Equine Chorionic Gonadotropin as an Effective FSH Replacement for In Vitro Ovine Follicle and Oocyte Development.

The use of assisted reproductive technologies (ART) still requires strategies through which to maximize individual fertility chances. In vitro folliculogenesis (ivF) may represent a valid option to convey the large source of immature oocytes in ART. Several efforts have been made to set up ivF cultural protocols in medium-sized mammals, starting with the identification of the most suitable gonadotropic stimulus. In this study, Equine Chorionic Gonadotropin (eCG) is proposed as an alternative to Follicle Stimulating Hormone (FSH) based on its long superovulation use, trans-species validation, long half-life, and low costs. The use of 3D ivF on single-ovine preantral (PA) follicles allowed us to compare the hormonal effects and to validate their influence under two different cultural conditions. The use of eCG helped to stimulate the in vitro growth of ovine PA follicles by maximizing its influence under FBS-free medium. Higher performance of follicular growth, antrum formation, steroidogenic activity and gap junction marker expression were recorded. In addition, eCG, promoted a positive effect on the germinal compartment, leading to a higher incidence of meiotic competent oocytes. These findings should help to widen the use of eCG to ivF as a valid and largely available hormonal support enabling a synchronized in vitro follicle and oocyte development.

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.

Validation of the Italian version of the UNESP-Botucatu unidimensional composite pain scale for the assessment of postoperative pain in cattle.

OBJECTIVE: The aim of this study was to validate the Italian version of the UNESP-Botucatu Unidimensional Composite Pain Scale (UCPS-IV) for assessing postoperative pain in cattle. STUDY DESIGN: Video analysis and psychometric testing. ANIMALS: A total of 40 Nellore cattle, age 2-3 years, weighing 365±51 kg. METHODS: An English version of the scale was translated to Italian and back translated to English to ensure semantic equivalence. A total of four Italian observers and the researcher who developed the original scale (considered as the gold standard) analysed videos of 40 cattle previously subjected to orchiectomy (recorded for each cattle at four predetermined timepoints during the perioperative period) unaware of the videos' timepoint. They assigned a pain score [ranging from no pain (0) to severe pain (10)] using the Italian version of the scale and verified the need for analgesic treatment for each animal. The obtained scores were subjected to psychometric validity, responsiveness and reliability tests. RESULTS: The factor analysis of the scale resulted in a one-factor solution. Significant changes in pain scores in response to surgery and analgesics confirmed not only the content and construct validity, but also responsiveness. The agreement between the researcher who developed the original scale and the blinded observers and the correlation between the pain scores determined by the scale and those determined by three classical unidimensional pain scales confirmed criterion and concurrent validity. Internal consistency of the scale was excellent. Inter- and intrarater reliability ranged from moderate to good and from moderate to very good for all scale items, respectively, supporting reproducibility and stability. The cut-off point for rescue analgesia calculated by the receiver operating characteristic curve was > 3. CONCLUSIONS AND CLINICAL RELEVANCE: The results confirm that the UNESP-Botucatu UCPS-IV appears to be a valid and reliable tool for the assessment of postorchiectomy pain in the bovine species. The determination of a cut-off point for rescue analgesia is also helpful for guiding analgesic therapy.

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.