The objective of this study was to determine the effects of pistachio by-product silage (PBPS) as a partial replacement for corn silage (CS) on chewing activity, nutrients digestibility and ruminal fermentation parameters in Holstein male calves over a 6-month assay. For this purpose, 24 Holstein male calves (4 to 5 months of age and 155.6±13.5 kg BW) were randomly assigned to one of the four dietary treatments (n=6). In these treatments, CS was substituted with different levels of PBPS (0%, 6%, 12% and 18% of dry matter (DM)). Nutrient digestibility was measured at the end of the experimental period (days 168 to 170). Ruminal fermentation parameters were determined on days 90 and 180 and chewing activity was determined on days 15 of the 3rd and 6th month of the experiment. Results showed that calves fed rations containing 6% PBPS spent more time ruminating (P<0.05) than the control group on the 3rd and 6th months. Feeding PBPS was found to have no effects on DM, organic matter (OM), ether extract or ash digestibility, but apparent digestibility of CP, NDFom and ADFom linearly decreased (P<0.01) with increasing substitutions. On days 90 and 180, ruminal concentrations of volatile fatty acids and NH3–N linearly decreased (P<0.01) with increasing levels of PBPS in the diets; however, ruminal pH and molar proportions of acetate, propionate and butyrate were similar across the treatments. It was concluded that partial substitution of CS with PBPS (6% or 12%) would have no adverse effects on nutrient digestibility, total chewing activity and ruminal fermentation parameters.

Glucocorticoid receptor (GR) transcription is driven by alternative promoters to produce different exon 1 mRNA variants. CpG methylation on GR promoters profoundly affects GR transcription. GR in hippocampus is critical for energy homeostasis and stress responses, yet it remains unclear whether hippocampal expression of GR exon 1 mRNA variants and the methylation status of GR promoters differ between Large White (LW) and Erhualian (EHL) pigs showing distinct metabolic and stress-coping characteristics. EHL pigs had higher hippocampus weight relative to BW (P<0.01), which was associated with higher serum cortisol level compared with LW pigs. Hippocampal expression of brain-derived neurotrophic factor (P<0.05) was significantly higher, while Bax, a pro-apoptotic gene, was significantly lower in EHL pigs (P<0.05). Hippocampal expression of total GR did not differ between breeds, yet GR exon 1 to 11 mRNA was significantly higher (P<0.01) in EHL pigs, which was associated with a trend of increase (P=0.057) in GR protein content. No significant breed difference was detected for the methylation status across the whole region of the proximal GR promoter, while CpG334 and CpG266.267 were differentially methylated, in a reversed manner, between breeds. The methylation status of CpGs 248, 259, 260, 268 and 271 was negatively correlated (P<0.05) with GR exon 1 to 11 mRNA abundance. Our results provide fundamental information on the breed-specific characteristics of GR and its mRNA variants expression and the status of DNA methylation on the proximal GR promoter in the pig hippocampus.

This study aimed to identify genetic evaluation models (GEM) to accurately select cattle for milk production when only limited data are available. It is based on a data set from the Pakistani Sahiwal progeny testing programme which includes records from five government herds, each consisting of 100 to 350 animals, with lactation records dating back to 1968. Different types of GEM were compared, namely: (1) multivariate v. repeatability model when using the first three lactations, (2) an animal v. a sire model, (3) different fixed effects models to account for effects such as herd, year and season; and (4) fitting a model with genetic parameters fixed v. estimating the genetic parameters as part of the model fitting process. Two methods were used for the comparison of models. The first method used simulated data based on the Pakistani progeny testing system and compared estimated breeding values with true breeding values. The second method used cross-validation to determine the best model in subsets of actual Australian herd-recorded data. Subsets were chosen to reflect the Pakistani data in terms of herd size and number of herds. Based on the simulation and the cross-validation method, the multivariate animal model using fixed genetic parameters was generally the superior GEM, but problems arise in determining suitable values for fixing the parameters. Using mean square error of prediction, the best fixed effects structure could not be conclusively determined. The simulation method indicated the simplest fixed effects structure to be superior whereas in contrast, the cross-validation method on actual data concluded that the most complex one was the best. In conclusion it is difficult to propose a universally best GEM that can be used in any data set of this size. However, some general recommendations are that it is more appropriate to estimate the genetic parameters when evaluating for selection purposes, the animal model was superior to the sire model and that in the Pakistani situation the repeatability model is more suitable than a multivariate.

Twenty ruminally cannulated beef heifers were fed a high corn grain diet in a randomized block design to determine the effect of three direct fed microbial (DFM) strains of Propionibacterium on ruminal fermentation, nutrient digestibility and methane (CH4) emissions. The heifers were blocked in five groups on the basis of BW and used in five 28-day periods. Dietary treatments included (1) Control and three strains of Propionibacterium (2) P169, (3) P5, and (4) P54. Strains were administered directly into the rumen at 5×109 CFU with 10 g of a maltodextrin carrier in a gel capsule; Control heifers received carrier only. All heifers were fed the basal diet (10 : 90 forage to concentrate, dry matter basis). Rumen contents were collected on days 15 and 18, ruminal pH was measured continuously between days 15 and 22, enteric CH4 emissions were measured between days 19 and 22 and diet digestibility was measured from days 25 to 28. Mean ruminal pH was 5.91 and was not affected by treatments. Similarly, duration of time that pH<5.8 and 5.6 was not affected by treatment. Likewise, total and major volatile fatty acid profiles were similar among all treatments. No effects were observed on dry matter intake and total tract digestibility of nutrients. Total enteric CH4 production (g/day) was not affected by Propionibacterium strains and averaged 139 g/day. Similarly, mean CH4 yield (g CH4/kg of dry matter intake) was similar for all the treatments. The relative abundance of total Propionibacteria in the rumen increased with administration of DFM and were greater 3 h post-dosing relative to Control, but returned to baseline levels before feeding. Populations of Propionibacterium P169 were higher at 3 and 9 h as compared with the levels at 0 h. In conclusion, moderate persistency of the inoculated strains within the ruminal microbiome and pre-existing high propionate production due to elevated levels of starch fermentation might have reduced the efficacy of Propionibacterium strains to increase molar proportion of propionate and subsequently reduce CH4 emissions.

Glutamate, which is one of the most important contributors to oxidative metabolism in the intestinal mucosa, is mainly transported by the excitatory amino acids transporters (EAATs) that are expressed in enterocytes. The objective of this study was to evaluate the effects of in ovo administration of l-trans pyrrolidine-2,4-dicarboxylic acid (l-trans-PDC), a potent competitive inhibitor of glutamate uptake by EAATs, on the growth of the small intestine in chicks. Two series of experiments were conducted with hatching eggs; 100 μl of various l-trans-PDC solutions (0, 0.075 or 0.225 mg/egg for the Control group, low-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (L-PDC) or high-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (H-PDC), respectively) was injected into the albumen sac of these hatching eggs before incubation. Hatchlings were sacrificed by cervical dislocation to determine the embryonic development in Experiment I, whereas the birds in Experiment II were raised or sampled at hatching, days 7 and 14 (D7 and D14) for further study. Gene expression in the small intestines was determined by real-time RT-PCR; and serum concentration of free amino acids was determined by an amino acid analyzer. The results showed that the hatchability was decreased by in ovo administration of l-trans-PDC. The small intestinal weights of the H-PDC group were decreased (P<0.05) at hatching and increased (P<0.05) on D7 and D14 compared with those in the Control group. In addition, the gene expression of EAAT2 in the completed or segmental small intestines was not changed (P>0.05); EAAT3 gene expression in the duodenum (P<0.05), jejunum (P=0.084) and ileum (P=0.060) on D14 was lower in the H-PDC group than in the Control group. Furthermore, the serum concentrations of free proline, threonine and phenylalanine but not glutamate or aspartate were increased (P<0.06) in H-PDC group. In conclusion, this paper is the first to report that in ovo administration of l-trans-PDC induces small intestinal growth retardation during the embryonic period and catch-up growth after hatching.

In milk-fed calves, quantification of the milk that enters the rumen (ruminal milk volume, RMV) because of malfunction of the esophageal groove reflex may explain part of the variability observed between animals in their growth performance. The RMV can directly be quantified by adding an indigestible marker to the diet and measuring its recovery in the rumen at slaughter, but this technique cannot be repeated in time in the same animal. The objective of the study was to evaluate three indirect methods for estimating RMV. The first method was based on the assumption that ruminal drinking delays and limits acetaminophen appearance in blood after ingestion of milk supplemented with acetaminophen. The second method was based on a negative linear relationship between RMV and urinary recovery of non-metabolizable monosaccharides (3-O-methylglucose, l-rhamnose and d-xylose) added to the milk, owing to rumen fermentation. In the third method, RMV was calculated as the difference between total milk intake and the increase in abomasal milk volume (AMV) at feeding, measured through ultrasonography shortly after feeding, or estimated from the mathematical extrapolation of AMV to feeding time, based on consecutive measurements. These methods were tested in three experiments where calves (n=22, 10 and 13) were bucket fed or partly tube fed (i.e. by inserting milk replacer into the rumen via a tube to mimic ruminal drinking). In addition, Co-EDTA and Cr-EDTA were used as an indigestible marker in one experiment to trace bucket-fed or tube-fed milk replacer, respectively, to measure RMV. The relationship between AMV measured by ultrasonography and AMV measured at slaughter improved when kinetics of AMV were extrapolated to the time of slaughter by mathematical modeling (error between predicted and measured AMV equaled 0.49 l). With this technique, RMV during feeding averaged 17% and 24% of intake in Experiments 2 and 3, respectively. Plasma acetaminophen kinetics and recovery of non-metabolizable monosaccharides in urine were partly associated with ruminal drinking, but these techniques are not considered quantitatively accurate without further information of rumen degradation and absorption. The recovery of indigestible marker measured at slaughter gave a quantitative estimate of RMV (2% in Experiment 3), but improper measurement of emptying rate of fluid from the rumen may lead to underestimation. In conclusion, measuring changes in AMV by ultrasonography, in response to milk feeding, was the most promising indirect method to quantify RMV in veal calves.

This study aimed to investigate the effects and possible interactions of birth weight and n-3 polyunsaturated fatty acid (PUFA) supplementation of the maternal diet on the fatty acid status of different tissues of newborn piglets. These effects are of interest as both parameters have been associated with pre-weaning mortality. Sows were fed a palm oil diet or a diet containing 1% linseed, echium or fish oil from day 73 of gestation. As fish oil becomes a scarce resource, linseed and echium oil were supplemented as sustainable alternatives, adding precursor fatty acids for DHA to the diet. At birth, the lightest and heaviest male piglet per litter were killed and samples from liver, brain and muscle were taken for fatty acid analysis. Piglets that died pre-weaning had lower birth weights than piglets surviving lactation (1.27±0.04 v. 1.55±0.02 kg; P<0.001), but no effect of diet on mortality was found. Lower DHA concentrations were observed in the brain of the lighter piglets compared with their heavier littermates (9.46±0.05 v. 9.63±0.04 g DHA/100 g fatty acids; P=0.008), suggesting that the higher incidence of pre-weaning mortality in low birth weight piglets may be related to their lower brain DHA status. Adding n-3 PUFA to the sow diet could not significantly reduce this difference in DHA status, although numerically the difference in the brain DHA concentration between the piglet weight groups was smaller when fish oil was included in the sow diet. Independent of birth weight, echium or linseed oil in the sow diet increased the DHA concentration of the piglet tissues to the same extent, but the concentrations were not as high as when fish oil was fed.

Next-generation sequencing (NGS; also known as deep sequencing or ultra-high throughput sequencing) has probably been the most important tool for genomic research over the past few years. NGS has led to numerous discoveries and scientific breakthroughs in the genetic field. The sequencing technology that has entered the research laboratory in the past decade is now being introduced into the clinical diagnostic laboratory. Consequently, NGS results are becoming available in the medical arena as abundance of clinically relevant variants, conferring predisposition to disease, are being discovered at a growing rate (Stanley, 2014).

The study aimed to determine the effects of red clover (Trifolium pratense) silage with high phyto-oestrogen content on ewe performance during their first breeding season. Red clover silage containing formononetin, biochanin A, genistein, and daidzein was fed to 10 nulliparous ewes of the prolific Finnish Landrace breed before, during and after the breeding season, for a total of 5 months. A control group of 10 ewes was fed with grass silage. The mean numbers of foetuses per pregnancy were 2.1±0.7 and 2.2±0.8 for the red clover and control groups, respectively. The total mass of the uterus with its contents was significantly greater in ewes of the red clover group compared with those of the control group. This difference was mainly explained by the greater volume of foetal fluids. Serum progesterone concentration in the red clover group was significantly lower over the entire period analysed than in the control group. In conclusion, the fecundity of the ewes was not reduced by red clover feed with high phyto-oestrogen concentrations. The volume of foetal fluids increased that could increase the risk for vaginal prolapse before the term.

Dairy cow mortality is an important animal welfare issue that also causes financial losses. The objective of this study was to identify farm characteristics and herd management practices associated with high on-farm cow mortality in Swedish dairy herds. A postal questionnaire was sent to farmers that had either high or low mortality rates for 3 consecutive years. The questionnaire consisted of five sections: ‘About the farm’, ‘Milking and housing’, ‘Feeding’, ‘Routines’ and ‘Lame and sick cows’. A total of 145 questionnaires were returned (response rate=33%). Ten of the 77 characteristics investigated met the inclusion criteria for multivariable analysis. The final logistic regression model included: herd size, breed, use of natural service bull, bedding improvement frequency and pasture system. Herds with Swedish Holstein as the predominant breed (odds ratio (OR) 22.9, 95% confidence interval (CI) 5.2 to 101.8) or with mixed breeds (OR 5.5, 95% CI 1.7 to 17.5) had a greater risk of being high mortality herds than herds that were predominantly Swedish Red (OR 1). Herds larger than 100 cows (OR 19.6, 95% CI 3.5 to 110.4) and herds with 50 to 99 cows (OR 13.8, 95% CI 3.2 to 60.6) had greater risk of mortality than herds numbering 35 to 50 cows (OR 1). Being a high mortality herd was also associated with having cows on exercise lots during the summer season (OR 3.6, 95% CI 1.3 to 9.9) compared with on pasture. A missing answer on the question of bedding improvement frequency was associated with high mortality herds. Overall, this study suggests that characteristics that are related to intensification of the dairy industry are also associated with high on-farm mortality of dairy cows.

The most common housing system for reproduction rabbits, individual cage housing on a wire floor, is increasingly scrutinized because of its potential detrimental impact on animal welfare. We compared three types of housing: (1) individual cage housing on a wire floor (3952 cm2/doe, maximum roof height 63 cm, one 1000 cm2 plastic footrest/doe), (2) semi-group housing on a wire floor (5000 cm2/doe, roofless, one 1000 cm2 plastic footrest/doe) and (3) the same semi-group housing, but with a fully plastic slatted floor. In all housing systems, does had free access to an elevated platform. In the semi-group housing pens, four does were housed communally during 21 days of the reproduction cycle (to allow more space for locomotion and to increase opportunities for social contact), and individually during the other 21 days of the cycle (to minimize doe–doe and doe–kit aggression that peaks around kindling). In all, 24 Hycole does were included per system. The does entered the experiment at 203 days of age (after their first parity). The experiment consisted of four reproductive cycles, ending at 369 days of age. Pododermatitis was scored in cycles 1, 2 and 4. At the end of the 4th cycle the does were euthanized and X-rays were taken to assess spinal deformation. Tibia and femur length, width and cortical thickness were determined and bone strength was assessed using a shear test, as a measure of bone quality. Although severe pododermatitis was absent, the prevalence of plantar hyperkeratosis (hair loss and callus formation) at the end of the 4th cycle was much greater on the wire floor (65% and 68% for semi-group housing and individual cages, respectively) than on the plastic floor (5%, P<0.0001), even though the wire floors were equipped with a plastic footrest known to decrease hyperkeratosis. In contrast to our expectations, semi-group housing did not affect the prevalence of spinal deformations (P>0.10), but in line with our expectations bone quality was affected favourably by semi-group housing. The tibial cortex (and to a lesser extent the femoral cortex) was thicker in semi-group housing than in individual cages (1.45, 1.46 and 1.38 mm for semi-group housing on wire, semi-group housing on plastic and individual housing on wire, respectively, P=0.045). What this increase in cortical thickness means in terms of doe welfare requires further study, as it may reflect an increase in activity resulting either from increased space for locomotion, or from fleeing aggressive pen mates.

The effects of castration age, dietary protein level and the dietary lysine/methionine (lys/met) ratio on animal performance, carcass characteristics and meat quality were studied in 64 intensively reared Friesian steers. Animals underwent castration procedures at 15 days old or at 5 months old. Dietary treatments started at 90 days old, with eight animals from each castration age randomly allocated to each treatment: 14.6% v. 16.8% CP (DM basis), and 3.0 v. 3.4 lys/met, on a 2×2×2 design. The recommended ratio of 3.0 was reached with supplementation of protected methionine. Steers were slaughtered at 443.5±26.2 kg live weight when they reached 12 months old approximately. Average daily gain, cold carcass weight or carcass classification were not affected by any studied effect. Muscle moisture (P=0.024), C18:2n-6 percentage (P=0.047), polyunsaturated fatty acid/saturated fatty acid (P=0.049) and n-6/n-3 (P=0.003) were higher in late castrated animals. Both high levels of dietary protein (P=0.008) and lys/met ratio (P=0.048) increased the percentage of muscle in the carcass. A level of 16.8% of CP in the diet also increased the percentage of monounsaturated fatty acids in the intramuscular fat (P=0.032), whereas a ratio lys/met of 3.4 decreased the percentage of saturated fatty acids (P=0.028). Thus, it is recommended using diets with a high protein level (16.8%) and a high lys/met ratio (3.4) in animals slaughtered at a young age, in order to obtain carcasses with high muscle content without negatively affecting productive traits or intramuscular fat composition.