Risk factors for uterine diseases on small- and medium-sized dairy farms determined by clinical,bacteriological, and cytological examinations

The involution process of the postpartum bovine uterus is usually accompanied by invasion of various bacteria. The objectives of this study were to identify the relationship between the postpartum findings as risk factors for clinical endometritis (CE) and subclinical endometritis (SE). Furthermore, the effects of CE or SE on reproductive performance in small- and medium-sized dairy herds were investigated. A total of 400 cows were examined by vaginoscopy for CE at 20 to 30 days postpartum, and samples were collected for cytological examinations for SE and for bacteriology by cytobrush technique. The vaginoscopic and cytological examinations showed that 27.3% and 21.0% of the cows were found with CE and SE, respectively. The bacterial community analyses revealed a large variety of bacteria. Overall, bacteria from the order Actinomycetales, Lactobacillales, Bacillales, Burkholderiales, Caulobacteriales Enterobacteriales, Pasteurellales, and Pseudomonadales were detected, whereas in 39.5% of the samples no bacterial growth was detectable. The uterine pathogens Escherichia coli and Trueperella pyogenes were found in 16.8% and 13.0% of the samples cultivated under aerobic conditions. Other frequently isolated bacteria were Streptococcus spp. (31.3%), Staphylococcus spp. (20.0%), Corynebacterium spp. (16.5%), and Bacillus spp. (10.5%). The infection with T. pyogenes was the most important bacteriological risk factor for the occurrence of CE (odds ratio (OR) = 5.72; 95% CI = 3.07–10.83) and had a detrimental effect on the hazard of nonpregnancy by 200 days postpartum (hazard ratio = 1.66; 95% CI = 1.12–2.46). Calving assistance (OR = 1.79; 95% CI = 1.16–2.98) and farm (OR = 1.11; 95% CI = 1.02–1.20) were indicated as further risk factors for CE and SE. Effects of CE and SE on reproductive performance parameters could not be demonstrated.     

Characterization of DGAT1 Allelic Effects in a Sample of North American Holstein Cattle

A putative causative mutation underlying a QTL was identified as a lysine to alanine non-conservative substitution at amino acid 232 of the gene encoding the acylCoA:diacylglycerol acyltransferase (DGAT1) protein. Our goal was to characterize the allelic substitution effects of this DGAT1 mutation in a large sample of Holstein bulls from North America. Statistically significant effects were identified for all of the milk production traits and somatic cell scores. Estimated average effects of substituting the lysine allele for the alanine variant on Holstein bull daughter yield deviations were ?81 kg, 3.7 kg, ?1.1 kg, 0.063%, 0.012%, and ?0.023 units for milk yield, fat yield, protein yield, fat component, protein component, and SCS, respectively. These estimates were largely in agreement with previous studies; however, the magnitudes of the estimates were much smaller in this study. Impacts on economic indices for net merit, cheese merit, and fluid merit were modest. Because of the strong antagonism between fat and protein yield and how those traits influence economic indices, selection for DGAT1 genotypes will likely not find widespread application in the U.S.     

Effects of two levels of monoterpene blend on rumen fermentation, terpene and nutrient flows in the duodenum and milk production in dairy goats

Twenty-six Alpine and Saanen goats, 12 fitted with a rumen cannula and a T-type cannula in the duodenum, were used in a 6-week experiment on the effect of a monoterpene blend on rumen fermentation, duodenal terpene and nutrient flows, milk yield and composition. The monoterpene blend consisted of 45.2, 36.7, 16.0 and 2.2 mol/100 mol terpene for linalool, p-cymene, α-pinene and β-pinene, respectively. The four compounds were considered as good models of northwest Mediterranean sward terpenes and were supplied in the same proportions as in the spring diet of dairy goats in the Basilicata (Southern Apennine range, Italy). The goats were assigned to three experimental groups fed a total mixed ration with no terpene or with the monoterpene blend at two levels, 0.043 and 0.43 g/kg dry matter intake. Ruminal fermentation characteristics remained unchanged among groups. In average, the total volatile fatty acids (VFA) concentration was 89.0 mM and the proportions of acetate, propionate and butyrate were 66.4, 21.1 and 10.1 mol/100 mol VFA, with an acetate:propionate ratio of 3.19. In average, 0.0, 2.7, 3.5 and 23.5 mol/100 mol of the ingested amounts of linalool, α-pinene, p-cymene and β-pinene were recovered in the duodenum. The mean terpene composition of duodenal digesta was 11.7, 9.2 and 79.1 mol/100 mol terpene for p-cymene, α-pinene and β-pinene, differing markedly from the dietary blend in both supplemented groups. Apparent rumen digestibility of dry matter (0.40), of neutral detergent fibre (0.54), and of acid detergent fibre (0.45) was not affected by either level of terpene supplementation. Dry matter intake and milk production, milk fat content or milk fatty acid profile were not affected by terpenes. It is concluded that, because of extensive ruminal degradation of each terpene, the monoterpene blend had no effect on ruminal digestion of dietary constituents or on milk performance whatever the supplementation level.     

Detection of early pregnancy-specific proteins in Holstein milk

Bovine pregnancy is commonly diagnosed by rectal palpation or ultrasonography and changes in progesterone concentration. To determine a simpler and less expensive diagnostic method, we sought to identify early pregnancy-specific proteins in bovine milk by comparing samples collected from pregnant and non-pregnant Holstein cattle. Of the 600-700 protein spots visible on 2-DE gel images, 39 were differentially expressed in milk from pregnant and non-pregnant cattle. Antibodies generated against synthetic peptides of milk whey proteins expressed specifically during pregnancy were used to confirm protein expression patterns. Western blot analysis showed that the levels of expression of lactoferrin (lactotransferrin) and alpha1G T-type calcium channel subunit (alpha-1G) were higher in samples from pregnant than non-pregnant cattle. These findings suggest that assays for pregnancy-specific milk proteins may be used to diagnose pregnancy in cattle.     

Glutathione S-transferase Mu 3 is associated to in vivo fertility,but not sperm quality,in bovine

In the dairy breeding industry, pregnancy of dairy cows is essential to initiate milk production, so that high fertility rates are required to increase their productivity. In this regard, sperm proteins that are indicative of sperm quality and/or fertility have become an important target of study. Glutathione S-transferase Mu 3 (GSTM3) has been established as a fertility and sperm quality parameter in humans and pigs and, consequently, it might be a potential biomarker in cattle. For this reason, the present work aimed to determine if GSTM3 could predict sperm quality and in vivo fertility in this species. Sperm quality was assessed with flow cytometry and computer-assisted sperm analysis. Immunoblotting and immunofluorescence analysis were performed to determine the presence and localisation pattern of sperm GSTM3. This enzyme was found to be present in bovine sperm and to be localised along the sperm tail and the equatorial segment of the head. No significant associations between sperm GSTM3 and sperm quality parameters were observed, except a negative association with morphologically abnormal sperm having a coiled tail. In addition, and more relevant, higher levels of GSTM3 in sperm were seen in bulls showing lower in vivo fertility rates. In conclusion, our data evidenced the presence of GSTM3 in bovine sperm. Moreover, we suggest that, despite not being associated with sperm quality, GSTM3 might be an in vivo subfertility biomarker in cattle sperm, and that high levels of this protein could be an indicative of defective spermatogenesis and/or epididymal maturation.     

Estimation of cold stress effect on dairy cows

Twelve crossbred heifers (Slovak Spotted x Holstein-Friesian) were housed in an open, uninsulated barn with straw bedding and a concrete-floored yard. Minimum temperatures inside the barn were as low as –19°C. The average milk yield decreased as the temperatures approached these minima. Compared with the temperate conditions, the feed intake and blood levels of glucose and free fatty acids increased. The level of sodium declined significantly during the second cold period. Correlations and regressions between milk yield and biochemical parameters were calculated, and the results indicate that the concentrations of free fatty acids, cholesterol, and triiodothyronine and the haematocrit values may serve to predict milk production during periods of cold stress, or in lactations of 305 days.     

The preservation of liposomes by raffinose family oligosaccharides during drying is mediated by effects on fusion and lipid phase transitions

Raffinose family oligosaccharides (RFO) have been implicated as protective agents in the cellular dehydration tolerance, especially of many plant seeds. However, their efficacy in stabilizing membranes during dehydration has never been systematically investigated. We have analyzed the effects of sucrose, raffinose, stachyose, and verbascose on liposome stability during air-drying. With increasing degree of polymerization (DP), the RFO were progressively better able to stabilize liposomes against leakage of aqueous content and against membrane fusion after rehydration. Indeed, there was a very tight linear correlation between fusion and leakage for all RFO. These data indicate that increased protection of liposomes against leakage with increasing DP is due to better protection against fusion. This is in accord with the higher glass transition temperature of the longer chain oligosaccharides. Further evidence for the influence of glass transitions on membrane stability in the dry state was provided by experiments testing the temperature dependence of membrane fusion. During incubation at temperatures up to 95 °C for 2 h, fusion increased less with temperature in the presence of higher DP sugars. This indicates that RFO with a higher glass transition temperature are better able to protect dry membranes at elevated temperatures. In addition, Fourier-transform infrared (FTIR) spectroscopy showed a reduction of the gel to liquid-crystalline phase transition temperature of dry liposomes in the presence of all investigated sugars. However, the RFO became slightly less effective with increasing chain length, again pointing to a decisive role for preventing fusion. A direct interaction of the RFO with the lipids was indicated by a strong effect of the sugars on the phosphate asymmetric stretch region of the infrared spectrum.     

Effects of a ruminally protected B-vitamin supplement on milk yield and composition of lactating dairy cows

It is not clear if B vitamins supplied to the small intestine of dairy cows from dietary and rumen microbial sources are provided in sufficient quantity to maximize animal performance. Our objective was to determine effects of adding a ruminally protected B vitamin blend supplement, containing biotin, folic acid, pantothenic acid and pyridoxine, to the diet of high producing dairy cows on their productivity. Two dairy facilities located in California (USA) were used, one with mid lactation Holstein cows (Experiment 1) and the other with early lactation Holstein cows (Experiment 2). In each Experiment, cows were randomly assigned to treatment in a 2 × 2 crossover design with 28 d (Experiment 1) or 35 d (Experiment 2) experimental periods. In Experiment 1, milk and milk fat yield were unaffected by treatment, although milk fat proportion was lower (37.1 versus 36.3 g/kg; P<0.01), but milk protein yield was higher (1.21 versus 1.24 kg/d; P=0.02) in cows fed B vitamins. In Experiment 2, milk (39.60 versus 40.46 kg/d; P=0.02), milk fat (1.40 versus 1.47 kg/d; P<0.01) and milk protein yield (1.10 versus 1.16 kg/d; P<0.01), as well as milk energy output (113.2 versus 117.8 MJ/d; P<0.01) were all higher with B vitamin feeding. Body condition score (BCS) increased more with B vitamin feeding in Experiment 2, but was unaffected in Experiment 1. Body locomotion score (BLS) increased with B vitamin feeding in both experiments (P=0.01 and < 0.01, respectively), possibly an indication of reduced locomotory ability. Overall, productivity of high producing lactating dairy cows responded positively to feeding a mixture of ruminally protected B vitamins, although differences in the extent of the positive responses between experiments perhaps suggests that early lactation cows, with lower DM intake to milk yield ratios, may be more responsive to ruminally protected B vitamins than mid lactation cows, with higher DM intake to milk yield ratios.     

Mink Growth Hormone Structural–Functional Relationships: Effects of Renaturing and Storage Conditions

Fourier-transform infrared spectroscopy, in vitro bioassay and enzyme-linked immunoassay were used to study the structural-functional relationships of recombinant mink growth hormone (mGH), refolded and stored under different conditions. Porcine GH (pGH) was synthesized and used as an example. These two hormones, when refolded and stored the same way, had the same secondary structures, biological and immunological efficacy, and biological potency. Only the immunological potency differed, mGH being significantly less potent than pGH. Renaturation pH and storing frozen or at 4 °C in 5% glycerol did not affect either the secondary structure or the activity. However, freeze-drying raised the content of buried α-helices and lowered that of solvated α-helices and of unordered structures. These conformational changes were associated with a reduction of immunological and biological potency of mGH and of immunological potency of pGH. These findings provide original information on the secondary structure of mGH, and show that conformational changes induced by lyophilization adversely affect its activity.