Differential expression of insulin‐like growth factor family members in immature cumulus–oocyte complexes from dairy cows with different genotypes

It has been evident the improvement of in vitro embryo production (IVEP) in dairy cows. Nevertheless, it is known that differences in the number and quality of oocytes between taurine and zebu females impact the efficiency and economic viability of IVEP. As the insulin‐like growth factor (IGF) system is related to follicular and oocyte development, we aimed to quantify mRNA abundance of IGF system members and pregnancy‐associated plasma protein‐A (PAPPA) in the cumulus–oocyte complexes (COCs) of Gir, 1/2 Holstein × 1/2 Gir and Holstein cows. Four pools of 30 immature COCs from Gir, 1/2 Holstein × 1/2 Gir and Holstein cows were obtained by ovum pickup (OPU), and the oocytes and cumulus cells (CC) were mechanically separated and stored at ?80°C. Total RNA was extracted from pools of 30 oocytes and their respective CC. Expression of target genes was assessed by real‐time RT‐PCR. In oocytes, the abundance of IGFR1 mRNA was higher (p < .05) in Gir cows compared with the other breeds. In contrast, in CC, mRNA encoding IGF2 (p < .05), IGFR2 (p < .05) and IGFBP4 (p < .01) was higher in Holstein donors compared with Gir and 1/2 Holstein × 1/2 Gir cows. Additionally, the abundance of PAPPA mRNA was higher in oocytes (p < .001) and CC (p < .01) in Gir and 1/2 Holstein × 1/2 Gir cows compared with the Holstein donors. In conclusion, the higher abundance of PAPPA mRNA in the oocytes and CC from Gir and cross‐breed donors combined with the low expression of IGFBP4 in the CC suggests an enhancement of the bioavailability of IGF‐free when compared with Holstein COCs.     

Infection of cattle in Queensland with bluetongue viruses: 1. prevalence of antibodies

SUMMARY A survey of nearly 20 000 cattle in Queensland was conducted to describe the prevalence and distribution of infection by serotypes of bluetongue virus. The overall prevalence of serum antibodies to one or more bluetongue viruses was 8.7% (95% confidence interval 8.3 to 9.1). Sera from cattle contained neutralising activity against 2 serotypes, 1 and 21. No evidence was found of infection with other serotypes previously isolated in Australia. The overall prevalence of serotype 1 antibodies was 7.7% (95% CI 7.3 to 8.0) and the prevalence of serotype 21 antibodies was 3.3% (95% CI 3.1 to 3.6). The prevalence of serotype 1 antibodies was significantly (P < 0.05) higher than that of serotype 21 in every region of the State, except in the central highlands and south-west Queensland. Overall, 3 significantly (P < 0.05) different zones of prevalence were found: high prevalence (> 20%) in far north Queensland, moderate (5 to 20%) in north-west, northern and southern coastal Queensland, and low (< 5%) in the central highlands, Darling Downs and south-west Queensland.     

Regulation of Corpus Luteum-function in the Bitch

Functional lifespan of the corpus luteum (CL) in non-pregnant dogs exceeds that of pregnant animals and may last for more than 80 days. Prolactin and LH act luteotropic, however, luteolytic mechanisms are poorly understood. Other than in life stock there is no uterine luteolysine and it was postulated that local paracrine/autocrine mechanisms might play a major role. In following this hypothesis the present investigations have clearly demonstrated that up-regulation of prostaglandin synthesis in the CL as indicated by the expression of cyclo-oxygenase II occurs with its formation and not regression, pointing towards a luteotropic rather than luteolytic action. Throughout dioestrus luteal and other cells of the CL express the oestrogen (ERalpha) and progesterone receptor (PR). While ERalpha expression was not cycle-related, PR concentrations were high in the early and late-luteal phase and a regulatory role of both steroids on CL-function is assumed. As in other species also in the dog the immune system seems to participate in the mechanisms regulating CL-function as an increased presence of lymphocytes within the CL could be detected at the beginning [CD4- CD8-, major histocompatibility complex (MHC)II-antigen expressing cells] and during the latter half of dioestrus (CD8- and MHCII-antigen expressing cells). Thus, leucocyte-derived cytokines may be important and the expression of the mRNA for interleukin (IL)8, IL10, IL12, tumour necrosis factor (TNF)alpha and transforming growth factor (TGF) beta1 was observed throughout dioestrus. Electron microscopy confirmed the slow process of luteolysis; first distinct signs of degeneration were seen on day 60, accompanied by some apoptotic events. From these data it is concluded that luteal regression as monitored by the gradual decrease of systemic progesterone concentrations in the dog is not an actively regulated but rather a permissive process. Immune-mediated events may play a key role. Changes in the vascular supply, as indicated by the expression of endoglin, seem to be of lower importance.     

In vitro Evaluation of in vivo Fertilizing Ability of Frozen Rabbit Semen

The present work studied different spermatozoa parameters and the ability of frozen rabbit spermatozoa to fertilize, in vitro, in vivo‐matured oocytes, as a test to predict their in vivo fertility and prolificacy. Semen from rabbit bucks was frozen using two freezing protocols [in a freezer at ?30°C or in liquid nitrogen vapour (LNV)]. For the in vivo trial, females were inseminated with frozen‐thawed spermatozoa. Oocytes used for in vitro testing were recovered 14 h after ovulation induction from donors and co‐incubated with 2 × 10⁶ frozen‐thawed spermatozoa during 4 h at 37°C in Tyrode's medium under an atmosphere of 5% CO₂ in air with maximal humidity. After co‐incubation period, presumptive zygotes were cultured in TCM199 supplemented with 20% foetal bovine serum (FBS), under the same conditions described above. Although no statistical differences were observed between freezing protocols in seminal parameters [motility rate: 40 and 35%, VCL: 35 and 46 μm/s, amplitude of lateral head displacement (ALH): 1.7 and 2.4 μm, for semen frozen at ?30°C and in LNV, respectively], significant differences were noted in the fertilizing ability in vivo and in vitro. Semen frozen at ?30°C showed the highest fertilizing ability in vitro (26.7% vs 6.2 and 8.7% for semen frozen at ?30°C, in LNV and fresh semen, respectively) and the lowest fertility rate in vivo (21.7% vs 64.2% and 70.6% for semen frozen at ?30°C, in LNV and fresh semen, respectively). Sperm frozen at ?30°C seemed to be more capacitated.     

Association analyses between the prion protein locus and reproductive and lamb weight traits in Ripollesa sheep

The objective of this study was to analyze the association between the haplotypes of the prion protein (PrP) locus and several reproductive and lamb weight traits in Ripollesa sheep. Prion protein genotypes were available for a total of 310 sheep (7 rams, 114 ewes, and 189 lambs), all of them belonging to the purebred Ripollesa flock of the Universitat Autònoma of Barcelona, for which all sheep had a known pedigree. In addition, the genotype of 24 historical descendants of the previously genotyped adult individuals was reconstructed, provided that both parents were homozygous for PrP haplotypes. Only 3 haplotypes (ARR, ARQ, and ARH) were observed in the PrP locus of the sheep sampled. Reproductive traits included conception rate and litter size, whereas birth BW and 90-d BW were the lamb weight traits studied. The additive effect of PrP haplotypes was analyzed through Bayesian animal threshold and linear models, for reproduction and weight traits, respectively. Ewe reproductive data belonged to 89 ewes that gave 492 conception rate records and 440 litter size records. Analyses of BW at birth and at 90 d of age were made on 323 and 164 lamb records, respectively. No associations between PrP haplotypes and conception rate and BW traits were observed. For litter size, the effect of the ARH haplotype was greater than that of the ARQ haplotype. Differences between ARH and ARR haplotypes also suggested an advantage for the ARH. As a whole, our results indicated that the selection favorable to increase litter size in Ripollesa ewes may also increase the ARH haplotype frequency, which contradicts the recommendations of the current European Union legislation aiming to increase the genetic resistance to scrapie. As a consequence, scrapie genotyping needs to be included as a new selection criterion in the breed.