P₄ and biweekly to detect serum β-hydroxybutyrate (βHB) and nonesterified fatty acid (NEFA) concentrations. Body condition score (BCS) was taken biweekly after calving. Based on the serum P₄ profile of 59 cows (11 cows were excluded due to the occurrence of postpartum diseases) studied, 27 (45.8%) had normal ovarian activity, while 21 (35.6%), six (10.1%), three (5.1%), and two (3.4%) had delayed ovulation (DOV), prolonged luteal phase (PLP), short luteal phase (SLP), and cessation of ovarian activity, respectively. Cows with PLP had an earlier ovulation compared to that of cows with normal ovarian activity (23.16 ± 4.02 vs 31.9 ± 8.35 days; P < 0.05). PLP cows also had a greater mean ± standard deviation peak milk yield (44.2 ± 5.8 vs 37.2 ± 5.7 kg/day, 75 days postpartum; P < 0.05) than cows with normal postpartum ovarian activity. The PLP group also had greater milk production in the previous lactation period. Logistic regression analysis indicated that cows with P₄ concentration ≥1 ng/ml on day 24 after calving were more at risk for PLP by 1.1 for each 1 kg increase in mean peak milk yield during 75 days after calving. BCS was lower in cows with DOV compared to that of cows with normal ovarian activity at any time after calving (P < 0.05). Serum βHB concentrations in DOV cows were significantly higher than that of normal cows on day 42 after calving (0.69 ± 0.29 vs 0.54 ± 0.12 mmol/L, P < 0.05). No significant difference was found in the mean concentrations of NEFA between cows in different groups at any time after calving (P > 0.05). The concentrations of P₄ on days 28 and 31 were negatively correlated with βHB concentration on day 42 after calving in cows with normal ovarian activity (R = −0.44, P = 0.02). In conclusion, these findings suggest that early ovulation and hence early postpartum P₄ rise in addition to the high milk production could partly be responsible for the occurrence of PLP in dairy cows.     

Ovarian follicular growth, function and turnover in cattle: a review

Studies in cattle assessing changes in number and size of antral follicles, concentrations of estradiol, androgens and progesterone in serum and follicular fluid, and numbers of gonadotropin receptors per follicle during repetitive estrous cycles and postpartum anestrus are reviewed. The rate of growth of small follicles (1 to 3 mm) into larger follicles increases as the estrous cycle progresses from d 1 to 18 (d 0 = estrus). Size of the largest antral follicle present on the ovary also increases with advancement of the estrous cycle. Most large follicles (greater than 10 mm) persist on the ovarian surface for 5 d or more between d 3 and 13 of the bovine estrous cycle. After d 13, most of these large follicles are replaced more frequently by new growing follicles (turnover) with an increased probability for recruitment of the ovulatory follicle after d 18. More research is needed to determine the time required for growth of bovine follicles from small to large antral size and evoke recruitment of the ovulatory follicle. Factors that regulate selection of the ovulatory follicle are unknown but may involve increased frequency of LH pulses in blood, altered blood flow and(or) changes in intrafollicular steroids and proteins. Quantitative evaluation of ovarian follicles indicated occurrence of consistent short-term changes in fluid estradiol and numbers of luteinizing hormone receptors in cells of large follicles only during the pre-ovulatory period. Presumably, low concentrations of follicular estradiol found during most of the estrous cycle are not due to a lack of aromatizable precursor or follicle-stimulating hormone receptors. Follicular fluid concentrations of progesterone increase only near the time of ovulation. Little is known about changes in follicular growth, turnover and function during postpartum anestrus in cattle. However, preliminary data suggest that the steroidogenic capacity of large follicles changes markedly during the postpartum period.     

Ovarian follicular recruitment,granulosa cell steroidogenic potential and growth hormone/insulin-like growth factor-I relationships in suckled beef cows consuming high lipid diets: Effects of graded differences in body condition maintained during the puerperium

The effects of graded differences in body condition score (BCS) on ovarian follicular development, granulosa cell steroidogenic potential, and growth hormone (GH)-insulin-like growth factor-I (IGF-I) relationships were examined during the puerperium in beef cows fed high fat diets. Cows (n = 36) were allocated randomly about 90 d before expected calving to dietary treatments that resulted in BCS of 3 (very thin), 4 (thin), 6 (optimal) and 8 (obese) at parturition. Body weight and BCS were maintained after parturition by adjusting individual intakes of forage throughout the 17-d experiment, and all cows were fed a high lipid supplement (soybean oil; 0.44 kg/500 kg body weight) during this period. Follicular development in BCS 3 and 4 lagged behind that of BCS 6 and 8 throughout most of the study; however, cows in BCS 4 developed numbers of medium and large follicles by day 15 postpartum that were comparable to those of cows in greater BCS. Mean concentrations of total cholesterol (TCH) and total triglycerides (TG) in serum were greater (P < .05) for cows in BCS 8 vs. 3, 4 and 6 through days 12 (TCH) and 16 (TG) postcalving, respectively. High-density lipoprotein cholesterol (HDL-CH), progesterone and estradiol-17β concentrations in follicular fluid were not affected by BCS at ovariectomy (day 17 postpartum). Addition of insulin and low-density lipoprotein (LDL) to culture media maximized (P < .05) production of progesterone by granulosa cells, but in vivo BCS treatments had no effect. Serum GH concentrations were elevated (P < .05) in cows in BCS 3 and 4 compared to those in BCS 6 and 8 throughout the puerperium. Serum IGF-I tended (P < .10) to increase during the first few days after parturition, and both serum and follicular fluid IGF-I increased (P < .02 and P < .001, respectively) with increasing BCS. Data indicate that cows calving in BCS < 4, and fed such that weight and BCS do not increase, are unlikely to respond to short-term dietary fat supplementation.     

Relationships between insulin-like growth factor-I,milk yield,body condition score,and postpartum luteal activity in high-producing dairy cows

The relations between body condition score (BCS), milk yield, serum insulin-like growth factor-I (IGF-I) profile, and luteal activity were investigated in postpartum dairy cows. Seventy-one healthy high-producing multiparous Holstein cows were subjected to transrectal ultrasound scanning twice weekly from the first to the eighth week postpartum. Blood samples were collected twice weekly to measure serum progesterone (P4) and every 2 weeks to detect serum IGF-I concentrations. BCS was monitored weekly after calving. Cows with serum P4 concentrations ≥1 ng/ml on at least two consecutive samplings were considered to have commenced luteal activity. Commencement of luteal activity (C-LA) was observed earlier than 45 days postpartum in 71.8% of cows while 28.2% showed C-LA later than 45 days. Prolonged luteal phase was the most common abnormal pattern of luteal activity observed. Cows with a C-LA earlier than 45 days postpartum had higher (P ≤ 0.05) mean serum concentrations of IGF-I than those with later C-LA. In addition, cows which showed C-LA earlier than 45 days postpartum had more optimal productive indices including shorter calving to conception interval and calving to first service interval (P ≤ 0.05), and fewer services per conception (P = 0.07). C-LA was significantly later in cows that lost more than 0.5 BCS units within 3 weeks postpartum than in those that lost less than 0.5 units BCS during the same interval (P = 0.02). We conclude that high-producing dairy cows with higher postpartum serum IGF-I concentrations have earlier commencement and normal luteal activity, and better reproductive performance. Severity and duration of BCS loss adversely affect commencement of luteal activity.     

Follicular development,oocyte viability and recovery in relation to follicular steroids,prolactin and glycosaminoglycans throughout the estrous period in superovulated heifers with a normal LH surge,no detectable LH surge,and progestin inhibition of LH surge

Estrous cycles of heifers (n = 137) were synchronized with prostaglandin (PGF_2α) and follicular development stimulated with follicle stimulating hormone. Twenty-eight animals were administered Norgestomet implants 12 hr prior to the initial PGF2α injection to suppress the LH surge that initiates ovulation. Animals were ovariectomized every 12 hr after the initial PGF2α (7–9/time, 12–108 hr and at 192 and 240 hr post PGF2α) and divided into three treatment groups to consist of: 1) animals exhibiting a normal luteinizing hormone (LH) surge (n = 86), 2) animals in which no LH surge was detected (n = 23), and 3) suppression of the LH surge via Norgestomet implants (72–108 hr, n = 28). Follicular diameter was measured and follicular fluid was collected for analysis of prolactin, estradiol, progesterone and glycosaminoglycan concentrations. Progesterone concentrations were increased in animals exhibiting an LH surge as compared to animals in which no LH surge was detected; primarily in large follicles (> 8 mm diameter) after the LH surge. Animals not exhibiting an LH surge also had increased follicular progesterone concentrations compared to Norgestomet-implanted animals (242.3 ± 36.3 vs 86.7 ± 6.4 ng/ml, respectively, P < .01), indicating some LH stimulation. Follicular estradiol in animals exhibiting an LH surge increased up to the time of LH surge detection and then declined whereas animals with no LH surge detected had follicular estradiol concentrations that declined after the PGF_2α injection. No differences were noted between those that did not exhibit an LH surge or in which the LH surge was suppressed with Norgestomet in relation to follicular estradiol concentrations. Follicular estradiol concentrations increased with follicular size in all treatment groups (P < .01). Follicular concentrations of prolactin were increased in small follicles (P < .05; ≤ 4 mm diameter) and follicular prolactin increased from 12 to 36 hr post PGF2α injection, then declined after the LH surge. Follicular glycosaminoglycan concentrations decreased with increases in follicular size (P < .01) and were higher in animals that did not exhibit an LH surge (P < .01). No differences in follicular glycosaminoglycans were noted between Norgestomet-implanted animals and those not exhibiting an LH surge. In the animals representing days 4 and 6 of the subsequent estrous cycle (192 and 240 hr post PGF2α), numbers of small-sized follicles were increased. Follicular progesterone and estradiol concentrations were related to atretic large follicles unovulated from the prior estrus and a wave of growth in small and medium follicles. Follicular prolactin and glycosaminoglycans increased with time of the new estrous cycle and were increased in smaller follicles (P < .01). Suppression of LH with progestin implants (Norgestomet) may relate to early effects of progesterone, which may not be totally eliminated at target tissues and subsequently alters the LH surge, steroidogenesis of the follicle, and ovulation. Oocytes were predominantly found in the follicular fluid from animals in which an LH surge was detected and in the buffer wash of follicles in which no LH surge was detected. Oocyte viability was higher in animals exhibiting an LH surge (75% viable) whereas the oocytes of Norgestomet-implanted animals were 75% degenerate.     

Effect of oxygen tension on in vitro viability and development of dog follicles enclosed within the ovarian cortex

Oxygen concentration has been shown to influence in vitro viability and growth of ovarian follicles. The present study examined the effect of oxygen tension on in vitro development of dog follicles enclosed within the ovarian cortex. Ovaries were obtained from domestic dogs (age, 8 months to 2 years), and cortical fragments were recovered. The cortices were then incubated on 1.5% (w/v) agarose gel blocks within a 4‐well culture plate containing Eagle Minimum Essential Medium (MEM). Ovarian follicles within the tissues were processed for histology and assessed for follicle density, viability and diameter immediately after collection (Control) or after 2 or 5 days of in vitro incubation. Apoptotic cells were assessed using TUNEL assay. Comparisons of follicular viability and diameter were performed using analysis of variance followed by Tukey's test (p < 0.05). Comparisons of follicle density and apoptosis among treatments were conducted using Non‐parametric Kruskal–Wallis test followed by Friedman's test (p < 0.05). No difference (p > 0.05) in follicle density was observed among groups at Day 2 of in vitro culture. However, the density of follicles within cortices cultured in 20% oxygen for 5 days significantly reduced compared to the Control and those incubated in 5% concentration. The viability of cultured follicles in all treatments decreased (p < 0.05) compared to the Control after 2 days incubation, and this value further reduced (p < 0.05) in 20% oxygen group at Day 5. There were no differences in the percentages of apoptotic follicles between the two treatment groups (p > 0.05). Nevertheless, after 5 days of culture, the percentage of TUNEL‐positive follicles increased significantly (p < 0.05) in cortices incubated in 20% oxygen environment. In conclusion, our findings demonstrated that 5% oxygen level was superior to 20% concentration in sustaining in vitro viability of dog follicles enclosed within the ovarian cortex.     

Anti-Mullerian hormone and its relationship to ovulation response and fertility in timed AI Bos indicus heifers

This study evaluated the association between plasma anti-Mullerian hormone (AMH) concentration and fertility in Nelore (Bos indicus) heifers submitted to timed artificial insemination (TAI). At the onset of the synchronization protocol, heifers (n = 289) received a subcutaneous P4 ear implant (3 mg) and 2 mg of oestradiol benzoate. Eight days later, the P4 implant was removed and 0.5 mg of oestradiol cypionate, prostaglandin (0.265 mg, i.m.) and equine chorionic gonadotropin (300 UI, i.m.) was administered, and TAI was performed 48 hr after ear implant removal. Ovarian ultrasound evaluations were performed to measure number of ovarian follicles, dominant follicle size and ovulation response. Pregnancy diagnosis was performed by ultrasound 30 days after AI. Heifers with greater circulating AMH had more antral follicles, a smaller dominant follicle near timed ovulation and lower ovulation response to the timed AI protocol compared to heifers with lower circulating AMH. Although AMH and pregnancy outcome had a quadratic-shaped pattern, AMH was not significantly associated with fertility. In conclusion, heifers with lower AMH had larger follicles towards the end of the synchronization protocol and greater ovulation responses, whereas greater circulating AMH was unrelated to conception success.     

Effect of nutrient levels during the far‐off period on postpartum productivity in dairy cows

The present study aimed to determine the effects of differing nutrient levels during the far‐off period on postpartum metabolism and milk production in lactating cows. Twenty‐six multiparous cows were assigned to three dietary treatments in the far‐off period: a low‐energy diet (L, n = 9, 80% intake of the total digestible nutrients requirement), a moderate‐energy diet (M, n = 8, 105%) and a high‐energy diet (H, n = 9, 130%). During the close‐up period, all cows were provided with 105% intake. After parturition, all cows were fed a lactation diet. The BCS recovery was slow, and low milk yield was found in the H group. In the L group, BCS recovery was favorable after parturition, and lactation persistence was increased. The L group had low rumen endotoxin activity and a high initial ovulation rate after parturition. These findings indicate that a high‐energy diet during the far‐off period has a deleterious effect on milk production. In contrast, the restricted diet in the far‐off period increased adaptability with respect to peri‐parturition metabolic changes, improved the post‐parturition nutritional state, and increased milk production. Furthermore, it suggests that the nutrient levels in the far‐off period affect rumen endotoxin activity and reproductive function after parturition.     

The effect of body condition on ovarian activity of free ranging tropical jennies (Equus asinus)

Serial ultrasonography was performed on seven jennies to study the effect of seasonally fluctuating body condition on ovarian activity during the dry, short rainy and long rainy seasons. Kendall Tau correlations and differences of least square mean values were compared. A distinct seasonal pattern was observed both in body condition and ovarian activity. The mean (+/-SD) Body Condition Score (BCS) was lowest during the dry season 2.6 +/- 0.45 when compared with 3.5 +/- 0.52 and 3.9 +/- 0.58 during the short and long rainy seasons, respectively. The mean (+/-SD) number of total follicles was also proportionally low 7.3 +/- 1.6 during the dry season compared with 9.6 +/- 2.4 and 11.3 +/- 3.3 found during the short and the long rainy seasons, respectively. The BCS was positively correlated to the total number of follicles and the diameter of the largest follicle during all seasons. Periods of increasing BCS and transition between seasons with both increasing and decreasing body conditions were strongly correlated to the number and size of ovarian follicles. Seasonal fluctuation in the body condition was found to be the result of variation in feed resource and ovarian activity was also closely following this seasonal pattern. The improvement in body condition has generally, a positive impact on emergence of small growing follicles there by having influence on the total count of ovarian follicles.     

Postpartum follicular and luteal activity in Holstein-Friesian cows genetically selected for high or low mature bodyweight: Relationships with follicle stimulating hormone,insulin, insulin-like growth factor-1 and growth hormone

AIM: To investigate ovarian follicular and luteal activity during the postpartum period of cows genetically selected for high or low mature bodyweight, in relation to metabolic and reproductive endocrine parameters, to determine whether there are differences between strains that could affect fertility outcomes.

METHODS: The presence of follicles ≥5 mm diameter and luteal structures was mapped in the ovaries of 12 high (heavystrain) and 12 low (light-strain) mature bodyweight cows by daily trans-rectal ultrasonography from Day 7 postpartum until the end of their first normal oestrous cycle. Blood samples were collected daily, for measurement of concentrations of follicle stimulating hormone (FSH), progesterone, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and insulin. Intervals to first ovulation were calculated from ultrasonography data.

RESULTS: Heavy-strain cows had shorter intervals than light-strain cows from calving to the emergence of the first (9.0 (SE 0.9) vs 12.4 (SE 1.3) days) and second (16.4 (SE 1.8) vs 20.6 (SE 1.6) days) dominant follicles (p<0.05). Concentrations of FSH in heavy-strain cows prior to the emergence of the second, third and fourth dominant follicles were higher than in light-strain cows (p<0.05). Heavy-strain cows were more likely to have a large (>15 mm diameter) follicle earlier than light-strain cows (p<0.01). Concentrations of insulin and IGF-1, but not those of GH, were higher in heavy- than light-strain cows during the postpartum period (p=0.01 and p=0.02, respectively), and concentrations of both on Day 6 were inversely related to the time of emergence of the first dominant follicle (p>0.01).

Concentrations of progesterone were similar in both strains of cow until Day 10 of the first oestrous cycle, but thereafter were higher in light- than heavy-strain cows until Day 16. Progesterone concentrations in heavy-strain cows declined earlier and more rapidly than in their lighter counterparts.

CONCLUSION: These results indicate that there is a rapid postpartum resumption of follicular activity in both heavy-and light-strain cows, but that there is an earlier emergence of dominant follicles and ovulation in the former. Differences in luteal function, in terms of lower dioestrus progesterone concentrations and an earlier onset of luteolysis, in heavy- than light-strain cows might be sufficient to impair the fertility of the former.