Breeding and genetics
Research Article
Multi-generational imputation of single nucleotide polymorphism marker genotypes and accuracy of genomic selection
- S. Toghiani, S. E. Aggrey, R. Rekaya
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- Published online by Cambridge University Press:
- 06 January 2016, pp. 1077-1085
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Availability of high-density single nucleotide polymorphism (SNP) genotyping platforms provided unprecedented opportunities to enhance breeding programmes in livestock, poultry and plant species, and to better understand the genetic basis of complex traits. Using this genomic information, genomic breeding values (GEBVs), which are more accurate than conventional breeding values. The superiority of genomic selection is possible only when high-density SNP panels are used to track genes and QTLs affecting the trait. Unfortunately, even with the continuous decrease in genotyping costs, only a small fraction of the population has been genotyped with these high-density panels. It is often the case that a larger portion of the population is genotyped with low-density and low-cost SNP panels and then imputed to a higher density. Accuracy of SNP genotype imputation tends to be high when minimum requirements are met. Nevertheless, a certain rate of genotype imputation errors is unavoidable. Thus, it is reasonable to assume that the accuracy of GEBVs will be affected by imputation errors; especially, their cumulative effects over time. To evaluate the impact of multi-generational selection on the accuracy of SNP genotypes imputation and the reliability of resulting GEBVs, a simulation was carried out under varying updating of the reference population, distance between the reference and testing sets, and the approach used for the estimation of GEBVs. Using fixed reference populations, imputation accuracy decayed by about 0.5% per generation. In fact, after 25 generations, the accuracy was only 7% lower than the first generation. When the reference population was updated by either 1% or 5% of the top animals in the previous generations, decay of imputation accuracy was substantially reduced. These results indicate that low-density panels are useful, especially when the generational interval between reference and testing population is small. As the generational interval increases, the imputation accuracies decay, although not at an alarming rate. In absence of updating of the reference population, accuracy of GEBVs decays substantially in one or two generations at the rate of 20% to 25% per generation. When the reference population is updated by 1% or 5% every generation, the decay in accuracy was 8% to 11% after seven generations using true and imputed genotypes. These results indicate that imputed genotypes provide a viable alternative, even after several generations, as long the reference and training populations are appropriately updated to reflect the genetic change in the population.
Crossbreeding effects on rabbit reproduction from four maternal lines of rabbits
- M. Ragab, J. P. Sánchez, C. Mínguez, M. Baselga
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- 07 January 2016, pp. 1086-1092
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Litter size is essential for an efficient production of rabbit meat. A diallel cross between four maternal lines was carried out and the analysis of the components of litter size has been already done. This paper presents the analysis of litter size traits themselves (total born (TB), number born alive (NBA), number weaned (NW)) and kindling interval (KI), that complete the analysis of the reproductive performance. The 16 genetic groups were distributed in four Spanish farms. The V line was present in all farms in order to be used as reference group. A total of 34 546 parities from 7111 does, were analysed. The crossbreeding parameters were estimated according to Dickerson model. The differences between lines performance were of low magnitude and not significant for litter size traits. The LP line showed the shortest KI followed by H respect to lines A and V. These differences reflected the differences between direct and maternal genetic effects. The differences between the average of all crosses and line V were found to be significant and seemed to be important, being 0.46 for TB, 0.56 for NBA, 0.75 for NW and −2.21 days for KI. The differences between reciprocal crosses for litter size were of low magnitude and non-significant, which indicate that the maternal effects are not important between these lines. In general, the lines did not show significant differences in direct and maternal genetic effects for TB, NBA and NW but there were some significant differences for KI, which ranged from 1.54 to 6.85 days in direct effects and from 0.63 to 3.38 days for maternal effects. A positive and, in some cases, relevant heterosis was found. The largest heterosis was for TB in the HV cross (1.05 rabbits), followed by the AH (0.74 rabbits), AV (0.57 rabbits) and LH (0.55 rabbits) crosses. For NBA, significant heterosis was found in HV (1.11 rabbits) and AV (0.49 rabbits) and for NW in AV (0.90 rabbits), LH (0.70 rabbits) and LV (0.58 rabbits). Favourable and significant heterosis for KI was found in AV and LV crosses, whereas it was unfavourable in AL and in LH. The more recommended crosses were AV and LH, showing the greatest performances on NW, while the cross HV, which shows the greatest performances an NBA, could be highly recommended when cross-fostering is practised to equalise litter sizes after birth, in this situation the lower NW performance observed could be improved.
Multiple-breed reaction norm animal model accounting for robustness and heteroskedastic in a Nelore–Angus crossed population
- M. M. Oliveira, M. L. Santana, F. F. Cardoso
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- 12 January 2016, pp. 1093-1100
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Our objective was to genetically characterize post-weaning weight gain (PWG), over a 345-day period after weaning, of Brangus-Ibagé (Nelore×Angus) cattle. Records (n=4016) were from the foundation herd of the Embrapa South Livestock Center. A Bayesian approach was used to assess genotype by environment (G×E) interaction and to identify a suitable model for the estimation of genetic parameters and use in genetic evaluation. A robust and heteroscedastic reaction norm multiple-breed animal model was proposed. The model accounted for heterogeneity of residual variance associated with effects of breed, heterozygosity, sex and contemporary group; and was robust with respect to outliers. Additive genetic effects were modeled for the intercept and slope of a reaction norm to changes in the environmental gradient. Inference was based on Monte Carlo Markov Chain of 110 000 cycles, after 10 000 cycles of burn-in. Bayesian model choice criteria indicated the proposed model was superior to simpler sub-models that did not account for G×E interaction, multiple-breed structure, robustness and heteroscedasticity. We conclude that, for the Brangus-Ibagé population, these factors should be jointly accounted for in genetic evaluation of PWG. Heritability estimates increased proportionally with improvement in the environmental conditions gradient. Therefore, an increased proportion of differences in performance among animals were explained by genetic factors rather than environmental factors as rearing conditions improved. As a consequence response to selection may be increased in favorable environments.
Nutrition
Research Article
Rabbit dietary supplementation with pale purple coneflower. 1. Effects on the reproductive performance and immune parameters of does
- S. Dabbou, L. Rotolo, A. Kovitvadhi, S. Bergagna, D. Dezzutto, R. Barbero, P. Rubiolo, A. Schiavone, M. De Marco, A. N. Helal, I. Zoccarato, L. Gasco
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- 14 January 2016, pp. 1101-1109
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Echinacea pallida (EPAL), also known as pale purple coneflower, is a herbaceous flowering plant with immune-enhancement and antioxidative properties. The effect of EPAL on the reproductive performance, serum biochemistry and haematological parameters of rabbit does has been studied here. A total of 100, 21-week-old Grimaud rabbit does, were randomly assigned to two groups. One group was fed a basal diet supplemented with 3 g EPAL/kg diet (Echinacea group, E), while the other was fed the basal diet without the supplementation (control group, C). The reproductive performance of the does was not affected by the treatment (P>0.05). The haematological parameters of pregnant rabbits showed that there was no interaction between gestation day and treatment. The EPAL supplementation induced a reduction (−47.3%) in the basophil cell rate (0.55% and 0.29%, for the control and treatment groups, respectively; P=0.049). The gestation day significantly affected most of the haematological parameters (P<0.05). The white blood cell counts declined progressively after day 14. The mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red cell distribution width, mean platelet volume and eosinophils increased steadily throughout the study, and reached a maximum value on day 28. The red blood cells, haemoglobin, haematocrit, mean corpuscular volume and neutrophils increased slightly up to day 14, and then subsequently decreased progressively until day 28. The lymphocytes and platelet distribution width decreased until day 14, and then increased to a maximum value on day 28. No significant effect of gestation day or treatment was observed on the blood serum chemistry. As far as the immune parameters are concerned, no significant differences were observed between groups, while a significant effect of gestation day was observed for lysozymes (6.02 v. 7.99 v. 1.91; for 0, 14 and 28 days, respectively; P=0.014). In conclusion, a lack of effect of EPAL has been observed. In fact, no impacts of EPAL have been observed on the reproductive or haematological parameters of the does. The effects of dietary supplementation with EPAL on the performances, bacterial community, blood parameters and immunity in growing rabbits are reported in the second part of this study.
Rabbit dietary supplementation with pale purple coneflower. 2. Effects on the performances, bacterial community, blood parameters and immunity of growing rabbits
- A. Kovitvadhi, F. Gai, S. Dabbou, I. Ferrocino, L. Rotolo, M. Falzone, C. Vignolini, M. S. Gennero, S. Bergagna, Daniela Dezzutto, R. Barbero, P. Nebbia, S. Rosati, L. Cocolin, I. Zoccarato, L. Gasco
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- 14 January 2016, pp. 1110-1117
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Echinacea pallida (EPAL), a herbaceous flowering plant with immunomodulatory properties, has been chosen to determine the pre- and post-supplementary effects on the growth performances, bacterial community, blood parameters and immunity of growing rabbits. The same Grimaud does (14-week-old) from the studied in the first part of this study were randomly divided into two groups (n=50/group). The first group was fed a basal diet without supplementation (Control group, C) while the another group was fed a basal diet supplemented with 3 g EPAL/kg diet (Echinacea group, E). From the second parturition, 80 weaned kits (40 from the C does and 40 from the E does) were randomly assigned to four groups of 20 animals each and were fed a growing commercial diet supplemented with or without a 3 g EPAL/kg diet: the CC group (rabbits from the C does fed the control diet), CE group (rabbits from the C does fed the supplemented diet), EC (rabbits from the E does fed the control diet) and EE group (rabbits from the E does fed the supplemented diet). The dietary EPAL treatment did not affect the growth performance. Ten fattening rabbits from each group were selected to evaluate the bacterial community and blood parameters, while the remaining rabbits (n=10/group) were used to study phagocytosis and the humoral immune response. The variability was evaluated from hard faeces at 35, 49 and 89 days, and the caecal content at 89 days. The variability of the bacterial community of the EE group was higher than that of the other groups. The phagocytic activity was higher in the CE and EE groups than in the CC and EC ones (30.9 and 29.7 v. 21.2 and 21.8%; P<0.05), whereas no statistically significant difference was observed for the blood parameters or humoral immune response against vaccination (rabbit haemorrhagic disease virus) at 95 days old which the serum was collected at 88, 102, 109, 116 and 123 days old. In conclusion, no impact of EPAL dietary supplementation has been observed on the growth performances, bacterial community, blood parameters or humoral immune responses in growing rabbits, except for an increase in phagocytic activities.
Simultaneous inclusion of sorghum and cottonseed meal or millet in broiler diets: effects on performance and nutrient digestibility
- D. I. Batonon-Alavo, D. Bastianelli, P. Lescoat, G. M. Weber, M. Umar Faruk
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- Published online by Cambridge University Press:
- 03 February 2016, pp. 1118-1128
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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.
Dietary supplementation of pyrroloquinoline quinone disodium protects against oxidative stress and liver damage in laying hens fed an oxidized sunflower oil-added diet
- J. Wang, H. J. Zhang, L. Xu, C. Long, K. G. Samuel, H. Y. Yue, L. L. Sun, S. G. Wu, G. H. Qi
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- 03 February 2016, pp. 1129-1136
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The protective effects of dietary pyrroloquinoline quinone disodium (PQQ.Na2) supplementation against oxidized sunflower oil-induced oxidative stress and liver injury in laying hens were examined. Three hundred and sixty 53-week-old Hy-Line Gray laying hens were randomly allocated into one of the five dietary treatments. The treatments included: (1) a diet containing 2% fresh sunflower oil; (2) a diet containing 2% thermally oxidized sunflower oil; (3) an oxidized sunflower oil diet with 100 mg/kg of added vitamin E; (4) an oxidized sunflower oil diet with 0.08 mg/kg of PQQ.Na2; and (5) an oxidized sunflower oil diet with 0.12 mg/kg of PQQ.Na2. Birds fed the oxidized sunflower oil diet showed a lower feed intake compared to birds fed the fresh oil diet or oxidized oil diet supplemented with vitamin E (P=0.009). Exposure to oxidized sunflower oil increased plasma malondialdehyde (P<0.001), hepatic reactive oxygen species (P<0.05) and carbonyl group levels (P<0.001), but decreased plasma glutathione levels (P=0.006) in laying hens. These unfavorable changes induced by the oxidized sunflower oil diet were modulated by dietary vitamin E or PQQ.Na2 supplementation to levels comparable to the fresh oil group. Dietary supplementation with PQQ.Na2 or vitamin E increased the activities of total superoxide dismutase and glutathione peroxidase in plasma and the liver, when compared with the oxidized sunflower oil group (P<0.05). PQQ.Na2 or vitamin E diminished the oxidized sunflower oil diet induced elevation of liver weight (P=0.026), liver to BW ratio (P=0.001) and plasma activities of alanine aminotransferase (P=0.001) and aspartate aminotransferase (P<0.001) and maintained these indices at the similar levels to the fresh oil diet. Furthermore, oxidized sunflower oil increased hepatic DNA tail length (P<0.05) and tail moment (P<0.05) compared with the fresh oil group. Dietary supplementation of PQQ.Na2 or vitamin E decreased the oxidized oil diet induced DNA tail length and tail moment to the basal levels in fresh oil diet. These results indicate that PQQ.Na2 is a potential antioxidant and is as effective against oxidized oil-related liver injury in laying hens as vitamin E. The protective effects of PQQ.Na2 against liver damage induced by oxidized oil may be partially due to its role in the scavenging of free radicals, inhibiting of lipid peroxidation and enhancing of antioxidant defense systems.
Precision feeding can significantly reduce lysine intake and nitrogen excretion without compromising the performance of growing pigs
- I. Andretta, C. Pomar, J. Rivest, J. Pomar, J. Radünz
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- 13 January 2016, pp. 1137-1147
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This study was developed to assess the impact on performance, nutrient balance, serum parameters and feeding costs resulting from the switching of conventional to precision-feeding programs for growing–finishing pigs. A total of 70 pigs (30.4±2.2 kg BW) were used in a performance trial (84 days). The five treatments used in this experiment were a three-phase group-feeding program (control) obtained with fixed blending proportions of feeds A (high nutrient density) and B (low nutrient density); against four individual daily-phase feeding programs in which the blending proportions of feeds A and B were updated daily to meet 110%, 100%, 90% or 80% of the lysine requirements estimated using a mathematical model. Feed intake was recorded automatically by a computerized device in the feeders, and the pigs were weighed weekly during the project. Body composition traits were estimated by scanning with an ultrasound device and densitometer every 28 days. Nitrogen and phosphorus excretions were calculated by the difference between retention (obtained from densitometer measurements) and intake. Feeding costs were assessed using 2013 ingredient cost data. Feed intake, feed efficiency, back fat thickness, body fat mass and serum contents of total protein and phosphorus were similar among treatments. Feeding pigs in a daily-basis program providing 110%, 100% or 90% of the estimated individual lysine requirements also did not influence BW, body protein mass, weight gain and nitrogen retention in comparison with the animals in the group-feeding program. However, feeding pigs individually with diets tailored to match 100% of nutrient requirements made it possible to reduce (P<0.05) digestible lysine intake by 26%, estimated nitrogen excretion by 30% and feeding costs by US$7.60/pig (−10%) relative to group feeding. Precision feeding is an effective approach to make pig production more sustainable without compromising growth performance.
Ileal digestibility of nutrients and amino acids in low quality soybean meal sources treated with β-mannanase for growing pigs
- S. D. Upadhaya, J. W. Park, J. H. Lee, I. H. Kim
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- 09 February 2016, pp. 1148-1154
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Apparent ileal digestibility (AID) of energy, dry matter (DM), nitrogen (N) and amino acids and standardized ileal digestibility (SID) of amino acids in low quality soybean meals with different CP concentration (SBM 44% CP and SBM 48% CP) with or without 400 U β-mannanase/kg supplementation were evaluated in 20 cannulated barrows ((Landrace×Yorkshire)×Duroc) with an average BW of 25.08±3.42 kg. A N-free diet was used to determine basal endogenous losses of amino acids. The supplementation of β-mannanase improved (P<0.05) AID of DM compared with non-supplemented diet. However, enzyme supplementation did not improve (P>0.05) AID of N and energy. The type of SBM (SBM 44% CP v. SBM 48% CP) had no effect on AID of DM, N and energy. β-mannanase improved (P<0.05) AID of sum of essential amino acids, arginine, histidine, lysine, valine and glycine. The SID of lysine was higher (P<0.05) in enzyme supplemented than in non-supplemented diets. Larger AID and SID of threonine and proline (P<0.05) were observed in SBM 48% CP than in SBM 44% CP. In conclusion, the supplementation of enzyme improved AID of arginine, histidine, lysine, valine and glycine, but it did not cause marked difference in SID of these amino acids except for lysine. The low nutrient digestibility of the SBM sources used in the present experiment might have favoured the positive effect of β-mannanase supplementation.
Metabolic response to dietary fibre composition in horses
- C. Brøkner, D. Austbø, J. A. Næsset, D. Blache, K. E. Bach Knudsen, A. H. Tauson
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- 12 January 2016, pp. 1155-1163
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The hypothesis for this study was that a higher dietary proportion of soluble fibre would result in stable and constant plasma metabolite and regulatory hormone concentrations. The study was a 4×4 Latin Square design with a sequence of 17 days adaptation to the ration followed by 8 sampling days. The feed rations consisted of only timothy hay (H), hay plus molassed sugar beet pulp combined with either whole oats (OB) or barley (BB) and hay plus a loose chaff-based concentrate (M). Four horses were fitted with permanent caecal cannulas and liquid caecal content was withdrawn manually and blood was drawn from the jugular vein at 0, 3 and 9 h postprandial. The horses were exercised daily at medium level for about 1 h. Samples were analysed for short-chain fatty acids (SCFA) and metabolic traits. Caecal SCFA and propionic acid concentrations increased with increased dietary starch and soluble fibre. The diet highest in soluble fibre (M) resulted in the highest plasma glucose and insulin concentrations in the morning, which then remained stable and constant throughout the day. A strong interaction (P<0.01) between time and diet was measured for plasma urea, glucose, insulin and leptin. The greatest variations in plasma glycaemic and insulinaemic responses were associated with the cereal grain diets (OB and BB). There were indications of a negative energy balance, which was reflected in a significantly higher plasma β-hydroxybutyrate concentration and a numerically higher non-esterified fatty acid concentration. In conclusion, this study found that inclusion of soluble fibre resulted in increased total caecal SCFA and propionic acid concentrations. This consequently resulted in stable and constant plasma glycaemic and insulinaemic responses. Diets with a high content of soluble fibre provided enough energy for horses at medium work level.
Relative contribution of ruminal buffering systems to pH regulation in feedlot cattle fed either low- or high-forage diets
- G. E. Chibisa, K. A. Beauchemin, G. B. Penner
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- 06 January 2016, pp. 1164-1172
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The relative contribution of ruminal short-chain fatty acid (SCFA) absorption and salivary buffering to pH regulation could potentially change under different dietary conditions. Therefore, the objective of this study was to investigate the effects of altering the ruminal supply of rapidly fermentable carbohydrate (CHO) on absorptive function and salivation in beef cattle. Eight heifers (mean BW±SD=410±14 kg) were randomly allocated to two treatments in a crossover design with 37-day periods. Dietary treatments were barley silage at 30% low forage (LF) or 70% high forage (HF) of dietary dry matter (DM), with the remainder of the diet consisting of barley grain (65% or 25% on a DM basis) and a constant level (5%) of supplement. The LF and HF diets contained 45.3% and 30.9% starch, and 4.1% and 14.0% physically effective fiber (DM basis), respectively. Ruminal pH was continuously measured from day 17 to day 23, whereas ruminal fluid was collected on day 23 to determine SCFA concentration. Ruminal liquid passage rate was determined on day 23 using Cr-ethylenediaminetetraacetic acid. Eating or resting salivation was measured by collecting masticate (days 28 and 29) or saliva samples (days 30 and 31) at the cardia, respectively. On days 30 and 31, the temporarily isolated and washed reticulo-rumen technique was used to measure total, and Cl−-competitive (an indirect measure of protein-mediated transport) absorption of acetate, propionate and butyrate. As a result of the higher dietary starch content and DM intake, the ruminal supply of rapidly fermentable CHO, total ruminal SCFA concentration (118 v. 95 mM; P<0.001) and osmolality (330 v. 306 mOsm/kg; P=0.018) were greater in cattle fed LF compared with HF. In addition, feeding LF resulted in a longer duration (2.50 v. 0.09 h/day; P=0.02) and a larger area (0.44 v. 0.01 (pH×h)/day; P=0.050) that pH was below 5.5. There was no diet effect on total and Cl−-competitive absorption (mmol/h and %/h) of acetate, propionate, butyrate and total SCFA (acetate+propionate+butyrate), but eating salivation was less (131 v. 152 ml/min; P=0.02), and resting salivation tended to be less (87 v. 104 ml/min; P=0.10) in cattle fed an LF diet. In summary, lower ruminal pH in cattle with greater rapidly fermentable CHO intake was attributed to an increase in SCFA production and decrease in salivation, which were not compensated for by an increase in epithelial permeability.
Long-term effect of linseed plus nitrate fed to dairy cows on enteric methane emission and nitrate and nitrite residuals in milk
- J. Guyader, M. Doreau, D. P. Morgavi, C. Gérard, C. Loncke, C. Martin
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- 06 January 2016, pp. 1173-1181
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A previous study showed the additive methane (CH4)-mitigating effect of nitrate and linseed fed to non-lactating cows. Before practical application, the use of this new strategy in dairy cows requires further investigation in terms of persistency of methanogenesis reduction and absence of residuals in milk products. The objective of this experiment was to study the long-term effect of linseed plus nitrate on enteric CH4 emission and performance in dairy cows. We also assessed the effect of this feeding strategy on the presence of nitrate residuals in milk products, total tract digestibility, nitrogen (N) balance and rumen fermentation. A total of 16 lactating Holstein cows were allocated to two groups in a randomised design conducted in parallel for 17 weeks. Diets were on a dry matter (DM) basis: (1) control (54% maize silage, 6% hay and 40% concentrate; CON) or (2) control plus 3.5% added fat from linseed and 1.8% nitrate (LIN+NIT). Diets were equivalent in terms of CP (16%), starch (28%) and NDF (33%), and were offered twice daily. Cows were fed ad libitum, except during weeks 5, 16 and 17 in which feed was restricted to 95% of dry matter intake (DMI) to ensure complete consumption of meals during measurement periods. Milk production and DMI were measured weekly. Nitrate and nitrite concentrations in milk and milk products were determined monthly. Daily CH4 emission was quantified in open circuit respiration chambers (weeks 5 and 16). Total tract apparent digestibility, N balance and rumen fermentation parameters were determined in week 17. Daily DMI tended to be lower with LIN+NIT from week 4 to 16 (−5.1 kg/day on average). The LIN+NIT diet decreased milk production during 6 non-consecutive weeks (−2.5 kg/day on average). Nitrate or nitrite residuals were not detected in milk and associated products. The LIN+NIT diet reduced CH4 emission to a similar extent at the beginning and end of the trial (−47%, g/day; −30%, g/kg DMI; −33%, g/kg fat- and protein-corrected milk, on average). Diets did not affect N efficiency and nutrients digestibility. In the rumen, LIN+NIT did not affect protozoa number but reduced total volatile fatty acid (−12%) and propionate (−31%) concentrations. We concluded that linseed plus nitrate may have a long-term CH4-mitigating effect in dairy cows and that consuming milk products from cows fed nitrate may be safe in terms of nitrate and nitrite residuals. Further work is required to optimise the doses of linseed plus nitrate to avoid reduced cows performance.
Physiology and functional biology of systems
Research Article
The autophagy-related protein LC3 is processed in stallion spermatozoa during short-and long-term storage and the related stressful conditions
- I. M. Aparicio, P. Martin Muñoz, G. M. Salido, F. J. Peña, J. A. Tapia
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- 02 March 2016, pp. 1182-1191
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Use of cooled and frozen semen is becoming increasingly prevalent in the equine industry. However, these procedures cause harmful effects in the sperm cell resulting in reduced cell lifespan and fertility rates. Apoptosis and necrosis-related events are increased during semen cryopreservation. However, a third type of cell death, named autophagy, has not been studied during equine semen storage. Light chain (LC)3 protein is a key component of the autophagy pathway. Under autophagy activation, LC3-I is lipidated and converted to LC3-II. The ratio of LC3-II/LC3-I is widely used as a marker of autophagy activation. The main objective of this study was to investigate whether LC3 is processed during cooling, freezing and the stressful conditions associated with these technologies. A secondary objective was to determine if LC3 processing can be modulated and if that may improve the quality of cryopreserved semen. LC3 processing was studied by Western blot with a specific antibody that recognized both LC3-I and LC3-II. Viability was assessed by flow cytometry. Modulation of LC3-I to LC3-II was studied with known autophagy activators (STF-62247 and rapamycin) or inhibitors (chloroquine and 3-MA) used in somatic cells. The results showed that conversion of LC3-I to LC3-II increased significantly during cooling at 4°C, freezing/thawing and each of the stressful conditions tested (UV radiation, oxidative stress, osmotic stress and changes in temperature). STF-62247 and rapamycin increased the LC3-II/LC3-I ratio and decreased the viability of equine sperm, whereas chloroquine and 3-MA inhibited LC3 processing and maintained the percentage of viable cells after 2 h of incubation at 37°C. Finally, refrigeration at 4°C for 96 h and freezing at −196°C in the presence of chloroquine and 3-MA resulted in higher percentages of viable cells. In conclusion, results showed that an ‘autophagy-like’ mechanism may be involved in the regulation of sperm viability during equine semen cryopreservation. Modulation of autophagy during these reproductive technologies may result in an improvement of semen quality and therefore in higher fertility rates.
Relationships between ovulation rate and embryonic and placental characteristics in multiparous sows at 35 days of pregnancy
- C. L. A. Da Silva, H. van den Brand, B. F. A. Laurenssen, M. L. W. J Broekhuijse, E. F. Knol, B. Kemp, N. M. Soede
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- Published online by Cambridge University Press:
- 19 February 2016, pp. 1192-1199
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The objective of this study was to investigate relationships between ovulation rate (OR) and embryonic and placental development in sows. Topigs Norsvin® sows (n=91, parity 2 to 17) from three different genetic backgrounds were slaughtered at 35 days of pregnancy and the reproductive tract was collected. The corpora lutea (CL) were counted and the number of vital and non-vital embryos, embryonic spacing (distance between two embryos), implantation length, placental length, placental weight and embryonic weight were assessed. The difference between number of CL and total number of embryos was considered as early embryonic mortality. The number of non-vital embryos was considered as late mortality. Relationships between OR and all other variables were investigated using two models: the first considered parity as class effect (n=91) and the second used a subset of sows with parities 4 to 10 (n=47) to analyse the genetic background as class effect. OR was significantly affected by parity (P<0.0001), but was not affected by the genetic background of the sows. Parity and genetic background did not affect embryonic and placental characteristics at 35 days of pregnancy. OR (varying from 17 to 38 CL) was positively related with early embryonic mortality (β=0.49±0.1 n/ovulations, P<0.0001), with late embryonic mortality or number of non-vital embryos (β=0.24±0.1 n/ovulations, P=0.001) and with the number of vital embryos (β=0.26±0.1 n/ovulations, P=0.01). However, dividing OR in four classes, showed that the number of vital embryos was lowest in OR class 1 (17 to 21 CL), but not different for the other OR classes, suggesting a plateau for number of vital embryos for OR above 22. There was a negative linear relationship between OR and vital embryonic spacing (β=−0.45±0.1 cm/ovulation, P=0.001), implantation length (β=−0.35±0.1 cm/ovulation, P=0.003), placental length (β=−0.38±0.2 cm/ovulation, P=0.05) and empty space around embryonic-placental unit (β=−0.4±0.2 cm/ovulation, P=0.02), indicating uterine crowding. Further analyses showed that effects of OR on embryonic and uterine parameters were related with the increase in late mortality and not early embryonic mortality. Therefore, we conclude that a high OR results in an moderate increase in the number of vital embryos at day 35 of pregnancy, but compromises development in the surviving embryonic/placental units, suggesting that the future growth and survival of the embryos might be further compromised.
Short Communication
Restricted maternal nutrition alters myogenic regulatory factor expression in satellite cells of ovine offspring
- J. S. Raja, M. L. Hoffman, K. E. Govoni, S. A. Zinn, S. A. Reed
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- Published online by Cambridge University Press:
- 09 February 2016, pp. 1200-1203
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Poor maternal nutrition inhibits muscle development and postnatal muscle growth. Satellite cells are myogenic precursor cells that contribute to postnatal muscle growth, and their activity can be evaluated by the expression of several transcription factors. Paired-box (Pax)7 is expressed in quiescent and active satellite cells. MyoD is expressed in activated and proliferating satellite cells and myogenin is expressed in terminally differentiating cells. Disruption in the expression pattern or timing of expression of myogenic regulatory factors negatively affects muscle development and growth. We hypothesized that poor maternal nutrition during gestation would alter the in vitro temporal expression of MyoD and myogenin in satellite cells from offspring at birth and 3 months of age. Ewes were fed 100% or 60% of NRC requirements from day 31±1.3 of gestation. Lambs from control-fed (CON) or restricted-fed (RES) ewes were euthanized within 24 h of birth (birth; n=5) or were fed a control diet until 3 months of age (n=5). Satellite cells isolated from the semitendinosus muscle were used for gene expression analysis or cultured for 24, 48 or 72 h and immunostained for Pax7, MyoD or myogenin. Fusion index was calculated from a subset of cells allowed to differentiate. Compared with CON, temporal expression of MyoD and myogenin was altered in cultured satellite cells isolated from RES lambs at birth. The percent of cells expressing MyoD was greater in RES than CON (P=0.03) after 24 h in culture. After 48 h of culture, there was a greater percent of cells expressing myogenin in RES compared with CON (P<0.001). After 72 h of culture the percent of satellite cells expressing myogenin in RES was less than CON (P<0.01). There were no differences in the gene expression of Pax7, Myf5 or MyoD in isolated satellite cells at birth (P>0.05). In satellite cells from RES lambs at 3 months of age, the percent of cells expressing MyoD and myogenin were greater than CON after 72 h in culture (P<0.05). Fusion index was reduced in RES lambs at 3 months of age compared with CON (P<0.001). Restricted nutrition during gestation alters the temporal expression of myogenic regulatory factors in satellite cells of the offspring, which may reduce the pool of myoblasts, decrease myoblast fusion and contribute to the poor postnatal muscle growth previously observed in these animals.
Research Article
Lipopolysaccharide markedly changes glucose metabolism and mitochondrial function in the longissimus muscle of pigs
- H. Sun, Y. Huang, C. Yin, J. Guo, R. Zhao, X. Yang
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- 11 February 2016, pp. 1204-1212
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Most previous studies on the effects of lipopolysaccharide (LPS) in pigs focused on the body’s immune response, and few reports paid attention to body metabolism changes. To better understand the glucose metabolism changes in skeletal muscle following LPS challenge and to clarify the possible mechanism, 12 growing pigs were employed. Animals were treated with either 2 ml of saline or 15 µg/kg BW LPS, and samples were collected 6 h later. The glycolysis status and mitochondrial function in the longissimus dorsi (LD) muscle of pigs were analyzed. The results showed that serum lactate content and NADH content in LD muscle significantly increased compared with the control group. Most glycolysis-related genes expression, as well as hexokinase, pyruvate kinase and lactic dehydrogenase activity, in LD muscle was significantly higher compared with the control group. Mitochondrial complexes I and IV significantly increased, while mitochondrial ATP concentration markedly decreased. Significantly increased calcium content in the mitochondria was observed, and endoplasm reticulum (ER) stress has been demonstrated in the present study. The results showed that LPS treatment markedly changes glucose metabolism and mitochondrial function in the LD muscle of pigs, and increased calcium content induced by ER stress was possibly involved. The results provide new clues for clarifying metabolic diseases in muscle induced by LPS.
Behaviour, welfare and health
Short Communication
Circulating extracellular miR-22, miR-155, and miR-365 as candidate biomarkers to assess transport-related stress in turkeys
- C. Lecchi, A. T. Marques, M. Redegalli, S. Meani, L. J. Vinco, V. Bronzo, F. Ceciliani
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- 13 January 2016, pp. 1213-1217
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MicroRNA (miRNA) have been identified in circulating blood and might have the potential to be used as biomarkers for several pathophysiological conditions. To identify miRNA that are altered following stress events, turkeys (Meleagris gallopavo) were subjected to 2 h of road transportation. The expression levels of five circulating miRNA, namely miR-22, miR-155-5p, miR-181a-3p, miR-204 and miR-365-3p, were detected and assessed by quantitative polymerase chain reaction using TaqMan® probes, as potential biomarkers of stress. The areas under the receiver operating characteristic curves were then used to evaluate the diagnostic performance of miRNA. A panel of three stress-responsive miRNA, miR-22, miR-155 and miR-365 were identified; their expression levels were significantly higher after road transportation and the area under the curve (AUC) were 0.763, 0.71 and 0.704, respectively. Combining the three miRNA a specificity similar to the one found for the three miRNA separately was found. The AUC of the weighted average of the three miRNA was 0.763. This preliminary study suggests that the expression levels of circulating miR-22, miR-155 and miR-365 are increased during transport-related stress and that they may have diagnostic value to discriminate between stressed- and unstressed animals.
Research Article
Feather eating and its associations with plumage damage and feathers on the floor in commercial farms of laying hens
- A. B. Riber, L. K. Hinrichsen
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- Published online by Cambridge University Press:
- 02 February 2016, pp. 1218-1224
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Feather eating has been associated with feather pecking, which continues to pose economic and welfare problems in egg production. Knowledge on feather eating is limited and studies of feather eating in commercial flocks of laying hens have not been performed previously. Therefore, the main objective was to investigate feather eating and its association with plumage damage and floor feather characteristics in commercial flocks of layers in barn and organic production systems. The study was performed in 13 flocks of barn layers and 17 flocks of organic layers. Each flock was visited at around 32 and 62 weeks of age. During both visits, the plumage condition was assessed and the density of floor feathers recorded. In week 62, droppings and floor feathers were collected. Droppings were examined for presence of feather content, whereas length, downiness and pecking damage were recorded for each floor feather. In week 62, a higher prevalence of hens with poor plumage condition was found in barn (22.2%) compared with organic production systems (7.4%; P<0.001), but the prevalence of droppings with feather content did not differ between the two production systems (8.5% in barn v. 4.3% in organic; P=0.99). Our hypothesis about a positive correlation between feather eating and plumage damage was not supported as no correlation was found between the prevalence of poor plumage condition and the prevalence of droppings with feather content. However, the prevalence of pecking damaged floor feathers was positively correlated both with prevalence of droppings with feather content (P<0.05) and poor plumage condition (P<0.01), indicating a possible association between feather eating and feather pecking. In conclusion, it was confirmed that feather eating occurs on-farm, but feather eating was only found to be positively correlated to the number of floor feathers with pecking damage and not as expected to the prevalence of plumage damage. More research is needed into the sources from where feathers are selected for ingestion, that is, whether they are picked from the floor litter, plucked directly from other hens or dislodged during preening of own feathers.
Influence of social mixing and group size on skin lesions and mounting in organic entire male pigs
- R. Thomsen, S. A. Edwards, T. Rousing, R. Labouriau, J. T. Sørensen
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- Published online by Cambridge University Press:
- 09 February 2016, pp. 1225-1233
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Alternatives to surgical castration are needed, due to stress and pain caused by castration of male pigs. One alternative is production of entire male pigs. However, changed behaviour of entire males compared with castrated males might adversely affect the welfare of entire males and changes in management procedures and production system might be needed. Elements from the organic pig production system might be beneficial in this aspect. The aim of this article is to investigate the effect of grouping strategy including social mixing and group size on levels of mounting behaviour and skin lesions, hypothesising that procedures that disrupt the social stability (e.g. regrouping) will have a larger negative effect in small groups compared with large groups. Approximately 1600 organic entire male pigs of the breed (Landrace×Yorkshire)×Duroc were reared in parallel in five organic herds, distributed across four batches in a 2×2 factorial design in order to test the influence of social mixing (presence or absence of social mixing at relocation) and group size (15 and 30 animals). Animals were able to socialise with piglets from other litters during the lactation period, and were all mixed across litters at weaning. A second mixing occurred at insertion to fattening pens for pigs being regrouped. Counting of skin lesions (1348 or 1124 pigs) and registration of mounting behaviour (1434 or 1258 pigs) were done on two occasions during the experimental period. No interactive effects were found between social mixing and group size on either skin lesions or mounting frequency. Herd differences were found for both mounting frequency and number of skin lesions. No association between skin lesions and mounting were revealed. Social mixing and group size were shown as interacting effects with herds on mounting frequency (P<0.0001), but with no consistent pattern across all herds. In addition, no effect of social mixing was found on mean number of skin lesions, but more lesions were observed in large groups (P<0.036). This could indicate that keeping entire male pigs in groups of 30 animals as compared with smaller groups of 15 may marginally decrease the welfare of these animals.
Genetic associations between maternal traits and aggressive behaviour in Large White sows
- A. K. Appel, B. Voß, B. Tönepöhl, U. König von Borstel, M. Gauly
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- 12 February 2016, pp. 1234-1242
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The present study examined the possibilities and consequences of selecting pigs for reduced aggression and desirable maternal behaviour. Data were recorded from 798 purebred Large White gilts, with an age of 217±17.7 (mean±SD) days, which were observed at mixing with unfamiliar conspecifics. The reaction of the sows towards separation from their litter was assessed for 2022 litters from 848 Large White sows. Sows’ performance during their time in the farrowing unit was scored based on the traits farrowing behaviour (i.e. need of birth assistance), rearing performance (i.e. litter quality at day 10 postpartum (pp)), usability (i.e. additional labour input during lactation period e.g. for treatments) and udder quality of the sow (i.e. udder attachment). For agonistic behaviour, traits heritabilities of h2=0.11±0.04 to h2=0.28±0.06 were estimated. For the sow’s reaction towards separation from her litter low heritabilities were found (h2=0.03±0.03 for separation test on day 1 pp and h2=0.02±0.03 for separation test on day 10 pp). Heritabilities for lactating sow’s performance (farrowing behaviour, rearing performance, usability of the sow and udder quality) in the farrowing unit ranged from h2=0.03±0.02 to h2=0.19±0.03. Due to these results it can be assumed that selection for these traits, for example, for udder quality or reduced aggression, is possible. Antagonistic associations were found between separation test on day 1 pp and different measures of aggressiveness (rg=−0.22±0.26 aggressive attack and rg=−0.41±0.33 reciprocal fighting). Future studies should determine economic as well as welfare-related values of these traits in order to decide whether selection for these traits will be reasonable.