Heat Stress Reduces Intestinal Barrier Integrity and Favors Intestinal Glucose Transport in Growing Pigs

Excessive heat exposure reduces intestinal integrity and post-absorptive energetics that can inhibit wellbeing and be fatal. Therefore, our objectives were to examine how acute heat stress (HS) alters intestinal integrity and metabolism in growing pigs. Animals were exposed to either thermal neutral (TN, 21°C; 35–50% humidity; n = 8) or HS conditions (35°C; 24–43% humidity; n = 8) for 24 h. Compared to TN, rectal temperatures in HS pigs increased by 1.6°C and respiration rates by 2-fold (P<0.05). As expected, HS decreased feed intake by 53% (P<0.05) and body weight (P<0.05) compared to TN pigs. Ileum heat shock protein 70 expression increased (P<0.05), while intestinal integrity was compromised in the HS pigs (ileum and colon TER decreased; P<0.05). Furthermore, HS increased serum endotoxin concentrations (P = 0.05). Intestinal permeability was accompanied by an increase in protein expression of myosin light chain kinase (P<0.05) and casein kinase II-α (P = 0.06). Protein expression of tight junction (TJ) proteins in the ileum revealed claudin 3 and occludin expression to be increased overall due to HS (P<0.05), while there were no differences in claudin 1 expression. Intestinal glucose transport and blood glucose were elevated due to HS (P<0.05). This was supported by increased ileum Na⁺/K⁺ ATPase activity in HS pigs. SGLT-1 protein expression was unaltered; however, HS increased ileal GLUT-2 protein expression (P = 0.06). Altogether, these data indicate that HS reduce intestinal integrity and increase intestinal stress and glucose transport.     

Time course determination of the effects of rapid and gradual cooling after acute hyperthermia on body temperature and intestinal integrity in pigs

Rapid cooling after acute hyperthermia may cause a sustained increase in body temperature and exacerbate intestinal damage in pigs. Therefore, the study objective was to evaluate the temporal effects of rapid and gradual cooling on body temperature response and intestinal integrity after acute hyperthermia in pigs. In three repetitions, 54 pigs [83.3 ± 6.7 kg initial body weight (BW)], balanced by sex were exposed to thermoneutral conditions for 6 h (TN; n = 6 pigs/repetition; 21.1 ± 2.0°C), or heat stress conditions (HS; 39.3 ± 1.6°C) for 3 h, followed by a 3 h recovery period of gradual cooling [HSGC; n = 6 pigs/repetition; gradual decrease from HS to TN conditions] or rapid cooling [HSRC; n = 6 pigs/repetition; rapid TN exposure and cold water (4.0°C) dousing every 30 min for 1.5 h]. Feed was withheld throughout the entire 6 h period, but water was provided ad libitum. Gastrointestinal (T_GI) and rectal (T_R) temperatures were recorded every 15 min during the HS and recovery periods. Six pigs per repetition (n = 2/treatment) were euthanized and jejunal and ileal samples were collected for histology immediately after (d 0), 2 d after, and 4 d after the recovery period. Data were analyzed using PROC MIXED in SAS 9.4. Overall, rapid cooling reduced T_R and T_GI (P < 0.01; 0.95°C and 0.74°C, respectively) compared to gradual cooling. Jejunal villus height was reduced overall (P = 0.02; 14.01%) in HSGC compared to HSRC and TN pigs. Jejunal villus height-to-crypt depth ratio was reduced overall (P = 0.05; 16.76%) in HSGC compared to TN pigs. Ileal villus height was reduced overall (P < 0.01; 16.95%) in HSGC compared to HSRC and TN pigs. No other intestinal morphology differences were detected. In summary, HSRC did not cause a sustained increase in body temperature and did not negatively impact biomarkers of intestinal integrity in pigs.     

Involvement of Zn Depletion in Cd-Induced Toxicity on Prenatal Bone Formation in Rat

This experiment was conducted to evaluate the effects of chromium methionine with/without zinc sulfate or zinc amino acid complex on the growth performance, carcass traits, meat quality, serum parameters, endocrine parameters, and antioxidant status of growing-finishing pigs. A total of 180 (32.0 ± 1.7 kg body weight, BW) crossbred pigs (Duroc × Landrace × Yorkshire) were used in a completely randomized design with three dietary treatments and 10 replicates per treatment (five pens of barrows and five pens of gilts with six pigs per replicate). Three treatments were corn-soybean meal-based diets supplemented with 100 mg Zn/kg from zinc sulfate (ZnSO₄), 100 mg Zn/kg from ZnSO₄ + 0.2 mg Cr/kg from chromium methionine complex (CrMet), or 50 mg Zn/kg from ZnSO₄ + 50 mg Zn/kg from zinc amino acid complex (ZnAA) + 0.2 mg Cr/kg from CrMet, respectively. The experiment lasted 105 days, of which was divided into three stages including phase 1 (30 to 50 kg BW), phase 2 (50 to 80 kg BW), and phase 3 (80 to 110 kg BW). Results showed that supplementation with CrMet and ZnAA improved (P < 0.05) the feed conversion of the pigs in phase 2, phase 3, and the overall experiment. Hot carcass weight, dressing percentage, and a longissimus dorsi muscle area were increased (P < 0.05) in pigs fed with diets supplemented with both CrMet and ZnAA compared with pigs fed with diets containing only ZnSO₄ (P < 0.05). There was also an increase (P < 0.01) pH_24 h in the longissimus dorsi muscle in pigs fed with diets supplemented with CrMet and ZnAA. The concentration of serum glucose in pigs fed with diets containing CrMet and ZnAA was decreased (P < 0.05) compared with that in pigs fed with the diet containing ZnSO₄. Supplementation with CrMet and ZnAA increased (P < 0.05) the circulating levels of insulin and decreased (P < 0.05) cortisol. There was an increase (P < 0.05) in total serum antioxidant capacity and Cu/Zn superoxide dismutase activity as well as a decrease (P < 0.05) in serum malondialdehyde concentrations in pigs fed with diets supplemented with CrMet and ZnAA compared with pigs fed with the diet supplemented only with ZnSO₄. In conclusion, supplementation of CrMet only or CrMet together with ZnAA improved feed conversion, carcass traits, and meat quality in the growing-finishing pigs.     

Effects of rapid temperature fluctuations prior to breeding on reproductive efficiency in replacement gilts

Rapidly cooling pigs after heat stress (HS) results in a pathophysiological condition, and because rapid temperature fluctuations may be associated with reduced reproductive success in sows, it lends itself to the hypothesis that these conditions may be linked. Objectives were to determine the effects of rapid cooling on thermal response and future reproductive success in pigs. Thirty-six replacement gilts (137.8±0.9 kg BW) were estrus synchronized and then 14.1±0.4 d after estrus confirmation, pigs were exposed to thermoneutral conditions (TN; n=12; 19.7±0.9°C) for 6 h, or HS (36.3±0.5°C) for 3 h, followed by 3 h of rapid cooling (HSRC; n=12; immediate TN exposure and water dousing) or gradual cooling (HSGC; n=12; gradual decrease to TN conditions) repeated over 2 d. Vaginal (T_V) and gastrointestinal tract temperatures (T_GI) were obtained every 15 min, and blood was collected on d 1 and d 2 during the HS and recovery periods at 180 and 60 min, respectively. Pigs were bred 8.3±0.8 d after thermal treatments over 2 d. Reproductive tracts were collected and total fetus number and viability were recorded 28.0±0.8 d after insemination. HS increased T_V and T_GI (P=0.01; 0.98 °C) in HSRC and HSGC compared to TN pigs. During recovery, T_V was reduced from 15 to 105 min (P=0.01; 0.33 °C) in HSRC compared to HSGC pigs, but no overall differences in T_GI were detected (P<0.05; 39.67 °C). Rapid cooling increased (P<0.05) TNFα compared to HSGC and TN pigs during recovery-d 1 (55.2%), HS-d 2 (35.1%), and recovery-d 2 (64.9%). Viable fetuses tended to be reduced (P=0.08; 10.5%) and moribund fetuses tended to be increased (P=0.09; 159.3%) in HSRC compared to HSGC and TN pigs. In summary, rapid cooling prior to breeding may contribute to reduced fetal viability and reproductive success in pigs.     

Effects of taurine supplementation on productive performance,nutrient digestibility and gene expression of nutrient transporters in quails reared under heat stress

This study was conducted to examine the effects of dietary taurine supplementation on productive performance, nutrient digestibility, antioxidant status, and the gene expression of ileal nutrient transporters in laying quails reared under heat stress (HS). One hundred and eighty laying Japanese quails (Coturnix coturnix japonica) were fed a basal diet or basal diet supplemented with either 2.5 or 5 g of taurine per kg of diet, and reared at either 22 ± 2 °C for 24 h/d (thermoneutral, TN) or 34 ± 2 °C for 8 h/d (HS) for 12 weeks. The quails reared under HS consumed less feed, produced less egg, and had lower dry matter, organic matter and crude protein apparent digestibilities compared with the quails reared under the TN condition (P = 0.001). However, increasing taurine concentrations in the diet improved feed intake and egg production (P = 0.001), but also the apparent digestibilities (P ≤ 0.027) in quails reared under HS. The greater doses (5 g/kg) of taurine resulted in more responses. The quails reared under HS had greater serum and liver MDA concentrations (P = 0.0001) which decreased with dietary taurine supplementations, particularly greater doses. The gene expressions of ileal PEPT1, EAAT3, CAT1, CAT2, SGLT1, SGLT5, GLUT2, and GLUT5 decreased under HS conditions (P = 0.001). However, supplementing taurine, in a dose-dependent fashion, to the diet of quails reared under HS resulted in increases in the gene expressions of the transporters (P < 0.05) except for CAT1. The results of the present work showed that taurine supplementation, particularly with greater doses (5 g/kg), to the diet of laying quails kept under HS acts as alleviating negative effects of HS, resulting in improvements in productive performance and nutrient digestion, and also upregulation of ileal nutrient transporters.     

Heat stress of gilts around farrowing causes oxygen insufficiency in the umbilical cord and reduces piglet survival

Late gestating sows are susceptible to high ambient temperatures, possibly causing farrowing complications and reducing piglet survival. This experiment aimed to quantify in the days leading up to farrowing the impact of sow heat stress (HS) on farrowing physiology and survival of the piglets. Pregnant primiparous sows (gilts) were allocated to either thermoneutral control (CON, n = 8; constant 20 °C) or cyclical HS conditions (n = 8; 0900 h to 1700 h, 30 °C; 1700 h to 0900 h, 28 °C) from d 110 of gestation until farrowing completion. Gilt respiration rate, skin temperature and rectal temperature were recorded daily, and farrowing duration was quantified by video analyses. Blood samples were collected from the piglet umbilical vein at birth. At 48 h of age, piglet growth was quantified by morphometric analyses. The thermal exposure model induced HS and respiratory alkalosis in the gilts, as indicated by increased respiration rate, rectal temperature, skin temperature (all P < 0.001), plasma cortisol (P = 0.01) and blood pH (P < 0.001). Heat-stressed gilts took longer to start expelling placentae (P = 0.003), although the active farrowing duration was not significantly different between treatments. Stillbirth rates were higher in the HS group (P < 0.001), with surviving piglets at birth having lower umbilical vein partial pressure of oxygen (P = 0.04), oxygen saturation rate (P = 0.03) and tending to have increased lactate concentrations (P = 0.07). At birth, piglet skin meconium staining scores were greater in the HS group (P = 0.022). At 48 h of age, piglets from the HS group had reduced small intestinal length (P = 0.02), reduced jejunal crypt depth (P = 0.02) and lighter absolute brain weight (P = 0.001). In contrast, piglet BW, growth rate, relative organ weight and small intestinal mucosal barrier function did not change between treatments. Collectively, these findings demonstrated gilt HS during late gestation caused farrowing complications and reduced the umbilical oxygen supply to the piglets at parturition, leading to increased risks of piglet stillbirth with implications on impaired neonatal survivability and development.     

Effects of zinc source and dietary concentration on serum zinc concentrations,growth performance,wool and reproductive characteristics in developing rams

Dietary Zn has significant impacts on the growth and development of breeding rams. The objectives of this study were to evaluate the effects of dietary Zn source and concentration on serum Zn concentration, growth performance, wool traits and reproductive performance in rams. Forty-four Targhee rams (14 months; 68 ± 18 kg BW) were used in an 84-day completely randomized design and were fed one of three pelleted dietary treatments: (1) a control without fortified Zn (CON; n = 15; ~1 × NRC); (2) a diet fortified with a Zn amino acid complex (ZnAA; n = 14; ~2 × NRC) and (3) a diet fortified with ZnSO₄ (ZnSO₄; n = 15; ~2 × NRC). Growth and wool characteristics measured throughout the course of the study were BW, average daily gain (ADG), dry matter intake (DMI), feed efficiency (G : F), longissimus dorsi muscle depth (LMD), back fat (BF), wool staple length (SL) and average fibre diameter (AFD). Blood was collected from each ram at four time periods to quantify serum Zn and testosterone concentrations. Semen was collected 1 to 2 days after the trial was completed. There were no differences in BW (P = 0.45), DMI (P = 0.18), LMD (P = 0.48), BF (P = 0.47) and AFD (P = 0.9) among treatment groups. ZnSO₄ had greater (P ≤ 0.03) serum Zn concentrations compared with ZnAA and CON treatments. Rams consuming ZnAA had greater (P ≤ 0.03) ADG than ZnSO₄ and CON. There tended to be differences among groups for G : F (P = 0.06), with ZnAA being numerically greater than ZnSO₄ and CON. Wool staple length regrowth was greater (P < 0.001) in ZnSO₄ and tended to be longer (P = 0.06) in ZnAA treatment group compared with CON. No differences were observed among treatments in scrotal circumference, testosterone, spermatozoa concentration within ram semen, % motility, % live sperm and % sperm abnormalities (P ≥ 0.23). Results indicated beneficial effects of feeding increased Zn concentrations to developing Targhee rams, although Zn source elicited differential responses in performance characteristics measured.     

Preventive effects of supplemental dietary zinc on heat-induced damage in the epididymis of boars

Hyperthermia in boars reduces growth performance and sperm production. Zinc is an essential trace element in animal nutrition. Here we investigate the effects of dietary zinc on epididymal structure and function in Bama miniature pigs treated with heat exposure and investigate approaches to improve the reproductive performance in summer. Male Bama miniature pigs (n=18; aged 6 months; bodyweight=10.79±0.06 kg) were randomly allocated to 3 groups: control group (Control), heat treatment group (HT), and the diet-supplemented and heat treatment group (H+Zn). The Control and HT groups were fed with basal diet and the H+Zn group were fed with basal diet plus 1500 mg/kg zinc daily. After being fed with these 2 different diets for 30 days, pigs in the HT and H+Zn groups were exposed to 5 h of 40 °C heat treatment for 8 days. Rectal temperature and jugular venous blood were collected 3 h after onset of heat exposure on days 1, 4 and 8. Pigs were sacrificed after the termination of heat exposure. Heat treatment increased serum testosterone concentration on day 1 and 4 (P<0.01). In addition, the HT group displayed an increase in the clear cell count and a decrease in epithelium thickness in the caput epithelium (P<0.01, P<0.05), and dietary zinc protected the boars from these impairments (P<0.01, P=0.29). Evaluation of oxidative states showed that heat exposure increased the levels of malondialdehyde (MDA) and glutathione (GSH) in the epididymis (P<0.01, P<0.05), while dietary zinc reduced this elevation (P<0.01, P<0.01). Heat exposure enhanced the glucocorticoid receptor (GR) expression in the nuclei of principal and basal cells (P<0.01, P<0.01) while dietary zinc attenuated the GR immunoreactivity intensity (P<0.01, P<0.01). These results demonstrate that dietary zinc protects the epididymis from high temperature-induced impairment, alleviates oxidative stress, restores the integrity of the caput epithelium and decreases the stress response.     

Nonlinear analysis of heart rate variability for evaluating the growing pig stress response to an acute heat episode

Heart rate variability (HRV) is a proxy measure of autonomic function and can be used as an indicator of swine stress. While traditional linear measures are used to distinguish between stressed and unstressed treatments, inclusion of nonlinear HRV measures that evaluate data structure and organization shows promise for improving HRV interpretation. The objective of this study was to evaluate the inclusion of nonlinear HRV measures in response to an acute heat episode. Twenty 12- to 14-week-old growing pigs were individually housed for 7 days and acclimated to thermoneutral conditions (20.35°C ± 0.01°C; 67.6% ± 0.2% RH) before undergoing one of the two treatments: (1) thermoneutral control (TN; n = 10 pigs) or (2) acute heat stress (HS; n = 10 pigs; 32.6°C ± 0.1°C; 26.2% ± 0.1% RH). In Phase 1 of the experimental procedure (P1; 60 min), pigs underwent a baseline HRV measurement period in thermoneutral conditions before treatment [Phase 2; P2; 60 min once gastrointestinal temperature (T_g) reached 40.6°C], where HS pigs were exposed to heated conditions and TN pigs remained in thermoneutral conditions. After P2, all pigs were moved back to thermoneutral conditions (Phase 3; P3; 60 min). During each phase, T_g data were collected every 5 min and behavioural data were collected to evaluate the amount of time each pig spent in an active posture. Additionally, linear (time and frequency domain) and nonlinear [sample entropy (SampEn), de-trended fluctuation analysis, percentage recurrence, percentage determinism (%DET), mean diagonal line length in a recurrence plot] HRV measures were quantified. Heat stressed pigs exhibited greater T_g (P = 0.002) and spent less time in an active posture compared to TN pigs during P2 (P = 0.0003). Additionally, low frequency to high frequency ratio was greater in HS pigs during P3 compared to TN pigs (P = 0.02). SampEn was reduced in HS pigs during P2 (P = 0.01) and P3 (P = 0.03) compared to TN pigs. Heat stressed pigs exhibited greater %DET during P3 (P = 0.03) and tended to have greater %DET (P = 0.09) during P2 than TN pigs. No differences between treatments were detected for the remaining HRV measures. In conclusion, linear HRV measures were largely unchanged during P2. However, changes to SampEn and %DET suggest increased heat stress as a result of the acute heat episode. Future work should continue to evaluate the benefits of including nonlinear HRV measures in HRV analysis of swine heat stress.     

Pigs that are divergent in feed efficiency,differ in intestinal enzyme and nutrient transporter gene expression,nutrient digestibility and microbial activity

Feed efficiency is an important trait in the future sustainability of pig production, however, the mechanisms involved are not fully elucidated. The objective of this study was to examine nutrient digestibility, organ weights, select bacterial populations, volatile fatty acids (VFA’s), enzyme and intestinal nutrient transporter gene expression in a pig population divergent in feed efficiency. Male pigs (n=75; initial BW 22.4 kg SEM 2.03 kg) were fed a standard finishing diet for 43 days before slaughter to evaluate feed intake and growth for the purpose of calculating residual feed intake (RFI). Phenotypic RFI was calculated as the residuals from a regression model regressing average daily feed intake (ADFI) on average daily gain (ADG) and midtest BW^0.60 (MBW). On day 115, 16 pigs (85 kg SEM 2.8 kg), designated as high RFI (HRFI) and low RFI (LRFI) were slaughtered and digesta was collected to calculate the coefficient of apparent ileal digestibility (CAID), total tract nutrient digestibility (CATTD), microbial populations and VFA’s. Intestinal tissue was collected to examine intestinal nutrient transporter and enzyme gene expression. The LRFI pigs had lower ADFI (P<0.001), improved feed conversion ratio (P<0.001) and an improved RFI value relative to HRFI pigs (0.19 v. −0.14 SEM 0.08; P<0.001). The LRFI pigs had an increased CAID of gross energy (GE), and an improved CATTD of GE, nitrogen and dry matter compared to HRFI pigs (P<0.05). The LRFI pigs had higher relative gene expression levels of fatty acid binding transporter 2 (FABP2) (P<0.01), the sodium/glucose co-transporter 1 (SGLT1) (P<0.05), the glucose transporter GLUT2 (P<0.10), and the enzyme sucrase–isomaltase (SI) (P<0.05) in the jejunum. The LRFI pigs had increased populations of lactobacillus spp. in the caecum compared with HRFI pigs. In colonic digesta HRFI pigs had increased acetic acid concentrations (P<0.05). Differences in nutrient digestibility, intestinal microbial populations and gene expression levels of intestinal nutrient transporters could contribute to the biological processes responsible for feed efficiency in pigs.     

Effect of chronic cyclic heat stress on the intestinal morphology,oxidative status and cecal bacterial communities in broilers

The objective of this study was to examine the effects of chronic cyclic heat stress (HS) on the intestinal morphology, oxidative stress and cecal bacterial communities of broilers. One-day-old Arbor Acres (AA) male broilers (n = 100) were acclimated for 3 weeks and then randomly allocated into two groups, normal control (NC) group (22 ± 1 °C, 24 h/day) and HS group (32 ± 1 °C, 10 h/day lasted for 2 weeks). At 35 d of age, intestinal segments (duodenum, jejunum and ileum) and cecal digesta were collected for detection. HS affected intestinal morphology, inducing epithelial cell abscission, inflammatory cell infiltration, and lamina propria edema. Compared with the NC group, HS significantly decreased (P < 0.01) villus height (VH) and the VH-to-crypt depth (CD) ratio (VCR), increased (P < 0.05) CD in the duodenum and ileum, but had no effect on the VH in the jejunum. Moreover, HS induced oxidative stress with antioxidant enzymes activity decreasing (P < 0.05) while malondialdehyde (MDA) content increasing (P < 0.05) in small intestine. Pearson's correlation analysis indicated that MDA content was negatively correlated with VH (P < 0.05). The result of 16S rRNA sequencing showed that HS exposure impacted cecal microbiota alpha diversity (phylogenetic diversity whole-tree index) and beta diversity. Based on principal coordinate analysis (PCoA) plots for weighted UniFrac metrics and unweighted pair group method with arithmetic mean (UPGMA), there were 8 discriminative features at the genus level (linear discriminant analysis score > 2). Parabacteroides, Saccharimonas, Romboutsia and Weissella were reduced, while Anaerofustis, Pseudonocardia, Rikenella and Tyzzerella were enriched in heat-stressed broilers. Collectively, these results indicated that chronic cyclic HS induced oxidative stress that caused damage to intestinal villus-crypt structures, and then altered the cecal microflora profile.     

江山乌猪肠道黏膜菌群组成及其功能的性别差异

肠道黏膜微生物在调控宿主生理功能方面发挥重要作用,其结构组成受到多种因素影响。性别被认为是塑造肠道微生物的因素之一。然而,性别对肠道黏膜菌群的差异影响还不清楚。【目的】以江山乌猪为研究对象,探究性别差异对其肠道黏膜微生物组成及功能的影响。【方法】选取性成熟的雌性和雄性江山乌猪各8头,利用16S rRNA基因高通量测序技术分析回肠和结肠黏膜菌群。【结果】在回肠黏膜中,雄性江山乌猪菌群的Chao1指数和Shannon指数显著高于雌性(P<0.05),在结肠黏膜中,不同性别江山乌猪菌群Chao1指数和Shannon指数无显著差异(P>0.05)。菌群差异分析显示,回肠黏膜中,沙雷氏菌属(Serratia)和埃希氏志贺菌属(Escherichia_Shigella)在雌性组中的相对丰度显著高于雄性组(P<0.05),雄性组中Oscillospiraceae UCG-005、拟普雷沃氏菌属(Alloprevotella)、布劳特氏菌属(Blautia)和Prevotellaceae_NK3B31_group相对丰度显著高于雌性组(P<0.05);结肠黏膜中,雌性组中unclassified_Muribaculaceae、Rikenellaceae_RC9_gut_group和Prevotellaceae UCG-003相对丰度显著高于雄性组(P<0.05),Oscillospiraceae UCG-005、Lachnospiraceae_NK4A136_group和unclassified_Lachnospiraceae在雄性组中相对丰度更高(P<0.05)。功能预测发现,雄性乌猪回肠黏膜菌群显著富集了氨基酸代谢、碳水化合物代谢和能量代谢等功能途径(P<0.05);结肠黏膜菌群主要富集了膜转运相关的ABC转运蛋白和信号转导相关的双组分系统等功能途径(P<0.05)。【结论】不同性别江山乌猪肠黏膜菌群结构及功能具有明显差异。这些结果揭示了不同性别江山乌猪肠道黏膜菌群的差异特征,为了解和挖掘我国地方畜禽品种肠道微生物资源提供部分参考。     

In vitro supplementation with the porcine plasma product,betaGRO®, stimulates activity of porcine fetal myoblasts and neonatal satellite cells in a divergent manner

Two separate experiments were conducted to evaluate the effect of betaGRO® supplementation on in vitro porcine fetal myoblasts (PFM) and porcine satellite cells (PSC) proliferation, fusion and myotube thickness. The PFM and PSC were isolated from the m. longissimus dorsi of day 60 of gestation fetuses and piglets within 24 h of birth, respectively. Proliferation assays were conducted as 4×3 factorial arrangements with time of culture (24, 48, 72, 96 h) and media treatment (standard porcine media supplemented with 10% (vol/vol) fetal bovine serum (HS); HS without 10% fetal bovine serum (LS); and LS supplemented with 10 mg/ml betaGRO® (BG)) as main effects. Fusion and myotube growth assays were conducted as 2×2 factorial designs with serum concentration (HS or LS), and betaGRO® inclusion (0 or 10 mg/ml) as main effects. There was a treatment×time interaction and betaGRO®×serum interactions for proliferation, fusion and myotube thickness of PFM (P<0.01). At all-time points, HS and BG-PFM had greater proliferation rates compared LS (P<0.01). The HS treatment had greater proliferation rates than BG (P<0.02) except at 72 h of culture (P=0.44). When betaGRO® was added to LS media, fusion percentage and myotube thickness decreased (P<0.01), while fusion percentage increased (P<0.01) and myotube thickness was unaffected (P=0.63) when betaGRO® was added to HS media. There were treatment×time and betaGRO®×serum interactions for proliferation rate and fusion rate of PSC, respectively (P<0.01). At all-time points, HS had greater proliferation rates than LS and BG (P<0.01), and LS had greater proliferation rates than BG (P<0.02). When betaGRO® was added to LS and HS media, fusion percentage increased for both media types (P<0.01). There was no betaGRO®×serum interaction (P=0.63) for PSC myotube thickness; however, betaGRO® supplemented myotubes were thicker (P<0.01) than non-betaGRO® supplemented myotubes. These two experiments indicate in vitro betaGRO® supplementation stimulates divergent responses based on the age of cell examined.     

Effects of floor type,stocking density,slaughter age and gender on productive and qualitative traits of rabbits reared in collective pens

At 34 days of age, 376 crossbred rabbits of both sexes were housed in 16 open-top collective pens (1.68 m²) according to a 2×2×2 factorial arrangement with two types of pen floor (plastic v. wooden slatted), two stocking densities (12 v. 16 animals/m²) and two slaughter ages (76 v. 83 days). The rabbits were examined for growth performance, slaughter results and meat quality. The effect of gender was also examined. The percentage of rabbits with wounds due to aggression varied with stocking density (8.2% v. 26.2% for 12 v. 16 animals/m²; P⩽0.001), slaughter age (15.0% v. 22.0% at 76 v. 83 days; P⩽0.10) and gender (11.3% v. 25.8% for females v. males; P⩽0.001). Rearing rabbits on a plastic rather than a wooden slatted floor promoted slaughter weight (2795 v. 2567 g; P⩽0.001), dressing percentage (61.4% v. 60.9%; P⩽0.01), dissectible fat (2.4% v. 2.0%; P⩽0.01) and hind leg muscle-to-bone ratio (5.81 v. 5.35; P⩽0.001). Increased stocking density impaired daily growth (38.5 v. 35.9 g/day; P⩽0.05) and feed intake (140 v. 134 g/day; P⩽0.01) during the second period (55 days to slaughter) and decreased slaughter weight (2725 v. 2637 g; P⩽0.01). At the older slaughter age, the feed conversion ratio was impaired (2.98 v. 3.18; P⩽0.001); the slaughter weight (2574 g v. 2788 g; P⩽0.001), dissectible fat (2.0% v. 2.4%; P⩽0.01) and hind leg muscle-to-bone ratio (5.41 v. 5.75; P⩽0.01) increased; meat thawing losses, cooking losses and shear force decreased (P⩽0.05). The main differences between the females and males were found in the slaughter for transport losses (2.6% v. 2.2%; P⩽0.01) and longissimus lumborum proportions (13.0% v. 12.4%; P⩽0.01). In conclusion, the growth performance of pen-housed rabbits was largely determined by the type of floor and less affected by stocking density. The meat quality depended on ontogenetic factors, such as slaughter age and gender, and not on housing conditions. The differences in the percentages of wounded animals owing to experimental factors deserve further investigation from the perspective of animal welfare issues.     

Effects of Added Zinc on Skeletal Muscle Morphometrics and Gene Expression of Finishing Pigs Fed Ractopamine-HCL

Finishing pigs (n = 320) were used in a 35-day study to determine the effects of ractopamine-HCl (RAC) and supplemental Zinc (Zn) level on loin eye area (LEA) and gene expression. Pens were randomly allotted to the following treatments for the final 35 days on feed: a corn-soybean meal diet (CON), a diet with 10 ppm RAC (RAC+), and RAC diet plus added Zn at 75, 150, or 225 ppm. Sixteen pigs per treatment were randomly selected for collection of serial muscle biopsies and carcass data on day 0, 8, 18, and 32 of the treatment phase. Compared to CON carcasses, RAC+ carcasses had 12.6% larger (P = 0.03) LEA. Carcasses from RAC diets with added Zn had a tendency for increased (quadratic, P < 0.10) LEA compared to the RAC+ carcasses. Compared to RAC+ pigs, relative expression of IGF1 decreased with increasing levels of Zn on day 8 and 18 of treatment, but expression levels were similar on day 32 due to Zn treatments increasing in expression while the RAC+ treatment decreased (Zn quadratic × day quadratic, P = 0.04). A similar trend was detected for the expression of β₁-receptor where expression levels in the RAC+ pigs were greater than Zn supplemented pigs on day 8 and 18 of the experiment, but the magnitude of difference between the treatments was reduced on day 32 due to a decrease in expression by RAC+ pigs and an increase in expression by the Zn pigs (Zn quadratic × day quadratic, P = 0.01). The ability of Zn to prolong the expression of these two genes may be responsible for the tendency of Zn to increase LEA in RAC supplemented pigs.     

Analysis of apoptosis and DNA damage in bovine cumulus cells after exposure in vitro to different zinc concentrations

The purpose of this study was to investigate the effect of Zn (zinc) concentration on CCs (cumulus cells) during in vitro maturation. For this purpose, DNA integrity of CCs by addition of different Zn concentrations [0 (control); 0.7 μg/ml (Zn1); 1.1 μg/ml (Zn2) and 1.5 μg/ml (Zn3)] to the culture medium was evaluated by comet assay. In addition, early apoptosis was analysed by annexin staining assay. CCs treated with Zn showed a significant decrease in the DNA damage in a dose‐dependent manner. Comet assay analysed for TM (tail moment) was significantly higher in cells cultured without Zn (control, P<0.01) with respect to cells treated with Zn (control: 5.24±16.05; Zn1: 1.13±5.31; Zn2: 0.10±0.36; Zn3: 0.017±0.06). All treatments were statistically different from the control (P=0.014 for Zn1; P<0.01 for Zn2 and Zn3). The frequency of apoptotic cells was higher in the control group (control: 0.142±0.07; Zn1: 0.109±0.0328; Zn2:0.102±0.013; Zn3: 0.0577±0.019). Statistical differences were found between control and Zn1 (P=0.0308), control and Zn2 (P=0.0077), control and Zn3 (P<0.0001), Zn1 and Zn3 (P<0.001) and Zn2 and Zn3 (P=0.0004). No differences were found between Zn1 and Zn2. In conclusion, low Zn concentrations increase DNA damage and apoptosis in CCs cultured in vitro. However, adequate Zn concentrations ‘protect’ the integrity of DNA molecule and diminish the percentage of apoptotic CC.     

Nutrient restriction and realimentation in beef cows during early and mid-gestation and maternal and fetal hepatic and small intestinal in vitro oxygen consumption

Objectives were to determine the effects of advancing gestation, maternal nutrient restriction during early and mid-gestation, and realimentation on fetal liver and jejunal mass and energy use in both dams and fetuses. On day 30 of pregnancy, multiparous, non-lactating beef cows (initial BW=621±11.3 kg and body condition score=5.1±0.1) were assigned to one of the two dietary treatments: control (CON; 100% requirements; n=18) and restricted (R; 60% requirements; n=28). On day 85, cows were slaughtered (CON, n=6; R, n=6), and remaining cows continued on control (CC; n=12) and restricted (RR; n=12) diets, or were realimented to the control diet (RC; n=11). On day 140, cows were slaughtered (CC, n=6; RR, n=6; RC, n=5), remaining cows continued on the control diet (CCC, n=6; RCC, n=5), or were realimented to the control diet (RRC, n=6). On day 254, all remaining cows were slaughtered. Maternal liver O₂ consumption linearly increased (P⩽0.04) and jejunal weight (g/kg) linearly decreased (P=0.04) as gestation advanced in CON groups. Fetal BW, and hepatic and small intestinal absolute mass, protein content and O₂ consumption linearly increased (P⩽0.04) as pregnancy advanced in CON groups. However, mass and O₂ consumption relative to BW linearly decreased (P⩽0.001) in the fetal liver in CON groups. When analyzing the effects of dietary treatment, at day 85, fetal jejunal O₂ consumption (mol/min per kg BW) was lower (P=0.02) in the R group when compared with the CON group. At day 140, maternal hepatic weight (g) was lower (P=0.02) in RC and RR cows when compared with CC, and fetal jejunual O₂ consumption (mmol/min per mg tissue and mmol/min per g protein) was greater (P⩽0.02) in RC when compared with RR. At day 254, maternal hepatic O₂ consumption (absolute and relative to BW) was lower (P⩽0.04) in the RCC cows when compared with RRC. Fetal hepatic weight was lower (P=0.05) in the CCC group when compared with RCC and RRC. The changes in response to nutrient restriction and realimentation in both the dam and fetus may indicate an adaptation to a lower amount of available nutrients by altering tissue mass and metabolism.     

Effects of whole linseed and rumen-protected conjugated linoleic acid enriched diets on beef quality

Instrumental assessments and sensory tests were performed to evaluate the effects of diet and postmortem ageing time (1, 7 and 21 days) on beef quality. A total of 48 Friesian calves were randomly allocated to four dietary treatments: control, whole linseed (10% linseed), conjugated linoleic acid (CLA) (2% protected CLA), and whole linseed+CLA (10% linseed and 2% protected CLA). Animals were slaughtered at 458±16.6 kg live weight and 11 months of age. Ageing was more significant than diet on most instrumental parameters. Meat from linseed enriched diets had greater drip loss (P⩽0.001) and intramuscular fat (P⩽0.01) than meat from animals fed CLA. Beef aged for 7 and 21 days had lower cooking losses (P⩽0.01) and shear force (P⩽0.001) than beef aged for 1 day. Lightness was affected only by display time. The addition of CLA in the diet increased hue and yellowness, whereas the inclusion of linseed decreased these values, as well as increased redness. Linseed in the diet decreased fat odour (P⩽0.05), but increased beef (P⩽0.01) and liver (P⩽0.05) flavours. Meat aged for 21 days was significantly more rancid (P⩽0.001), even under vacuum storage. Several organoleptic properties were improved with the inclusion of linseed in the diet, whereas they remained unaffected by the inclusion of CLA.     

Lipid metabolism in pigs fed supplemental conjugated linoleic acid and/or dietary arginine

We proposed that the combination of conjugated linoleic acid (CLA) and arginine would decrease adiposity by depressing lipid synthesis in liver and adipose tissues of growing pigs. Pigs were allotted to treatments in a 2 × 2 factorial design with two lipids (CLA or canola oil) and two amino acids [l-arginine or l-alanine (isonitrogenous control)]; supplements were provided from 80 to 110 kg body weight (approximately 4 weeks). Treatment groups (n = 4) were: control (2.05% l-alanine plus 1% canola oil); CLA (2.05% l-alanine plus 1% CLA); arginine (1.0% l-arginine plus 1.0% canola oil); arginine plus CLA (1.0% arginine plus 1.0% CLA). Arginine increased backfat thickness (P = 0.07) in the absence or presence of CLA, and arginine supplementation increased subcutaneous and retroperitoneal adipocyte volume, especially in combination with dietary CLA (interaction P = 0.001). Arginine increased palmitate incorporation into total lipids by over 60% in liver (P = 0.07). Dietary CLA increased palmitate incorporation into lipids in longissimus muscle by over 100% (P = 0.01), and CLA increased longissimus muscle lipid by nearly 20%. CLA increased glucose oxidation to CO₂ by over 80% in retroperitoneal and subcutaneous adipose tissues (P = 0.04), and doubled palmitate oxidation to CO₂ in intestinal duodenal mucosal cells (P = 0.07). Arginine supplementation decreased muscle pH at 45 min postmortem (P = 0.001), indicating elevated early postmortem glycolysis, and CLA and arginine independently increased PGC-1α gene expression in longissimus muscle. CLA but not arginine depressed mTOR gene expression in intestinal duodenal mucosal cells. CLA decreased serum insulin by 50% (P = 0.02) but increased serum triacylglycerols by over 40%. CLA supplementation increased (P ≤ 0.01) total saturated fatty acids in liver and adipose tissue. In conclusion, neither CLA nor arginine depressed tissue lipid synthesis in growing/finishing pigs, and in fact dietary CLA promoted elevated intramuscular lipid and arginine increased carcass adiposity.