In tropical regions, protein supplementation is a common practice in dairy and beef farming. However, the effect of highly degradable protein in ruminal fermentation and microbial community composition has not yet been investigated in a systematic manner. In this work, we aimed to investigate the impact of casein supplementation on volatile fatty acids (VFA) production, specific activity of deamination (SAD), ammonia concentration and bacterial and archaeal community composition. The experimental design was a 4×4 Latin square balanced for residual effects, with four animals (average initial weight of 280±10 kg) and four experimental periods, each with duration of 29 days. The diet comprised Tifton 85 (Cynodon sp.) hay with an average CP content of 9.8%, on a dry matter basis. Animals received basal forage (control) or infusions of pure casein (230 g) administered direct into the rumen, abomasum or divided (50 : 50 ratio) in the rumen/abomasum. There was no differences (P>0.05) in ruminal pH and microbial protein concentration between supplemented v. non-supplemented animals. However, in steers receiving ruminal infusion of casein the SAD and ruminal ammonia concentration increased 33% and 76%, respectively, compared with the control. The total concentration of VFA increased (P<0.05) in steers receiving rumen infusion of casein. SAD and the microbial protein concentration did not vary significantly among treatments during the feeding cycle, but mean SAD values were greater in steers supplemented in the rumen and rumen/abomasum. Ruminal ammonia concentration was positively correlated with SAD in animals receiving ruminal infusion of casein. Polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE) analysis revealed low similarity between treatments, animals and time of sample collection. Richness analysis and determination of the Shannon–Wiener index indicated no differences (P>0.05) in species richness and diversity of γ-proteobacteria, firmicutes and archaea between non-supplemented Nellore steers and steers receiving casein supplementation in the rumen. However, species richness and the Shannon–Wiener index were lower (P<0.05) for the phylum bacteroidetes in steers supplemented with casein in the rumen compared with non-supplemented animals. Venn diagrams indicated that the number of unique bands varied considerably among individual animals and was usually higher in number for non-supplemented steers compared with supplemented animals. These results add new knowledge about the effects of ruminal and postruminal protein supplementation on metabolic activities of rumen microbes and the composition of bacterial and archaeal communities in the rumen of steers.

The objective of this study was to investigate the relationship between nitrogen (N) partitioning and isotopic fractionation in lactating goats consuming diets with a constant high concentration of N and increasing levels of water soluble carbohydrate (WSC). Eight lactating goats were offered four different ratios of WSC : N in the diet. A two-period incomplete cross-over design was used, with two goats assigned to each treatment in each period. N balance measurements were conducted, with measurement of feed N intake and total output of N in milk, faeces and urine. Treatment, period and infusion effects were tested using general ANOVA; the relationships between variables were analysed by linear regression. Dietary treatment and period had significant effects on dry matter (DM) intake (g/day). DM digestibility (g/kg DM) and N digestibility (g/kg N) increased as the ratio of WSC : N increased in the diet. No treatment effect was observed on milk urea N concentration (g/l) or urinary excretion of purine derivatives (mM/day). Although dietary treatment and period had significant effects on N intake, the change of N intake was small; no effect was observed for N partitioning among faeces, milk and urine. Milk, plasma and faeces were enriched in 15N compared with feed, whilst urine was depleted in 15N relative to feed. No significant relationship was established between N partitioning and isotopic fractionation. This study failed to confirm the potential to use N isotopic fractionation as an indicator of N partitioning in dairy goats when diets provided N in excess to requirements, most likely because the range of milk N output/N intake and urinary N output/N intake were narrow.

Mathematical models are important tools to estimate nutritional requirements and animal growth. Very few calf models generated from other countries with different feeding programs, environment and production systems have been evaluated. The objective of this paper is to evaluate two calf models: (i) the National Research Council (NRC) in 2001 and (ii) the updates published by Van Amburgh and Drackley in 2005 and inputted into Agricultural Modeling and Training Systems (AMTS, version 3.5.8). Data from 16 previous studies involving 51 diets for dairy calves under tropical conditions (n=485 calves, initial BW 37.5±4.35 kg and weaning weight of 62.0±10.16 kg) were used. The calves were fed with whole milk, milk replacer or fermented colostrum, plus starter (20.9±1.78% of CP). The accuracy of the average daily gain (ADG) prediction was evaluated by mean bias, mean square prediction error (MSPE), concordance correlation coefficient, bias correction factor (Cb), and regression between the observed and predicted values. The ADG observed from birth to weaning was 0.452±0.121 kg/day. Calves fed with whole milk had greater ADG compared with calves fed milk replacer (0.477 v. 0.379 kg/day) during the milk-feeding period. When all data were pooled (n=51 diets), predictions had a mean bias of −0.019 and 0.068 kg/day for energy-allowable gain using NRC and AMTS models, respectively. The regression equation between observed and predicted values obtained from energy of diets showed an intercept different from zero (P<0.0001) and slope that differed from unity (P<0.0001). In a second evaluation, when calves were fed only milk replacer, the energy-allowable gain from AMTS showed the lowest mean bias (0.008 kg/day) and 82.1% of the MSPE value originated from random errors. The lowest MSPE, the higher Cb value and no significant slope bias (P>0.05) indicate that the AMTS growth model resulted in accurate predictions for calves fed with milk replacer. However, within these latter two approaches, the goodness of fit (R2) was low, representing low precision. The weight gain estimated by the energy available from the diet was overestimated by 19 g/day when calculated by the NRC and underestimated by 68 g/day when calculated by AMTS. The reasons for this discrepancy need to be understood, for only then new models could be developed and parameterized to estimate animal performance in tropical conditions more accurately and precisely.

Diet influences animal body and tissue composition due to direct deposition and to the nutrients effects on metabolism. The influence of specific nutrients on the molecular regulation of lipogenesis is not well characterized and is known to be influenced by many factors including timing and physiological status. A trial was performed to study the effects of different dietary energy sources on lipogenic genes transcription in ham adipose tissue of Iberian pigs, at different growth periods and on feeding/fasting situations. A total of 27 Iberian male pigs of 28 kg BW were allocated to two separate groups and fed with different isocaloric feeding regimens: standard diet with carbohydrates as energy source (CH) or diet enriched with high oleic sunflower oil (HO). Ham subcutaneous adipose tissue was sampled by biopsy at growing (44 kg mean BW) and finishing (100 kg mean BW) periods. The first sampling was performed on fasted animals, while the last sampling was performed twice, with animals fasted overnight and 3 h after refeeding. Effects of diet, growth period and feeding/fasting status on gene expression were explored quantifying the expression of a panel of key genes implicated in lipogenesis and lipid metabolism processes. Quantitative PCR revealed several differentially expressed genes according to diet, with similar results at both timings: RXRG, LEP and FABP5 genes were upregulated in HO group while ME1, FASN, ACACA and ELOVL6 were upregulated in CH. The diet effect on ME1 gene expression was conditional on feeding/fasting status, with the higher ME1 gene expression in CH than HO groups, observed only in fasting samples. Results are compatible with a higher de novo endogenous synthesis of fatty acids (FA) in the carbohydrate-supplemented group and a higher FA transport in the oleic acid-supplemented group. Growth period significantly affected the expression of most of the studied genes, with all but PPARG showing higher expression in finishing pigs according to a pattern dissimilar from the usual in cosmopolitan pig breeds. Feeding/fasting status only influenced PPARG gene transcription. The lack of effects of feeding/fasting status on lipogenic gene expression and the higher ME1 response to diet in fasting samples than in postprandial sampling, suggest the persistence of de novo lipogenesis during fasting. Overall results improve the understanding of the influence of different factors on lipid metabolism regulation in Iberian pigs.

We have previously demonstrated that a sharp rise in feed intake (hyperphagia) and spontaneous liver steatosis could be experimentally induced in domestic Greylag geese by combining a short photoperiod and a sequence of feed restriction followed by ad libitum corn feeding during the fall and the winter. In this previous work, however, individual feed intake could not be recorded. The present study aimed at evaluating the relationship between level and pattern of hyperphagia and liver weight with an individual control of feed intake in individually housed (IH) geese, while comparing the performances with group housed (GH) geese. A total of 300 male geese of 19 weeks of age, were provided with corn ad libitum after an initial feed restriction period. From 21 to 23 weeks of age, the daylight duration was progressively reduced from 10 to 7 h and kept as such until the end of the experiment (week 36). In all, 30 GH and 30 IH geese were slaughtered at 19, 27, 30, 32 and 36 weeks of age. Feed intake was measured per group in GH geese and individually in IH geese. During the 1st week of corn feeding, the average feed intake rose up to 600 g/goose per day in GH geese but not in IH geese where the feed intake rose gradually from 300 to 400 g/day. The liver weight increased from 93 g (week 19) to 497 g (week 32; P<0.05) in GH birds. In IH birds, liver weights were, on average, much lower (ranging from 220 to 268 g) than in GH birds (P<0.05). In GH and IH bird, the variability in the individual response to corn feeding was very high (liver weight cv ranging from 63% to 83% depending on slaughter age). A close correlation between corn consumption and liver weight was evidenced in IH birds at each slaughter age (R2 ranging from 0.62 to 0.79), except at 36 weeks of age where this correlation was weak (R2=0.14). The variability in the extent of liver steatosis is very high and our results in IH birds clearly point out the major role of hyperphagia, mainly at the beginning of the ad libitum corn feeding period, on the development of spontaneous liver steatosis.

Plant flavonoids are generally regarded as natural replacers of synthetic growth promoters in poultry production. This study investigated the immunomodulatory effects of plant flavonoids, such as genistein and hesperidin, in lipopolysaccharide (LPS)-challenged broilers. A total of 700 21-day-old commercial Arbor Acres broiler chicks were randomly assigned into six treatment groups, each having six pens of 20 chicks/pen. Chicks were fed a basal diet without any additive (control, CON), 5 mg genistein/kg feed (G5), 20 mg hesperidin/kg (H20), or a basal diet with a combination of genistein and hesperidin (1 : 4) with doses of 5 mg/kg feed (GH5), 10 mg/kg (GH10) and 20 mg/kg (GH20) for 6 weeks. Half of the birds from each treatment were separated, and either challenged with 0·9% sodium chloride solution or Escherichia coli LPS (250 μg/kg BW) on days 16, 18 and 20. The results showed that both genistein and hesperidin improved (P<0.01) the plasma antioxidant status of growing broilers, by increasing total antioxidant capacity (TAOC), superoxide dismutase (SOD) activity and decreasing malondialdehyde production. LPS challenge further increased (P<0.05) TAOC and SOD levels. Regardless of LPS challenge, both genistein and hesperidin improved the humoral and mucosal immunity by increasing the intestinal intraepithelial lymphocyte numbers (P<0.01), as well as anti-Newcastle disease and anti-avian influenza antibody titers (P<0.05). Supplementation of both the plant flavonoids generally increased (P<0.05) the immune organs indices (spleen, thymus and bursa). Thus, supplementation of basal diet of broiler chicks, either with genistein or hesperidin, improved immune and antioxidant status of growing broilers. In addition, combined supplementation of both the flavonoids showed further improvement than individual compounds.

Increasing plant species diversity has been proposed as a means for enhancing annual pasture productivity and decreasing seasonal variability of pasture production facing more frequent drought scenarios due to climate change. Few studies have examined how botanical complexity of sown swards affects cow performance. A 2-year experiment was conducted to determine how sward botanical complexity, from a monoculture of ryegrass to multi-species swards (MSS) (grasses-legumes-forb), affect pasture chemical composition and nutritive value, pasture dry matter (DM) intake, milk production and milk solids production of grazing dairy cows. Five sward species: perennial ryegrass (L as Lolium), white clover and red clover (both referred to as T as Trifolium because they were always sown together), chicory (C as Cichorium) and tall fescue (F as Festuca) were assigned to four grazing treatments by combining one (L), three (LT), four (LTC) or five (LTCF) species. Hereafter, the LT swards are called mixed swards as a single combination of ryegrass and clovers, whereas LTC and LTCF swards are called MSS as a combination of at least four species from three botanical families. The experimental area (8.7 ha) was divided into four block replicates with a mineral nitrogen fertilisation of 75 kg N/ha per year for each treatment. In total, 13 grazing rotations were carried out by applying the same grazing calendar and the same pasture allowance of 19 kg DM/cow per day above 4 cm for all treatments. Clover represented 20% of DM for mixed and MSS swards; chicory represented 30% of DM for MSS and tall fescue represented 10% of DM for LTCF swards. Higher milk production (+1.1 kg/day) and milk solids production (+0.08 kg/day) were observed for mixed swards than for ryegrass swards. Pasture nutritive value and pasture DM intake were unaffected by the inclusion of clover. Pasture DM, organic matter and NDF concentrations were lower for MSS than for mixed swards. Higher milk production (+0.8 kg/day), milk solids production (+0.04 kg/day) and pasture DM intake (+1.5 kg DM/day) were observed for MSS than for mixed swards. These positive effects of MSS were observed for all seasons, but particularly during summer where chicory proportion was the highest. In conclusion, advantages of grazing MSS on cow performance were due to the cumulative effect of improved pasture nutritive value and increased pasture DM intake that raised milk production and milk solids production.

Two experiments were conducted to investigate the use of sorghum, cottonseed meal and millet in broiler diets and their interaction when they are used simultaneously. In Experiment 1, a corn-soybean meal control diet was compared with eight experimental treatments based on low tannin sorghum (S30, S45 and S60), cottonseed meal (CM15, CM40) or both ingredients included in the same diet (S30/CM40, S45/CM25 and S60CM15). Results showed that BW gain was not affected by the inclusion of sorghum or cottonseed meal. However, feed intake tended to be affected by the cereal type with the highest values with sorghum-based diets. Feed conversion ratio increased (P<0.001) with sorghum-based diets compared with the control diet, whereas a combination of cottonseed meal and sorghum in the same diet did not affect the feed conversion ratio. Significant differences (P<0.001) were observed in apparent ileal digestibility (%) of protein and energy with the cottonseed meal and sorghum/cottonseed meal-based diets having lower protein and energy digestibility compared with corn-based diets. In Experiment 2, a control diet was compared with six diets in which corn was substituted at 60%, 80% or 100% by either sorghum or millet and other three diets with simultaneous inclusion of these two ingredients (S30/M30, S40/M40, S50/M50). Single or combined inclusion of sorghum and millet resulted in similar feed intake and growth performance as the control diet. Apparent ileal digestibility of protein and energy was higher with millet-based diets (P<0.001). Total tract digestibility of protein in sorghum and millet-based diets tended to decrease linearly with the increasing level of substitution. Sorghum-based diets resulted in lower total tract digestibility of fat compared with millet and sorghum/millet-based diets (P<0.001). Higher total tract digestibility of starch were obtained with the control diet and millet-based diets compared with the sorghum-based treatments. Results of the two experiments suggest that broiler growth performance was not affected by the dietary level of sorghum, millet or cottonseed meal. Nutrient digestion can, however, be affected by these feed ingredients.

Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance and blood amino acid (AA) concentrations when fed to piglets. Experiment 1 included 18 pigs (15.0±1.1 kg) fed one of three diets containing either 0.63 or 0.69 standardized ileal digestible (SID) Val : Lys, or 0.63 SID Val : Lys supplemented with a Bacillus subtilis mutant (mutant 1). Blood samples were obtained 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding and analyzed for AAs. In Experiment 2, 80 piglets (9.1±1.1 kg) were fed one of four diets containing 0.63 or 0.67 SID Val : Lys, or 0.63 SID Val : Lys supplemented with another Bacillus subtilis mutant (mutant 2) or its parent wild type. Average daily feed intake, daily weight gain and feed conversion ratio were measured on days 7, 14 and 21. On day 17, blood samples were taken and analyzed for AAs. On days 24 to 26, six pigs from each dietary treatment were fitted with a permanent jugular vein catheter, and blood samples were taken for AA analysis 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding. In experiment 1, Bacillus subtilis mutant 1 tended (P<0.10) to increase the plasma levels of Val at 2 and 3 h post-feeding, but this was not confirmed in Experiment 2. In Experiment 2, Bacillus subtilis mutant 2 and the wild type did not result in a growth performance different from the negative and positive controls. In conclusion, results obtained with the mutant strains of Bacillus subtilis were not better than results obtained with the wild-type strain, and for both strains, the results were not different than the negative control.

Previously, the single nucleotide polymorphism in alcohol dehydrogenase (ADH1C c.-64T>C) was shown to have an association with intramuscular fat (IMF) in the longissimus thoracis (LT) muscle when vitamin A was limited in finishing rations of beef steers. The purpose of this study was to determine the optimum vitamin A supplementation level, in combination with ADH1C genotype, to increase IMF of the LT muscle. In total, 45 TT genotype, 45 CT and 27 CC Black Angus crossbred steers were backgrounded on a commercial ration containing 3360 IU vitamin A/kg dry matter (DM). During finishing, the steers were randomly assigned to one of three vitamin A treatments at 25%, 50% and 75% of the National Research Council recommendation of 2200 IU/kg DM. Treatments were administered via an oral bolus. Carcass quality was evaluated and a sample from the LT muscle was collected for analysis of IMF. A treatment×genotype interaction (P=0.04) was observed for IMF; TT steers on the 75% treatment had higher IMF relative to CT and CC steers on the same treatment. Western blot analysis showed that TT steers had higher (P=0.02) ADH1C protein expression in hepatic tissue. Previously, TT steers exhibited increased IMF when fed limited vitamin A. In the current study, the lack of variation in IMF between treatments and genotypes at the lower vitamin A treatment levels was likely due to the majority of the steers grading Canada AAA (USDA Choice). However, the western blot data supports that TT steers are expected to have higher IMF deposition, due to an increased production of ADH1C. The interaction between ADH1C genotype and vitamin A supplementation level has the potential for use in marker-assisted management programs to target niche markets based on increased marbling.

There is a global imperative to reduce phosphorous (P) excretion from pig systems. In this study, a previously validated deterministic model was modified to be stochastic, in order to investigate the consequences of different management strategies on P excretion by a group of growing pigs. The model predicts P digestion, retention and excretion from feed composition and growth parameters that describe a specified pig phenotype. Stochasticity was achieved by introducing random variation in the latter. The strategies investigated were: (1) changing feed composition frequently in order to match more closely pig digestible P (digP) requirements to feed composition (phase feeding) and (2) grouping pigs into light and heavy groups and feeding each group according to the requirements of their group average BW (sorting). Phase feeding reduced P excretion as the number of feeding phases increased. The effect was most pronounced as feeding phases increased from 1 to 2, with a 7.5% decrease achieved; the increase in phases from 2 to 3 was associated with a further 2.0% reduction. Similarly, the effect was more pronounced when the feed targeted the population requirements for digP at the average BW of the first third, rather than the average requirements at the mid-point BW of each feeding sequence plan. Increasing the number of feeding phases increased the percentage of pigs that met their digP requirements during the early stages of growth and reduced the percentage of pigs that were supplied <85% of their digP requirements at any stage of their growth; the latter may have welfare implications. Sorting of pigs reduced P excretion to a lesser extent; the reduction was greater as the percentage of pigs in the light group increased from 10% to 30% (from 1.5% to 3.0% reduction, respectively). This resulted from an increase in the P excreted by the light group, accompanied by a decrease in the P excreted by the remaining pigs. Sorting increased the percentage of light pigs that met their dig P requirements, but only slightly decreased the percentage of heavy pigs that met these requirements at any point of their growth. Exactly the converse was the case as far as the percentage of pigs that were supplied <85% of their digP requirements were concerned. The developed model is flexible and can be used to investigate the effectiveness of other management strategies in reducing P excretion from groups of pigs, including precision livestock feeding.

High-immune quality colostrum (IgG1 concentration ⩾50 g/l) is crucial for the health and development of the young calf. Studies on colostrum quality tend to focus on external factors such as breed, parity or dry period length, but few have focused on within-cow variations. Here we ran experiments to gain a deeper insight into within-cow variation in IgG1 concentrations in dairy cow colostrum. Trials were performed in an experimental farm, located in the Western part of France. Colostrum from each quarter and a composite sample (mix of four quarters) were concomitantly collected on 77 Holstein dairy cows just after calving to assess the influence of sample type on IgG1 concentrations. Variation in IgG1 concentrations during the first milking was studied on samples from nine cows collected every minute from the start of milking. Repeatability of colostral IgG1 concentration was estimated from 2009 and 2010 data on 16 healthy cows. IgG1 concentrations were tested using a radial immunodiffusion method. Sensitivity and specificity were similar regardless of sample type tested (individual quarter or composite milk). Mean average IgG1 concentration was 54.1 g/l in composite colostrum, and was significantly higher in hind quarter teats (56.2 g/l) than front quarter teats (53.1 g/l). Average IgG1 concentration did not change significantly during colostrum milking, and the variations observed (15% or less) were likely due to the laboratory method (CV 15%). IgG1 concentrations in dam colostrum increased slightly from 2009 to 2010 due to BW and parity effects. In 56% of cases, colostrum quality could have been assessed on either individual or composite colostrum samples collected at any time during the first milking without affecting the reliability of the measurement. However, in other cases, differences were significant enough to mean that estimates of average IgG1 concentration in colostrum from any one quarter would not be reliable. It is concluded that colostrum quality, from an IgG1 concentration point of view, could be assessed with a composite sample taken at any time during the first milking.

Reduction of the CP content in the diets of piglets requires supplementation with crystalline essential amino acids (AA). Data on the leucine (Leu) and histidine (His) requirements of young pigs fed low-CP diets are limited and have primarily been obtained from nonlinear models. However, these models do not consider the possible decline in appetite and growth that can occur when pigs are fed excessive amounts of AA such as Leu. Therefore, two dose-response studies were conducted to estimate the standardised ileal digestible (SID) Leu : lysine (Lys) and His : Lys required to optimise the growth performance of young pigs. In both studies, the average daily gain (ADG), average daily feed intake (ADFI) and gain-to-feed ratio (G : F) were determined during a 6-week period. To ensure that the diets had sub-limiting Lys levels, a preliminary Lys dose-response study was conducted. In the Leu study, 60 35-day-old piglets of both sexes were randomly assigned to one of five treatments and fed a low-CP diet (15%) with SID Leu : Lys levels of 83%, 94%, 104%, 115% or 125%. The His study used 120 31-day-old piglets of both sexes, which were allotted to one of five treatments and fed a low-CP diet (14%) with SID His : Lys levels of 22%, 26%, 30%, 34% or 38%. Linear broken-line, curvilinear-plateau and quadratic-function models were used for estimations of SID Leu : Lys and SID His : Lys. The minimum SID Leu : Lys level needed to maximise ADG, ADFI and G : F was, on average, 101% based on the linear broken-line and curvilinear-plateau models. Using the quadratic-function model, the minimum SID Leu : Lys level needed to maximise ADG, ADFI and G : F was 108%. Data obtained from the quadratic-function analysis further showed that a ±10% deviation from the identified Leu requirement was accompanied by a small decline in the ADG (−3%). The minimum SID His : Lys level needed to maximise ADG, ADFI and G : F was 27% and 28% using the linear broken-line and curvilinear-plateau models, respectively, and 33% using the quadratic-function model. The preferred model to estimate the His requirement was the curvilinear-plateau model. However, a 10% reduction in the SID His : Lys level was associated with an 11% reduction in the ADG. In conclusion, the SID Leu : Lys level needed to maximise growth was 108% when using the quadratic-function model as the best-fitting model. The minimum SID His : Lys level required to optimise growth was 28% when using the curvilinear-plateau model as the best-fitting model.

The ATP-binding cassette transporter G2/breast cancer resistance protein (ABCG2/BCRP) is an efflux protein involved in the bioavailability and milk secretion of endogenous and exogenous compounds, actively affecting milk composition. A limited number of physiological substrates have been identified. However, no studies have reported the specific effect of this polymorphism on the secretion into milk of compounds implicated in milk quality such as vitamins or endogenous compounds. The bovine ABCG2 Y581S polymorphism is described as a gain-of-function polymorphism that increases milk secretion and decreases plasma levels of its substrates. This work aims to study the impact of Y581S polymorphism on plasma disposition and milk secretion of compounds such as riboflavin (vitamin B2), enterolactone, a microbiota-derived metabolite from the dietary lignan secoisolariciresinol and uric acid. In vitro transport of these compounds was assessed in MDCK-II cells overexpressing the bovine ABCG2 (WT-bABCG2) and its Y581S variant (Y581S-bABCG2). Plasma and milk levels were obtained from Y/Y homozygous and Y/S heterozygous cows. The results show that riboflavin was more efficiently transported in vitro by the Y581S variant, although no differences were noted in vivo. Both uric acid and enterolactone were substrates in vitro of the bovine ABCG2 variants and were actively secreted into milk with a two-fold increase in the milk/plasma ratio for Y/S with respect to Y/Y cows. The in vitro ABCG2-mediated transport of the drug mitoxantrone, as a model substrate, was inhibited by enterolactone in both variants, suggesting the possible in vivo use of this enterolignan to reduce ABCG2-mediated milk drug transfer in cows. The Y581S variant was inhibited to a lesser extent probably due to its higher transport capacity. All these findings point to a significant role of the ABCG2 Y581S polymorphism in the milk disposition of enterolactone and the endogenous molecules riboflavin and uric acid, which could affect both milk quality and functionality.

Replacing dairy components from milk replacer (MR) with vegetable products has been previously associated with decreased protein and fat digestibility in milk-fed calves resulting in lower live weight gain. In this experiment, the major carbohydrate source in MR, lactose, was partly replaced with gelatinized corn starch (GCS) to determine the effect on protein and fat digestibility in milk-fed calves. In total, 16 male Holstein-Friesian calves received either MR with lactose as the carbohydrate source (control) or 18% GCS at the expense of lactose. In the adaptation period, calves were exposed to an increasing dose of GCS for 14 weeks. The indigestible marker cobalt ethylenediaminetetraacetic acid was incorporated into the MR for calculating apparent nutrient digestibility, whereas a pulse dose of chromium (Cr) chloride was fed with the last MR meal 4 h before slaughter as an indicator of passage rates. The calves were anesthetized and exsanguinated at 30 weeks of age. The small intestine was divided in three; small intestine 1 and 2 (SI1 and SI2, respectively) and the terminal ileum (last ~100 cm of small intestine) and samples of digesta were collected. Small intestinal digesta was analysed for α-amylase, lipase and trypsin activity. Digestibility of protein was determined for SI1, SI2, ileum and total tract, whereas digestibility of fat was determined for SI1, SI2 and total tract. Apparent protein digestibility in the small intestine did not differ between treatments but was higher in control calves at total tract level. Apparent crude fat digestibility tended to be increased in SI1 and SI2 for GCS calves, but no difference was found at total tract level. Activity of α-amylase in SI2 and lipase in both SI1 and SI2 was higher in GCS calves. Activity of trypsin tended to be higher in control calves and was higher in SI1 compared with SI2. A lower recovery of Cr in SI2 and a higher recovery of Cr in the large intestine suggest an increased rate of passage for GCS calves. Including 18% of GCS in a milk replacer at the expense of lactose increased passage rate and decreased apparent total tract protein digestibility. In the small intestine, protein digestion did not decrease when feeding GCS and fat digestion even tended to increase. Overall, effects on digestion might be levelled when partially replacing lactose with GCS, because starch digestion is lower than that of lactose but fat digestion may be slightly increased when feeding GCS.

European legislation states that after stunning regular checks should be performed to guarantee animals are unconscious between the end of the stunning process and death. When animals are killed without prior stunning these checks should be performed before the animal is released from restraint. The validity of certain indicators used to assess unconsciousness under different stunning and slaughter conditions is under debate. The aim of this study was to validate the absence of threat-, withdrawal-, corneal- and eyelid reflex as indicators to assess unconsciousness in calves subjected to different stunning and slaughter methods. Calves (201±22 kg) were randomly assigned to one of the following four treatments: (1) Captive bolt stunning followed by neck cut in an inverted position (n=25); (2) Non-stunned slaughter in an upright position (n=7); (3) Non-stunned slaughter in an inverted position (180° rotation) (n=25); (4) Non-stunned slaughter in an upright position followed by captive bolt stunning 40 s after the neck cut (n=25). Each calf was equipped with non-invasive electroencephalogram (EEG) electrodes before the slaughter procedure. All reflexes were verified once before the slaughter procedure. At the beginning of the procedure (T=0 s) calves were stunned (treatment 1) or neck cut in an upright position (treatment 2, 4) or inverted position (treatment 3). Calves of treatment 4 were captive bolt stunned 34±8 s after the neck cut. Reflexes were assessed every 20 s from T=15 s for all treatments until all reflex tests resulted in a negative response three times in a row and a flat line EEG was observed. In addition, reflexes were assessed 5 s after captive bolt stunning in calves of treatments 1 and 4. Visual assessment of changes in the amplitude and frequency of EEG traces was used to determine loss of consciousness. Timing of loss of consciousness was related to timing of loss of reflexes. After captive bolt stunning, absence of threat-, withdrawal-, corneal- and eyelid reflex indicated unconsciousness as determined by EEG recordings. After non-stunned slaughter, both threat- and withdrawal reflex were on average lost before calves were unconscious based on EEG recordings. The eyelid- and corneal reflex were on average lost after calves had lost consciousness based on EEG recordings and appeared to be distinctly conservative indicators of unconsciousness in non-stunned slaughtered calves since they were observed until 76±50 and 85±45 s (mean±SD), respectively, after EEG-based loss of consciousness.

Adding nitrate to the diet or increasing the concentration of dietary lipid are effective strategies for reducing enteric methane emissions. This study investigated their effect on health and performance of finishing beef cattle. The experiment was a two×two×three factorial design comprising two breeds (CHX, crossbred Charolais; LU, Luing); two basal diets consisting of (g/kg dry matter (DM), forage to concentrate ratios) 520 : 480 (Mixed) or 84 : 916 (Concentrate); and three treatments: (i) control with rapeseed meal as the main protein source replaced with either (ii) calcium nitrate (18 g nitrate/kg diet DM) or (iii) rapeseed cake (RSC, increasing acid hydrolysed ether extract from 25 to 48 g/kg diet DM). Steers (n=84) were allocated to each of the six basal diet×treatments in equal numbers of each breed with feed offered ad libitum. Blood methaemoglobin (MetHb) concentrations (marker for nitrate poisoning) were monitored throughout the study in steers receiving nitrate. After dietary adaptation over 28 days, individual animal intake, performance and feed efficiency were recorded for a test period of 56 days. Blood MetHb concentrations were low and similar up to 14 g nitrate/kg diet DM but increased when nitrate increased to 18 g nitrate/kg diet DM (P<0.001). An interaction between basal diet and day (P<0.001) indicated that MetHb% was consistently greater in Concentrate – than Mixed-fed steers at 18 g nitrate/kg diet DM. Maximum individual MetHb% was 15.4% (of total Hb), which is lower than considered clinically significant (30%). MetHb concentrations for individual steers remained consistent across time. Concentrate-fed steers were more efficient (lower residual feed intake (RFI) values) than Mixed-fed steers (P<0.01), with lower dry matter intake (DMI) (kg/day) (P<0.001) and similar average daily gain (ADG). CHX steers were more efficient (lower RFI; P<0.01) than LU steers with greater ADG (P<0.01), lower DMI (/kg BW; P<0.01) and lower fat depth (P<0.001). ADG, BW or DMI did not differ across dietary treatments (P>0.05). Neither basal diet nor treatment affected carcass quality (P>0.05), but CHX steers achieved a greater killing out proportion (P<0.001) than LU steers. Thus, adding nitrate to the diet or increasing the level of dietary lipid through the use of cold-pressed RSC, did not adversely affect health or performance of finishing beef steers when used within the diets studied.

Two experiments were conducted to evaluate the effects of a novel synthetic emulsifier product (AVI-MUL TOP) on the growth performance of chickens for fattening and weaned piglets. The emulsifier product consists of 50% vegetal bi-distillated oleic acid emulsified with 50% glyceryl polyethyleneglycol ricinoleate. In experiment 1, 480 1-day-old female Cobb500 chickens for fattening were assigned to two treatments: (1) a control diet (CTR); and (2) the control diet+the emulsifier (AMT, 1 g/kg from day 0 to day 10, 0.75 g/kg from day 10 to day 20 and 0.5 g/kg from day 20 to day 34 of the trial). AMT supplementation increased BW on days 20 and 34 (P<0.01). Dietary AMT increased the average daily gain and average daily feed intake (ADFI) from day 10 to day 20, from day 20 to day 34 and from day 0 to day 34 (P<0.01). A reduced feed conversion ratio was observed in the AMT group from day 10 to day 20 (P<0.01). In experiment 2, 96 Stambo HBI×Dalland piglets were weaned at 24 days and assigned to two treatments (the basal diet without the product (CTR) or with 2 g/kg emulsifier from day 0 to day 14 and 1.5 g/kg from day 14 to day 42 (AMT)). There was an increase in the ADFI associated with AMT supplementation from day 14 to day 42 (P=0.04). These results indicated that supplementation with the synthetic emulsifier may significantly improve the growth performance of chickens for fattening and numerically improve that of weaned piglets.

Physically effective fiber is needed by dairy cattle to prevent ruminal acidosis. This study aimed to examine the effects of different sources of physically effective fiber on the populations of fibrolytic bacteria and methanogens. Five ruminally cannulated Holstein cows were each fed five diets differing in physically effective fiber sources over 15 weeks (21 days/period) in a Latin Square design: (1) 44.1% corn silage, (2) 34.0% corn silage plus 11.5% alfalfa hay, (3) 34.0% corn silage plus 5.1% wheat straw, (4) 36.1% corn silage plus 10.1% wheat straw, and (5) 34.0% corn silage plus 5.5% corn stover. The impact of the physically effective fiber sources on total bacteria and archaea were examined using denaturing gradient gel electrophoresis. Specific real-time PCR assays were used to quantify total bacteria, total archaea, the genus Butyrivibrio, Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens and three uncultured rumen bacteria that were identified from adhering ruminal fractions in a previous study. No significant differences were observed among the different sources of physical effective fiber with respect to the microbial populations quantified. Any of the physically effective fiber sources may be fed to dairy cattle without negative impact on the ruminal microbial community.

It is imperative to evaluate precise nutrient requirements of animals in order to optimize productivity and minimize feed cost and nutrient excretions. The current non-phytate phosphorus (NPP) recommendation for broilers is based on the papers published 30 years ago. However, today’s commercial birds are quite different from those before 30 years. Therefore, the present experiment was conducted with growing male broiler chickens to evaluate an optimal dietary NPP level of broiler chickens fed a conventional corn–soybean meal diet from 4 to 6 weeks of age. The 1-day-old chicks were fed corn–soybean meal diet containing 0.39% NPP from 1 to 3 weeks of age. At 22 days of age, 360 birds were selected and randomly allotted by BW to one of 10 dietary treatments with six replicate cages of six birds per cage for each treatment. Birds were fed the P-unsupplemented corn–soybean meal basal diet and the basal diet supplemented with inorganic P as CaHPO4·H2O ranging from 0.00% to 0.45% with 0.05% increment from 4 to 6 weeks of age. The dietary NPP levels were 0.09%, 0.14%, 0.20%, 0.24%, 0.30%, 0.34%, 0.38%, 0.45%, 0.49% and 0.54%, respectively, and the dietary Ca level was fixed at 0.90% for all treatments. The results showed that average daily gain, serum inorganic P concentration, tibia bone strength, tibia ash percentage and P percentage, tibia bone mineral content (BMC) and density (BMD), middle toe ash percentage and P percentage, middle toe BMC, total body BMC and BMD were affected (P<0.0001) by dietary NPP level, and increased linearly (P<0.0001) and quadratically (P<0.003) as dietary NPP levels increased. Optimal dietary NPP levels estimated based on fitted broken-line models (P<0.0001) of the above indices are 0.21%, 0.29%, 0.29%, 0.29%, 0.29%, 0.31%, 0.29%, 0.30%, 0.27%, 0.29% and 0.28%, respectively. It is suggested that the total body BMC and BMD, and middle toe ash P and BMC might be new, sensitive and non-invasive criteria to evaluate the dietary NPP requirements of broilers. The optimal dietary NPP level would be 0.31% for broiler chickens fed a conventional corn–soybean meal diet from 4 to 6 weeks of age.