Systemic profiling of ectopic fat deposits in the reproductive tract of dairy cows.

During the transition period, high-yielding dairy cows suffer from negative energy balance, intense lipomobilization and impaired lipid metabolism; this metabolic condition can lead to overburdened triglycerides accumulation in the liver, known as liver lipidosis, which has been associated to impaired fertility in dairy cows. The mechanisms of this impairment can be in principle correlated with the presence and the extent of ectopic fat depots. However, current methods for evaluating fat accumulation in liver and in the reproductive tract suffer from low resolution, sensitivity, and specificity. Confocal microscopes are equipped with Gallium arsenide phosphide detectors, thus enabling the acquisition of intense signals from tissue biopsies. This method could differentiate whether fat deposition occurred without requiring sample sectioning. Here, we examined with this technique liver, uterine and ovarian samples of heifers and regularly slaughtered repeat breeder and overconditioned dairy cows, to quantify lipid droplets and depots at a submicrometer scale with high specificity. With the aid of this technique, we found lipid depots in uterine and ovarian specimens. Moreover, we found that the size and number of depots increased with the degree of liver lipidosis. Further studies are needed to elucidate the relationship between the severity and extent of these deposits and the fertility of lactating dairy cows. Since tissues other than liver display different characteristic lipid droplet distributions, this technique can be potentially employed to shed new light on the pathogenesis of lipidosis and to assess new risk factors for infertility.

Influence of lipoproteins at dry-off on metabolism of dairy cows during transition period and on postpartum reproductive outcomes.

High-yielding dairy cows are metabolically challenged during transition, when intense mobilization and hepatic oxidation of lipids is achieved, thus leading to fatty infiltration, ketosis and generalized inflammation. The condition is associated to periparturient diseases and poor fertility. The aim of this study was to assess whether serum lipoprotein concentrations in the dry period could influence the occurrence of postpartum diseases and reproductive performance in dairy cows. The study was carried out on 30 multiparous Holstein Friesian cows. Blood samples were collected at dry-off (-60 days), 30 days after dry-off and within 12 h after parturition for biochemical and serum lipoprotein assays. From 10 to 60 days after parturition milk was collected twice weekly after feeding, for milk whey progesterone assay. The Optimal Cutpoint package identified a threshold of 89% for serum High Density Lipoprotein (HDL) concentration at the beginning of the dry period with 95% of confidence interval. Cows with serum HDL greater than 89% (High group, n = 10) showed better reproductive performance when compared to those with low values (Low group, n = 18). The odds ratio for reproductive disorders in High group was 0.6875, however, differences were not significant probably due to both the reduced number of animals per group and overall low incidence of postpartum reproductive disease. First postpartum luteal activity occurred around day 23, while the second one between days 40 and 48. The average calving to first AI interval was 64.00 ± 3.95 days and 94.50 ± 12.32 days in High and Low group, respectively (P < 0.05). The calving-conception interval was 129.86 ± 24.42 days and 199.18 ± 24.73 days in High and Low groups, respectively (P < 0.05). Low group displayed an increase in liver markers, that is total bilirubin, with 0.46 ± 0.09 mg/dL and 0.23 ± 0.08 mg/dL, in Low and High group respectively (P < 0.05), and NEFA/cholesterol ratio, with 0.30 ± 0.06 and 0.14 ± 0.03, in Low and High groups, respectively (P < 0.05), at parturition. Concentrations of HDL >89% at dry-off could be suggestive of improved liver adaptation to the transition, and probably of enhanced fertility in High group.

Reproductive performance of dairy cows with luteal or follicular ovarian cysts after treatment with buserelin.

In dairy farm management economic losses resulting from cystic ovarian degeneration are well known. In spite of this, neither the definition nor the aetiopathology of ovarian cysts are clear and agreed upon. Also the usual classification in luteal and follicular cysts, requiring ultrasound examination together with assessment of P4 to be accurate, is not very helpful in field conditions. Consequently a single treatment is often provided for both types of cysts, and since the 1970s treatments with GnRH and its analogues have been considered very useful. Nevertheless differences in recovery rates after GnRH treatment in animals with either luteal or follicular cysts are reported. Thus, the aim of this study was to evaluate recovery rate, recovery time and conception rate after treatment with buserelin (GnRH-analogue) in cows with ovarian luteal or follicular cysts. In a 5 years period, 150 cows with cysts out of a total of 990 animals, were detected and treated intravenously between 45 and 60 days PP with 20µg buserelin. No statistically significant differences were found in recovery rates and in conception rates between the two types of cysts. Comparison of recovery times showed significantly shorter recovery for cows with luteal cysts. The results emphasise the usefulness of GnRH to treat ovarian cysts regardless of their type, in relation to both recovery and conception rates. Intervals from treatment to resumption of ovarian activity were affected by the characteristics of ovarian cysts, with a faster recovery for the luteal type.

Gonadotropin secretion and pituitary responsiveness to GnRH in mares with granulosa-theca cell tumor.

Granulosa-theca cell tumors (GTCTs) are able to secrete variable amounts of sex steroids and immunoreactive inhibin (ir-INH). Although the pituitary appears to be affected by the presence of a GTCT, pituitary responsiveness to exogenous GnRH has not been examined. The aims of the present study were to: (i) assess the plasma hormone concentrations of ir-INH, gonadotropins and sex steroids in eight mares with GTCT and (ii) assess the responsiveness of pituitary gonadotroph cells to exogenous GnRH stimulus both before and after tumor removal. In seven mares, the contralateral ovary was firm, small and inactive. Histopathological observations of the tumors confirmed the presumptive diagnosis of a GTCT. Four mares, judged to be in vernal transition period (n=2) and in the breeding season (n=2), were used as controls. A single intravenous injection of 40 microg of GnRH agonist was given to each mare and blood samples were collected every 15 min from 2 h before to 4 h after injection. In four GTCT mares, this procedure was repeated 20 (n=2) and 90 (n=2) days after tumors removal. All plasma samples were analyzed for concentrations of ir-INH, LH, FSH, estradiol-17beta (E2), testosterone (T) by RIA and progesterone (P) by EIA. Results showed that E2 levels were significantly higher (P<0.001) in control animals compared to E2 levels in GTCT mares before and after surgery. P and T concentrations were not statistically different between the groups. Baseline levels of ir-INH were greater (P<0.05) in GTCT mares before surgery than in control mares, and decreased to undetectable levels after neoplasia ablation. Baseline FSH did not differ between control and GTCT animals either before or after the ovaries were removed. LH baseline values appeared to be higher for affected mares, but the difference was not statistically significant. Maximum release (MR) and area under the gonadotrophin release curve (AUC) after the GnRH challenge for both the gonadotrophins were similar between the groups.