In vitro embryo production in buffalo: current situation and future perspectives

In the last few years, there has been an increasing interest in the in vitro embryo production technologies for faster propagation of superior germplasm in buffalo, due to the low efficiency of multiple ovulation and embryo transfer programs. Early attempts to produce buffalo embryos in vitro have been made by using procedures that were proven effective in cattle. However, the acquisition of more specific information on oocyte and embryo culture requirements in vitro in this species has resulted in an improved efficiency over the years. Although the IVEP efficiency has enhanced, as indicated by competitive embryo yields, pregnancy rate and development to term are still poor. Furthermore, the optimization of embryo cryopreservation methods in this species is critical for the diffusion of ET procedures in the field. The present review intends to describe the state of the art of IVEP in buffalo species, emphasizing the advances achieved and the limitations still to overcome

A penalized framework for distributed lag non-linear models.

: Distributed lag non-linear models (DLNMs) are a modelling tool for describing potentially non-linear and delayed dependencies. Here, we illustrate an extension of the DLNM framework through the use of penalized splines within generalized additive models (GAM). This extension offers built-in model selection procedures and the possibility of accommodating assumptions on the shape of the lag structure through specific penalties. In addition, this framework includes, as special cases, simpler models previously proposed for linear relationships (DLMs). Alternative versions of penalized DLNMs are compared with each other and with the standard unpenalized version in a simulation study. Results show that this penalized extension to the DLNM class provides greater flexibility and improved inferential properties. The framework exploits recent theoretical developments of GAMs and is implemented using efficient routines within freely available software. Real-data applications are illustrated through two reproducible examples in time series and survival analysis.<br/

Metabolic profile in growing buffalo heifers fed diet with different energy content

Aim of this study was to verify the relation among the mediators and indicators of nutritional status like insulin, glucagon, urea, cholesterol, triglycerides and total proteins in growing buffalo heifers, fed diets with different energy density. 12 Murrah heifers were randomly allocated into two dietary treatments (High, Group H; Low, Group L) that differed in energetic levels (Group H: 5.8 UFL/d; Group L: 3.6 UFL/d). Every 30 days, for a total of five times, blood samples were collected at 08.00 h, before feeding, from the jugular vein in vacutainer tubes and analysed to determine metabolic profile. Data on haematic constants were analysed by ANOVA for repeated measures with treatment as the main factor. Low energy availability and low NSC reduced the glucose and insulin and increased glucagone and urea blood levels. The increase of NSC in the diet of group H during the experiment may caused a reduction of the fibre digestibility after the period of adaptation of the rumen microflora and, as a paradox effect, suffered for an energetic lack with a subsequent activation of lipolysis and mobilization of their body reserves. Liver and muscular synthesis increase in group with a high energy availability

Relationship of body condition score and blood urea and ammonia to pregnancy in Italian Mediterranean buffaloes

The relationship of body condition score ( BCS) and blood urea and ammonia to pregnancy outcome was examined in Italian Mediterranean Buffalo cows mated by AI. The study was conducted on 150 buffaloes at 145 +/- 83 days in milk that were fed a diet comprising 14.8% crude protein, 0.9 milk forage units . kg(-1) dry matter and a non- structural carbohydrate/ crude protein ratio of 2.14. The stage of the oestrous cycle was synchronised by the Ovsynch- TAI programme and blood urea and ammonia levels were assessed on the day of AI. Energy corrected milk ( ECM) production and BCS were recorded bi- weekly. The pregnancy risk was 46.7% and was slightly lower in buffaloes with BCS 7.5. There were no significant differences in ECM, urea and ammonia between pregnant and non- pregnant buffaloes. However, pregnancy outcome was higher ( P = 0.02) in buffaloes with blood urea < 6.83 mmol . L-1. The likelihood of pregnancy for buffaloes with low urea blood level was 2.6 greater than for high urea level and exposure to a high urea level lowered the probability of pregnancy by about 0.25. The findings indicate that buffaloes are similar to cattle and increased blood levels of urea are associated with reduced fertility when animals are mated by AI

Seasonal effects on miRNA and transcriptomic profile of oocytes and follicular cells in buffalo (Bubalus bubalis)

Season clearly influences oocyte competence in buffalo (Bubalus bubalis); however, changes in the oocyte molecular status in relation to season are poorly understood. This study characterizes the microRNA (miRNA) and transcriptomic profiles of oocytes (OOs) and corresponding follicular cells (FCs) from buffalo ovaries collected in the breeding (BS) and non-breeding (NBS) seasons. In the BS, cleavage and blastocyst rates are significantly higher compared to NBS. Thirteen miRNAs and two mRNAs showed differential expression (DE) in FCs between BS and NBS. DE-miRNAs target gene analysis uncovered pathways associated with transforming growth factor \u3b2 (TGF\u3b2) and circadian clock photoperiod. Oocytes cluster in function of season for their miRNA content, showing 13 DE-miRNAs between BS and NBS. Between the two seasons, 22 differentially expressed genes were also observed. Gene Ontology (GO) analysis of miRNA target genes and differentially expressed genes (DEGs) in OOs highlights pathways related to triglyceride and sterol biosynthesis and storage. Co-expression analysis of miRNAs and mRNAs revealed a positive correlation between miR-296-3p and genes related to metabolism and hormone regulation. In conclusion, season significantly affects female fertility in buffalo and impacts on oocyte transcriptomic of genes related to folliculogenesis and acquisition of oocyte competence

Effects of dietary supplementation of conjugated linoleic acids and their inclusion in semen extenders on bovine sperm quality

Isomers of conjugated linoleic acid (CLA) enhances circulating insulin-like growth factor I (IGF-I) levels. Furthermore, fertility rate of breeding bulls is positively correlated to seminal plasma IGF-I concentration. Our objective was to evaluate the effect of dietary CLA supplementa-tion and inclusion to the semen extender on bovine semen quality and freezability. Fourteen bulls, randomly assigned to control (CTL) and CLA (50 g/day) groups, were supplemented for 10 weeks. Samples were collected at Weeks −2 (before supplementation), 0, 4, 6 (during supplementation), 10, and 11 (after supplementation). Blood and seminal plasma were analyzed for IGF-I; the ejaculates were frozen in the following subgroups: CTL (no addition to semen extender), CLA c9, t11 (50 µM), CLA c9, t11 (100 µM), CLA t10, c12 (50 µM), CLA t10, c12 (100 µM), and CLA mix (50 µM each of CLA c9, t11 and CLA t10, c12). Sperm motility, morphology, viability, mitochondrial membrane potential, and reactive oxidative species were assessed. CLA supplementation decreased ejaculates’ total volume, increased sperm concentration, beat cross frequency, and decreased oxidative stress; it also increased plasma and seminal plasma IGF-I levels compared to the CTL. The inclusion of CLA c9, t11 100 µ M and CLA mixture in the extender increased live spermatozoa percentage post-thawing compared to other groups. Our results show a beneficial effect of CLA supplementation on semen quality; however, further studies evaluating fertilization rates are necessary to corroborate the results

Estimation of buffalo cheese yield by using the chemical-physical parameters of the milk

The aim of this study was to estimate cheese yield by using the chemical- physical parameters of the milk. Analysis were performed on 325 milk samples with 80-219 days in milk interval. Furthermore, buffaloes which showed a ratio between theoretical cheese yield (calculated by Altiero formula) and real cheese yield at 28 hours higher (Group A) or lower (Group B) than 0.983, were compared taking into account 5 hypothetical analytical potentialities of laboratories: 1) Fat percentage; 2) Protein and fat percentages; 3) Protein and fat percentages, pH and SH; 4) Protein and fat percentages, pH, SH, urea, protein percentage corrected per urea, lactose, solids-not-fat (SNF) and SCC; 5) Protein and fat percentages, pH, SH, urea, protein percentage corrected per urea, lactose, SNF, SCC, TAMF, milk DM percentage, ash percentage and casein percentage. Correlation and regression analyses with stepwise method were performed for curd quantity in relation to the physic-chemical ad microbiological milk composition by using SPSS 15.0. As expected, R2 value was such high as the number of variables included in the calculation. A higher R2 value was observed in those samples characterized by a ThCY/28CY ratio < 0.983. ThCY calculated according to Altiero et al (1989), underestimated 28CY of +1.8 g/litre in all samples, whereas a difference between –2.2 (Laboratory 2) and +1.0 (Laboratory 3) g/litre was registered if the actual formula is utilized. According to Altiero formula, 28CY was overestimated of 9.6 g/litre in Group A, whereas it was underestimated of 1.8 g/litre in Group B. According to our study, the estimation of 28CY showed a difference between –9.3 (Laboratory 2) and 9 (Laboratory 1) g/litre in Group A and – 3.5 (Laboratory 1) e 0.0 (Laboratory 5) g/litre