Comparison of serum concentrations of pregnancy-associated glycoproteins and estrone sulphate during pregnancy in eutocia and dystocia beef cattle.

Preventing dystocia can stabilise beef cattle management. This study aimed to investigate the relationship between serum pregnancy-associated glycoproteins (PAGs) S-N values and estrone sulphate (E1S) concentrations during pregnancy and the calf birth weight in beef cattle and to evaluate their usefulness as new predictive parameters for dystocia due to foetal overgrowth. Thirty-eight pregnant Japanese Black cattle were used. Blood samples were collected at 40, 70, 100, 150, 200, 250, 280, and 285 days after artificial insemination (AI), and birth weight of the offspring was measured. Serum PAGs S-N values and E1S concentrations were measured, and the area under the curve (AUC) and the ratio of change based on 70 days after AI were calculated, followed by calculation of the correlation coefficient with the birth weight of the offspring and comparison between the eutocia (n = 32) and dystocia (n = 6) groups. The birth weight of the offspring was moderately positively correlated with the AUC of serum PAGs S-N values and E1S concentrations in the second (r = 0.425, P < 0.01) and third (r = 0.595, P < 0.01) trimesters, respectively. The ratio of change in serum E1S concentrations between 70 and 280 days after AI was greater (P < 0.05) in the dystocia group (1276.6 ±â€¯229.1 %) than in the eutocia group (852.6 ±â€¯69.6 %). These results suggest that blood PAGs S-N values at mid-pregnancy (100-199 days after AI) and the ratio of changes in blood E1S concentrations between 70 and 280 days after AI may be new parameters for predicting dystocia.

Establishment of a method to predict dystocia due to physical imbalance between foetus and maternal pelvis in Japanese black cattle.

This study aimed to clarify the relationship between the maternal pelvis and calf size in normal and dystocia groups (Experiment 1) and to establish a method to predict dystocia due to physical imbalance between foetus and maternal pelvis before calving based on the parameters found in Experiment 1 (Experiment 2). In Experiment 1, 26 pairs of dams and their offspring were tested immediately after calving, and various parameters of offspring size and maternal pelvis were measured in normal and dystocia cases. The longitudinal diameter of the maternal pelvis was smaller (p < .01), and ratios of the offspring's birth weight and the width of the hoof crown (hoof width; measured using callipers) to the longitudinal diameter of the maternal pelvis were larger (p < .01) in the dystocia group than those in the normal group. In Experiment 2, 37 cows were tested, and in addition to the measurement items of Experiment 1, the inner pelvic diameter and the foetal hoof width were measured via ultrasonography at 285 days of gestation. The ratio of the internal diameter of the maternal pelvis (longitudinal diameter, transverse diameter and area) to the foetal hoof width was compared and was greater (p < .01) in the dystocia group than in the normal group. In conclusion, the risk of dystocia can be estimated by the ratio of the foetal hoof width to the longitudinal diameter of the maternal pelvis before calving.

Association of interservice interval with conception rate in Japanese Black cattle.

This study aimed to clarify the association of interservice interval (ISI) with conception rate (CR) and to evaluate factors affecting ISI in Japanese Black cattle. Data used in this study covered 32,639 artificial insemination (AI) records on 972 farms. The mean ± SEM of ISI and CR was 57.0 ± 0.3 days and 44.6 ± 0.3%, respectively. The relative frequency of ISI differed significantly between parity groups (p < .05). For parity 0, the proportion of ISI at an 18- to 24-day interval was 43.6%, whereas the proportions in the other parity groups ranged from 27.2% to 29.1%. The CR was associated significantly with ISI, parity and AI number (p < .05), but not with season. Cows reinseminated at an 18- to 24-day interval had the highest CR (46.8%; p < .05). Cows reinseminated at a 39- to 45-day interval had a similar CR to those reinseminated at 25- 38-day and 46- to 59-day intervals. Cows reinseminated at 11-17, 60-66 and 67 days or longer intervals had the lowest CR (p < .05). Thus, cows returning to oestrus in a normal cycle tended to have a higher probability of conceiving compared with those that returned to oestrus after a normal cycle for any degree of parity and at any number of AI.

Characterization of anti-Müllerian hormone in a case of bovine male pseudohermaphroditism.

The current report aimed to characterize plasma anti-Müllerian hormone (AMH) in bovine male pseudohermaphroditism. The blood AMH concentration in a Japanese Black male pseudohermaphrodite calf was compared with pre- and post-pubertal male and female calves and castrated calves. The concentration in the case was higher than in post-pubertal males, castrated males, and pre- and post-pubertal female calves (p < .05), but similar to that in pre-pubertal male calves. After extraction of the testes, the concentration in the case dropped to a certain extent. The extracted testes expressed AMH, as detected by immunohistochemistry. This study is the first to show the characterization of AMH in a male pseudohermaphrodite calf. AMH levels in peripheral blood might be useful to diagnose male pseudohermaphroditism in cattle.

Histological and immunohistochemical evaluation of granulosa cells during different stages of folliculogenesis in bovine ovaries.

Bovine granulosa cells (GC) vary in their morphological aspect during different stages of folliculogenesis. In this study, 10 morphologically normal bovine ovaries were collected to study the structural aspects of different stages of GC using intermediate filament protein antibodies including cytokeratin AE1/AE3 (AE1/AE3), vimentin, nectin-4 and desmin. Hormonal immunolocalization was assessed using the immunomarkers anti-Müllerian hormone (AMH) and inhibin alpha. In addition, tumour markers and proliferation markers using c-erbB-2 oncoprotein and proliferating cell nuclear antigen, respectively, were investigated. The immunolabelling of AE1/AE3 in GC was strongest in the early follicle stage and gradually decreased when reaching the Graafian follicle stage. Its immunolabelling increased again as the stage progressed from stage I to stage III. The immunolabelling of inhibin alpha was inversely proportional to that of AE1/AE3 in the developing ovarian follicles as their immunolabelling is opposite to each other during folliculogenesis. AMH was immunopositive in almost all GC stages in different intensities and percentages, except for some negative staining in the atretic IV follicles. The atretic IV follicle is a unique type of atretic follicle that shows Call-Exner body formation, which was mainly found in older cows in this study. The distinct patterns of immunoreactivity for various types of immunomarkers in the different GC stages will play an important role in diagnostic assistance of various follicle conditions, including cystic ovaries and GC tumours.

Effects of service number on conception rate in Japanese Black cattle.

The conception rate (CR) of Japanese Black cattle has been decreasing since 1990. The objectives of this study were to compare CR by artificial insemination (AI) number, and to assess the interaction between AI number and possible factors related to CR in Japanese Black cattle. Records of 11,182 AI records for 6,741 heifers and 61,302 AI records for 13,139 cows on 977 farms were analysed. The average CR of heifers was 47.0%, and CR at the first, second and third AI was higher than at the fourth or subsequent AI (p < 0.05). The average CR of cows was 47.8%, and their CR at first and second AI was higher than the fourth or subsequent AI (p < 0.05). Data analysis with CR as the dependent variable revealed significant interactions of AI number with interval from calving to first AI and AI season. Cows first serviced at ≤48 days post-partum had lower CR than those at ≥90 days at first AI (p < 0.05), but the interval from calving to first AI did not affect CR at second AI. Cows serviced in the autumn had a higher CR at first AI than those serviced in the spring and winter (p < 0.05); however, there was no seasonal difference in CR at the second AI. In summary, CR began to decrease from the fourth AI in heifers and the third AI in cows. The AI season and the interval from calving to first AI significantly affected CR only at the first AI.

Rapid Communication: MiR-92a as a housekeeping gene for analysis of bovine mastitis-related microRNA in milk.

Our aim was to identify a suitable microRNA housekeeping gene for real-time PCR analysis of bovine mastitis-related microRNA in milk. We identified , , and as housekeeping gene candidates on the basis of previous Solexa sequencing results. Threshold cycle (CT) values for , , and did not differ between milk from control cows and milk from mastitis-affected cows. NormFinder software identified as the most stable single housekeeping gene. We evaluated the suitability of the housekeeping gene candidates by using them to assess expression levels of the inflammation-related gene . Regardless of the housekeeping gene candidates used for normalization, relative expression levels of were significantly higher in mastitis-affected samples than in control samples. However, of all the housekeeping genes and gene combinations investigated, normalization with alone generated the difference in relative expression between mastitis-affected and control samples with the highest significance. These results suggest that is suitable for use as a housekeeping gene for analysis of bovine mastitis-related microRNA in milk.

Relationship between plasma anti-Müllerian hormone concentrations during the rearing period and subsequent embryo productivity in Japanese black cattle.

To use plasma anti-Müllerian hormone (AMH) concentrations as early-stage markers for donor cow selection, we investigated the relationship between plasma AMH concentrations in Japanese black heifers and subsequent embryo productivity following superovulation treatment. Plasma AMH and nonesterified fatty acid (NEFA) concentrations in 6 heifers were evaluated once per month from 3 mo before successful AI for primiparity to 3 mo postpartum. Following calving, embryo collection by superovulation treatment was performed at 3- to 4-mo intervals. There were no significant differences in plasma AMH concentrations between the time points throughout the study period. There were, however, significant inter-animal differences in plasma AMH concentrations (P < 0.05). These findings suggest that plasma AMH concentrations were stable over time and individually specific. There were significant positive correlations between plasma AMH concentrations before AI and embryo productivity variables, including the number of ova/embryos (number of transferable embryos, degenerated embryos, and unfertilized oocytes) and numbers/proportions of fertilized and transferable embryos. There was no significant correlation between plasma AMH and NEFA concentrations throughout the study period. These findings reveal that plasma AMH concentrations during the rearing period can be used to predict subsequent embryo productivity following superovulation treatment, suggesting that these concentrations are useful early-stage markers for selecting donor cows.

Changes in peripheral anti-Müllerian hormone concentration and their relationship with testicular structure in beef bull calves.

The aim of this study was to clarify the time-course of changes in anti-Müllerian hormone (AMH) and testosterone (T) concentrations in peripheral blood and to determine the relationships between blood AMH concentration and testicular development during the early postnatal and prepubertal periods in beef bull calves. A total of 17 Japanese Black bull calves were enrolled in this study. The wk in which the calf was born (within 6 d after birth) was defined as M 0. Blood samples were taken once in every mo from M 0 to M 6 from each bull calf, and plasma AMH and T concentrations were determined. Of the 17 calves, 10 were castrated at 6 mo of age (prepuberty) and the right testis was histologically examined. Plasma AMH concentration (means ± SE) at M 0, 1, and 2 were 123.5 ± 9.8, 189.6 ± 18.7, and 254.6 ± 14.1 ng/mL, respectively. From M 0 through M 2, plasma AMH concentration was significantly greater each mo than in the previous mo (P < 0.05); however, plasma AMH concentration significantly decreased over the last 3 mo of the study (P < 0.05). The average age at which plasma AMH concentration was the highest was 2.3 ± 0.1 mo of age. Plasma T concentration significantly increased from M 0 (0.18 ± 0.02 ng/mL) until M 6 (6.52 ± 1.41 ng/mL). Plasma AMH and T concentrations at M 4, 5, and 6 were significantly negatively correlated (P < 0.05). Linear regression did not reveal a significant relationship between Sertoli or Leydig cell numbers and plasma AMH or T concentrations, respectively. In conclusion, blood AMH concentration peaks at 2 mo of age and is negatively correlated with blood T concentration from 4 to 6 mo of age. Although prepubertal blood AMH or T concentrations did not reflect Sertoli or Leydig cell numbers at the end of the prepubertal period, blood AMH concentration may be indicative of abnormal Sertoli cells function.

Evidence of Spontaneous Recovery of Granulosa-Theca Cell Tumour in a Heifer: A Retrospective Report.

A 14-month-old Japanese Black heifer was evaluated on Day 0 (D 0) for enlargement of the right ovary (RO). Transrectal ultrasonography (TRUS) revealed that the RO was markedly enlarged and multicystic, while the left ovary (LO) was small and inactive. The presumptive diagnosis was granulosa-theca cell tumour (GTCT), which was confirmed by markedly elevated plasma anti-Müllerian hormone (AMH) of 4.42 ng/ml. Therefore, ovariectomy of the RO was the treatment of choice. The heifer was checked by TRUS and blood sampling on D 42, D 63 and immediately before ovariectomy on D 85. On D 42, TRUS did not show marked changes in either ovary in comparison with D 0. However, on D 63, the RO had transformed into a single cyst, and on D 85, the LO had resumed cyclic activity. The RO was extracted on D 85 by hand-assisted laparoscopic ovariectomy to allow better control. Unexpectedly, histopathology revealed the lesion to be a fluid-filled cystic structure, with no neoplastic proliferation of follicular epithelium that would indicate GTCT. The wall of the cystic structure consisted of collagen fibres and a few degenerated granulosa cells. The retrospective hormonal analysis revealed that the AMH concentrations had markedly dropped on D 63 and 85, which coincided with resumption of cyclicity in the LO. These findings suggest that the GTCT had self-cured and transformed into a cyst-like structure. The heifer then received an oestrous synchronization regime on D 105, was artificially inseminated on D 115 and became pregnant.