Deoxynivalenol induces oxidative stress,inflammatory response and apoptosis in bovine mammary epithelial cells

Deoxynivalenol (DON) is a toxic secondary metabolite produced by Fusarium graminearum. It is one of the most common feed contaminants that poses a serious threat to the health and performance of dairy cows. This study investigated the in vitro cytotoxicity of DON on bovine mammary epithelial cells (MAC‐T). DON at different concentrations (0.25, 0.3, 0.5, 0.8, 1 or 2 μg/ml) inhibited the growth of MAC‐T cells after 24 hr of exposure (p < .001). DON at 0.25 μg/ml increased lactate dehydrogenase (LDH) leakage (p < .05); decreased glutathione (GSH) levels (p < .001), total superoxide dismutase (T‐SOD) activity and total antioxidant capacity (T‐AOC; p < .01); and increased malondialdehyde (MDA) concentration (p < .01) in MAC‐T cells after 24 hr of exposure. We also observed that DON increased reactive oxygen species (ROS) levels in cells incubated for 9, 15 and 24 hr (p < .001). DON at 0.25 μg/ml triggered oxidative damage in MAC‐T cells. Furthermore, it induced an inflammatory response in the cells incubated for 9, 15 and 24 hr (p < .05) by increasing the mRNA expression levels of nuclear factor kappa B, myeloid differentiation factor 88 (MyD88), tumour necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β), IL‐6, cyclooxygenase‐2 and IL‐8. We further examined the effect of DON on apoptosis. DON prevented normal proliferation of MAC‐T cells by blocked cell cycle progression in 24 hr (p < .001). In addition, the apoptosis rate measured using annexin V‐FITC significantly increased (p < .05) with increase in the mRNA expression level of Bax (p < .01) and increase in the Bax/Bcl‐2 ratio (p < .01) in cells incubated for 24 hr. In summary, DON exerts toxic effects in MAC‐T cells by causing oxidative stress, inducing an inflammatory response, affecting cell cycle and leading to apoptosis.     

Use of protocatechuic acid as the sole antioxidant in the base medium for in vitro culture of ovine isolated secondary follicles

This study evaluated the effect of the protocatechuic acid (PCA) as the sole antioxidant in the base medium for in vitro culture of ovine secondary follicles. Secondary follicles (200‐230 μm) were isolated and cultured in α‐minimal essential medium supplemented with BSA, insulin, glutamine and hypoxanthine (α‐MEM: antioxidant‐free medium) or α‐MEM also added by transferrin, selenium and ascorbic acid (α‐MEM+: with antioxidant) or α‐MEM added by PCA (56.25; 112.5; 225; 450; or 900 μg/ml). Moreover, after culture, oocytes were matured and the chromatin configuration and DNA fragmentation were evaluated. After 12 days, the treatment containing 56.25 μg/ml PCA showed higher percentage of normal follicles than control medium or the other treatments (p < .05), except for 900 μg/ml PCA (p > .05). The antrum formation was significantly higher in treatments containing 56.25, 112.5 or 900 μg/ml PCA, compared to the α‐MEM and similar (p > .05) to the other treatments. The rates of fully grown oocytes (≥110 μm) were similar (p > .05) among all treatments containing PCA and α‐MEM+, and those were superior (p < .05) than α‐MEM, except for 450 μg/ml PCA (p > .05). GSH levels and mitochondrial activity were higher (p < .05) in α‐MEM+ than in α‐MEM and similar (p > .05) to all PCA treatments. The rates of meiotic resumption and DNA fragmentation were similar (p > .05) among α‐MEM+ and 56.25 μg/ml PCA. In conclusion, PCA at 56.25 μg/ml as the sole antioxidant added to the medium for ovine isolated secondary follicle culture maintains follicular survival, GSH and active mitochondria levels, meiotic developmental competence and DNA integrity of cultured oocytes.     

Effects of E2 binding enzyme UBC9 on porcine oocyte maturation,apoptosis and embryo development

SUMOylation is a dynamic post-translational modification process. However, the function of small ubiquitin-like modifiers (SUMOs) in the maturation of porcine oocytes and embryo growth is not well known. Therefore, the aim of this study was to investigate the effect of E2 binding enzyme UBC9 on the expression of SUMO-1 protein during the in vitro maturation of porcine oocytes and embryo development after in vitro fertilization. Four groups were used: 0 (Control), 5, 10 and 15 µg/ml UBC9. Western blotting, flow cytometry and RT-qPCR were used to detect the in vitro maturation of porcine oocytes, SUMO-1 content, viability and the expression of apoptotic genes. Compared to those in the control treatment, the maturation rate (p < .05) and viability (p < .01) of oocytes in the 5 μg/ml treatment group decreased significantly. SUMO-1 protein markers appeared at 59 and 71 kDa and the content of SUMO-1 protein in the 10 µg/ml treatment group decreased significantly (p < .05). In the expression of apoptosis-related genes, Bcl-2 gene expression was significantly downregulated in the 10 μg/ml treatment group (p < .05). However, Bax and Caspase-3 were significantly upregulated in the 5 μg/ml treatment group (p < .05). During embryonic development, the cleavage rate of oocytes in the 10 µg/ml treatment group was significantly reduced (p < .05), whereas blastocyst formation rate in the 5 µg/ml treatment group was significantly reduced. UBC9 regulates SUMO-1 content in mature pig oocytes in vitro, which affects oocyte maturation rate, viability, apoptotic genes expression and embryo development after fertilization.     

Effects of polymyxin‐B on TNF‐α production in equine whole blood stimulated with three different bacterial toxins

Polymyxin‐B is used to treat equine systemic inflammation. Bacterial toxins other than lipopolysaccharide (LPS) contribute to systemic inflammation but the effects of polymyxin‐B on these are poorly defined. Whole blood aliquots from six healthy horses diluted 1:1 with RPMI were incubated for 21 hr with 1 μg/ml of LPS, lipoteichoic acid (LTA) or peptidoglycan (PGN) in the presence of increasing concentrations of polymyxin‐B (10–3000 μg/ml). A murine L929 fibroblast bioassay was used to measure TNF‐α activity. Polymyxin‐B significantly inhibited the effects of all three bacterial toxins. Analysis of variance showed the IC₅₀ value for polymyxin‐B for TNF‐α inhibition caused by LTA (11.19 ± 2.89 μg/ml polymyxin‐B) was significantly lower (p = .009) than the values for LPS (46.48 ± 9.93 μg/ml) and PGN (54.44 ± 8.97 μg/ml). There was no significant difference in IC₅₀ values between LPS and PGN (p > .05). Maximum inhibition of TNF‐α was 77.4%, 73.0% and 82.7% for LPS, PGN and LTA, respectively and was not significantly different between toxins. At the two highest concentrations of polymyxin‐B, TNF‐α began to increase. These data suggest that polymyxin‐B may inhibit the effects of bacterial toxins other than LPS and might be a more potent inhibitor of LTA than LPS or PGN.     

Effect of dietary supplementation of calcium butyrate on growth performance,carcass traits,intestinal health and pro‐inflammatory cytokines in Japanese quails

The objective of the present study was to evaluate the potential effect of dietary calcium butyrate on growth performance, carcass traits and gut health in Japanese quails. In total, 320 one‐day‐old Japanese quails were randomly assigned to 4 equal treatments, with 8 replicates of 10 Japanese quails, for 4 weeks. The Japanese quails in control treatment were fed control diet whereas in the other treatments the Japanese quails were fed diet supplemented with calcium butyrate at 0.3, 0.5 and 0.7 g/kg diet. Data concerning performance measurements were recorded weekly. In addition, eight Japanese quails (one/replicate) from each treatment were selected randomly for serum collection to measure pro‐ and anti‐inflammatory cytokines. Pooled faecal samples from each replicate of each treatment were also collected at three time points (0, 2 and 4 weeks) for count E. coli and C. perfringens. The results showed that after 7 days of the experimental period, Japanese quails fed calcium butyrate supplemented diet at 0.7 g/kg showed a greater (p < .05) body weight and a favourable (p < .05) feed conversion ratio than the other treatments. Moreover, serum superoxide dismutase and catalase activities were increased (p < .05) in Japanese quails fed calcium butyrate supplemented diet at 0.7 g/kg. Calcium butyrate supplementation at 0.7 g/kg was associated with reduction (p < .05) in TNF‐α, IL‐6 and IL1‐β, while IL‐10 was increased (p < .05). In addition, after 2 weeks of calcium butyrate supplementation, a reduction (p < .05) in E. coli and C. perfringens counts was observed in excreta of Japanese quails fed 0.5 and 0.7 g calcium butyrate/kg diets. It is concluded that calcium butyrate supplementation improves body weight gain, reduces E. coli and C. perfringens counts and has anti‐inflammatory/anti‐oxidant effect in Japanese quails.     

Lactobacillus rhamnosus GG promotes M1 polarization in murine bone marrow‐derived macrophages by activating TLR2/MyD88/MAPK signaling pathway

Lactobacillus rhamnosus GG (LGG) is increasingly applied in functional food products and acts as a probiotic model in nutritious and clinical studies. Increasing evidences have revealed the immune modulation of LGG on macrophages. The aim of this study is to investigate the effect of LGG on macrophage polarization of murine bone marrow‐derived macrophages (BMDMs). BMDMs were treated with 10⁸ colony‐forming units (CFU)/ml LGG for 1.5, 3, and 6 hr. Results showed that LGG obviously upregulated the mRNA expression of M1‐associated cytokines (p < .05), including interleukin‐1 beta (IL‐1β), IL‐6, tumor necrosis factor‐alpha (TNF‐α), and inducible nitric oxide synthase (iNOS), whereas had no effect on the expression of M2‐associated markers (p > .05), including arginase 1 (Arg1), mannose receptor, and chitinase‐like protein 3 (YM1). Furthermore, LGG markedly increased the expression of pro‐inflammatory cytokines (IL‐12p40, cyclooxygenase‐2 [COX‐2], and interferon‐γ [IFN‐γ]) (p < .05) and anti‐inflammatory cytokines (IL‐10, IL‐4, and transforming growth factor‐β [TGF‐β]) (p < .05). In addition, we also found that TLR2/MyD88/MAPK signaling pathway was required for LGG‐induced M1 macrophage polarization and M1‐related cytokines expression. Together, these findings demonstrate that probiotic LGG facilitates M1 polarization of BMDMs, suggesting that LGG may have an immunotherapeutic potential in regulating the host defense against pathogen invasion.     

Changes of leukocyte counts and expression of pro‐ and anti‐inflammatory cytokines in peripheral leukocytes in periparturient dairy cows with retained fetal membranes

In dairy cows, retained fetal membranes (RFM) affect reproductive performance. The aim of this study was to examine the leukocyte counts and the gene expression of tumour necrosis factor α (TNFα), interleukin 1β (IL‐1β), IL‐8, and IL‐10 in polymorphonuclear leukocytes (PMNs) and peripheral blood mononuclear cells (PBMCs) in cows with (n = 5) or without (n = 5) RFM during the peripartum period. The lymphocyte counts in RFM cows were higher than those in control cows throughout the experiment (p < .05). The expression of IL‐8 in PMNs of control cows was higher (p < .05) compared with that of RFM cows postpartum. In cows with RFM, IL‐1β expression was higher (p < .05) in PMNs at 6 weeks postpartum whereas the expression of IL‐1β was lower (p < .05) in PBMCs at 4 weeks postpartum. The expression of IL‐10 in PBMCs of control cows was higher (p < .05) than that of RFM cows at 2 weeks prepartum and 4 weeks postpartum. Taken together, our data indicate that changes of gene expression of pro‐ and anti‐inflammatory cytokines in RFM cows might be associated with the delayed placental separation and development of uterine inflammation in RFM cows.     

Beta‐glucan from Agrobacterium sp. ZX09 improves growth performance and intestinal function in weaned piglets

Beta‐glucan is currently under consideration as an alternative to in‐feed antibiotics. The aim of the study was to investigate Agrobacterium sp. ZX09 beta‐glucan on intestinal morphology, cytokine concentration, mucin expression and microbial populations of weaning piglets. Pigs were randomly assigned to one of five dietary treatments supplemented with 0, 25, 50, 100 and 200 mg/kg beta‐glucan. Data showed an increase in ADG at the 100 mg/kg group (p = .03). A significant increase in villus height and reduction in crypt depth were fund in ileal tissue at the 100 mg/kg inclusion level (p < .05). Dietary supplementation of 100 mg/kg beta‐glucan enhanced IL‐10 concentration (p = .04) and gene expression of MUC1 and MUC2 (p < .05) in the jejunum. Dietary supplementation of 100 mg/kg beta‐glucan provoked the up‐regulation of Lactobacillus counts and down‐regulation of Escherichia coli counts in the caecum (p = .05). Data suggested that improved growth performance in response to beta‐glucan supplementation at 100 mg/kg in weaned piglets may be explained by the improved intestinal function.     

Effect of probiotic supplementation on growth performance,intestinal morphology,barrier integrity,and inflammatory response in broilers subjected to cyclic heat stress

This study investigated the protective effects of probiotic on heat stress‐induced intestinal injury and inflammatory response in broilers. A total of 180 male broilers were randomly allocated to three treatments with four replicates each from 22 to 42 days of age. The broilers were either raised under thermoneutral (TN) conditions (23 ± 1°C) or subjected to cyclic heat stress (28–35–28°C for 12 hr daily). The broilers kept at TN conditions were fed a basal diet, and those exposed to heat stress were fed basal diets supplemented with or without probiotic at a dose of 1.5 × 10⁸ cfu/kg. Compared with the TN group, heat stress decreased (p < .05) the growth performance, reduced (p < .05) villus height and villus height: crypt depth ratio in intestinal mucosa, increased (p < .05) serum levels of D‐lactic acid on day 28 and endotoxin, TNF‐α and IL‐6 on day 42, and decreased (p < .05) serum IL‐10 content on day 42. Dietary supplementation of probiotic reversed (p < .05) all these changes except for the growth performance in heat‐stressed broilers. In conclusion, dietary inclusion of probiotic could improve intestinal morphology and barrier function, alleviate inflammatory response, but exert no ameliorative effect on growth performance of broilers under cyclic heat stress.     

Bovine colostrum enriched with lyophilized bovine colostrum stimulates intestinal epithelium renewal of Holstein calves in the first days of life

Consumption of a second meal of colostrum with high quality could contribute to the intestinal epithelium development, especially if there is poor supply of colostrum just after birth. The effect of a second colostrum meal was evaluated on histomorphometry of the intestinal mucosa of newborn Holstein calves fed with high‐ and low‐quality first colostrum. Seventy‐two calves were fed with a first colostrum meal with high (HFM, close to 100 mg/ml) or low (LFM, close to 30 mg/ml) IgG concentration. At 12 hr of life, three treatments of second colostrum feeding were applied to the calves either fed high or low first colostrum: calves fed with low (LOW—close to 30 mg/ml) or high (HIGH—close to 100 mg/ml) IgG concentration; and colostrum enriched with lyophilized bovine colostrum with high IgG concentration (ENRICHED—higher than 120 mg/ml), resulting in six groups. Intestinal samples were collected after 24 and 72 hr of life. In the distal jejunum and ileum, LOW showed higher villus height than ENRICHED (p < .05). In the distal jejunum, greater villus perimeter was observed in the LOW compared to ENRICHED at 24 hr (p < .05). In ileum, LFM showed higher villus perimeter compared to HFM (p < .05). LOW showed the highest villus height‐to‐crypt depth ratio in the medium and distal jejunum and ileum, p < .05. ENRICHED and HFM showed decreased muscle layer thickness in the proximal and distal jejunum respectively (p < .05). The results reveal that the high concentration of total solids, crude protein, IgG and IGF‐I of colostrum with high quality worsened the absorptive area, but may have stimulated the activity of cell division in intestinal crypts. Considering the present results, bovine colostrum enriched with lyophilized bovine colostrum stimulates intestinal epithelium renewal of Holstein calves in the first days of life.     

Crocin supplementation during oocyte maturation enhances antioxidant defence and subsequent cleavage rate

The purpose of the present study was to assess the effect of crocin supplementation during oocyte maturation on the antioxidant defence and anti‐apoptotic ability and subsequent developmental competence of porcine oocytes. Oocytes were cultured in media containing 0, 300, 400 or 500 µg/ml of crocin. Upon maturation, the maturation rates, reactive oxygen species (ROS) and glutathione (GSH) levels, mRNA expression of genes (SOD, CAT, GPx, Bcl‐2, BAX and Caspase3), expression of cleaved caspase3 and subsequent embryo cleavage rates were measured. Results indicated that the maturation rate of the 400 µg/ml group was 86.80% (p < 0.01). The ROS concentration of the 500 µg/ml group was the lowest (p < 0.01). The GSH concentration of the 400 µg/ml group was the highest (p < 0.01). The SOD, CAT and GPx mRNA expression levels were the highest in the 300, 400 and 500 µg/ml groups, respectively, with the expression levels of all genes being significantly higher than that of the control group (p < 0.01). The Bcl‐2/BAX mRNA expression ratio in 400 and 500 µg/ml groups significantly higher than other groups and significantly decreased caspase3 expression level (p < 0.01). The expression level of cleaved caspase3 in the 500 µg/ml treatment group was the lowest, significantly lower than that of the control group (p < 0.01). The cleavage rate of the 400 µg/ml group was 62.50% (p < 0.01). These experimental results show that the supplementation of in vitro culture medium with 400 µg/ml of crocin significantly enhanced the antioxidant defence and anti‐apoptotic ability and subsequent cleavage rate of porcine embryo.     

Effects of dietary red yeast (Sporidiobolus pararoseus) on production performance and egg quality of laying hens

The purpose of this study was to evaluate the effect of dietary red yeast (Sporidiobolus pararoseus) on production performance and egg quality of laying hens. A total of 200 Esa Brown laying hens (23 weeks of age) were allocated equally to negative control group (no yeast supplement); positive control group (2 g/kg of Saccharomyces cerevisiae); 0.5, 1, 2 g/kg red yeast respectively. The experiment was lasted for 12 weeks. Feed intake, hen‐day egg production and egg weight were not different between control and supplemented groups. However, yeast‐supplemented groups were significantly improved feed efficiency (p < .05). Incremental levels of red yeast increased the colour score of egg yolk (p < .05). The cholesterol and triglyceride of serum and yolk were significantly (p < .05) lower in the laying hens fed dietary administration red yeast compared to the control diet; however, no significant (p > .05) differences among yeast‐supplemented groups were observed. The hepatic hydroxymethylglutaryl‐coenzymeA (HMG‐CoA) reductase activity was significantly lower (p < .05) in the 2 g/kg red yeast‐supplemented group compared to the control and other red yeast‐supplemented groups. Concentrations of caecal short‐chain fatty acids was increased (p < .05) in laying hens fed 1 and 2 g/kg red yeast as compared to the control group. Dietary administration of 2 g/kg red yeast (S. pararoceus) significantly improved egg yolk colour, decrease serum and egg yolk cholesterol levels.     

Effects of the E1 activating enzyme UBA2 on porcine oocyte maturation,apoptosis, and embryonic development in vitro

The purpose of this study was to investigate the effect of the E1 activating enzyme UBA2 on the expression of the SUMO-1 protein during in vitro maturation (IVM) of pig oocytes and embryonic development. In the 5 μg/ml UBA2 treatment group, the expression of the anti-apoptotic gene Bcl-2 and the embryo cleavage rate was significantly increased, while the proapoptotic gene Bax was significantly reduced. When 10 μg/ml UBA2 was added, the in vitro maturation rate, blastocyst rate, and SUMO-1 protein content of oocytes increased significantly (p < .05), and the expression of proapoptotic gene Caspase3 was significantly decreased (p < .05), while the viability of cumulus cells was extremely significantly reduced (p < .01). In summary, UBA2 can regulate the content of the SUMO-1 protein in mature pig oocytes in vitro, which in turn affects the maturation rate of oocytes, expression of apoptosis genes, cumulus cell viability, and the development of embryos after fertilization.     

Effects of berberine on the growth performance,antioxidative capacity and immune response to lipopolysaccharide challenge in broilers

This study evaluated the effects of berberine on growth performance, immunity, haematological parameters, antioxidant capacity, and the expression of immune response‐related genes in lipopolysaccharide (LPS)‐challenged broilers. We assigned 120 one‐day‐old male broilers (Ross 308) to two treatment groups; each group included two subgroups, each of which included six replicates of five birds per replicate. The experiment used a 2 × 2 factorial arrangement with berberine treatment (0 or 60 mg/kg dietary) and challenge status [injection of saline (9 g/L w/v) or LPS (1.5 mg/kg body weight)] as the main factors. On days 14, 16, 18 and 20, broilers were intraperitoneally injected with LPS or physiological saline. Blood and liver samples were collected on day 21. Dietary berberine supplementation significantly alleviated the compromised average daily gain and average daily feed intake (p < 0.05) caused by LPS. The LPS challenge led to increased lymphocyte and white blood cell (WBC) counts, malondialdehyde (serum and liver) content, and immunoglobulin G and M, tumour necrosis factor‐α (TNF‐α) and interleukin‐1β (IL‐1β) expression (p < 0.05) and significantly reduced serum total superoxide dismutase (T‐SOD) activity (p < 0.05). Dietary berberine significantly mitigated the LPS‐induced decreases in the mRNA expression of nuclear factor‐kappa B (NF‐κB), TNF‐α, IL‐1β, inducible nitrite synthase and cyclooxygenase‐2 (p < 0.05) in the liver. In conclusion, berberine supplementation has a positive effect on LPS challenge, which may be related to the increase in antioxidant enzyme activity and inhibition of both NF‐κB signalling and the expression of inflammatory mediators.     

Effects of chestnut tannins on intestinal morphology,barrier function,pro‐inflammatory cytokine expression,microflora and antioxidant capacity in heat‐stressed broilers

A study was conducted to evaluate the effects of chestnut tannins (CT) on intestinal morphology, barrier function, pro‐inflammatory cytokine expression, microflora and antioxidant capacity in heat‐stressed broilers. Four hundred 28‐day‐old male Ross 308 broilers were randomly assigned into four groups, with 10 replicates per group and 10 broilers per replicate. The broilers in the normal (NOR) group were kept at 22 ± 1°C and fed the basal diet, and each of the other three groups were treated with cyclic heat (33 ± 1°C from 0800 to 1800 and 22 ± 1°C from 1800 to 0800) and fed the basal diet with 0 (HT), 1 (CT1) or 2 (CT2) g of CT/kg of diet. The experiment lasted for 14 days. Compared with the HT group, broilers in the NOR and CT2 groups had higher (p < .05) average daily gain and villus height in the jejunum and lower serum d ‐lactate (p < .001) and diamine oxidase (p < .01) levels. The addition of 2 g CT/kg of diet increased the total antioxidant capacity (p < .001) and superoxide dismutase activities (p < .05) and zonula occludens‐1 mRNA expression level (p < .05) and decreased the malondialdehyde concentration (p < .01) and mRNA expression levels of interleukin‐6 (p < .001) and nuclear factor kappa B (p < .001) in the jejunal mucosa of heat‐stressed broilers. The populations of Escherichia coli and Clostridium in the jejunum (p < .01) and caecum (p < .05) of broilers in the HT group were higher than those in the NOR and CT2 groups. In conclusion, the addition of 2 g CT/kg of diet seemed to be a feasible means of alleviating the negative effects of heat stress on the growth performance and intestinal function of broilers.     

Use of polyvinyl alcohol as a chemically defined compound in egg yolk‐free extender for dog sperm cryopreservation

The objectives of this study were to investigate the effects of polyvinyl alcohol (PVA) as a chemically defined compound in egg yolk (EY)‐free extender by determining the appropriate concentration of PVA and the effect of pH adjustment in EY‐free PVA extenders on dog spermatozoa. Spermatozoa (1 × 10⁸ cells/ml) were frozen with EY‐free extenders supplemented with 0 (control), 0.025, 0.05, 0.1, 0.2 or 0.3 g/100 ml PVA. Sperm progressive motility (PM) was assessed immediately after thawing (IAT) and post‐thaw incubation (PTI), while viability, acrosome integrity and reactive oxygen species (ROS) levels were evaluated after PTI. Additionally, spermatozoa were frozen using EY‐free PVA extenders before pH adjustment (6.45) and after adjustment of pH (6.85). Viability, PM, ROS and gene expression (BCL2 and SMCP) were assessed. Supplementation with 0.05 g/100 ml or more PVA significantly increased PM compared to the control group in the IAT and PTI. Post‐thaw incubation significantly increased sperm motility in all groups. The acrosome integrity in all PVA groups was higher (p < .05) than the control without an effect on ROS and viability. Adjustment of the pH to 6.85 improved (p < .05) sperm PM compared to the non‐adjusted groups without affecting viability, ROS or expression of BCL2 and SMCP. We suggest that PVA supplementation in EY‐free Tris extenders can effectively protect dog spermatozoa during freezing and can maintain higher motility and acrosome integrity. Adjustment of pH in EY‐free PVA extenders can improve post‐thaw sperm motility. Therefore, PVA can be used as a compound in EY‐free extender for the cryopreservation of dog spermatozoa.     

Effects of Piper sarmentosum extract on the growth performance,antioxidant capability and immune response in weaned piglets

The biological properties of Piper sarmentosum render it a potential substitute for antibiotics in livestock feed. This study evaluated the effects of P. sarmentosum extract (PSE) on the growth performance, antioxidant capability and immune response of weaned piglets. Eighty 21‐d‐old weaned piglets were selected and randomly allocated to one of four dietary treatments with five replicates of four pigs each. The dietary treatments consisted of a basal diet supplemented with 0 (T0), 50 (T50), 100 (T100) or 200 (T200) mg/kg PSE. The feeding trial lasted 4 weeks. The results revealed that the T50 group had the highest average daily gain (ADG) and average daily feed intake (ADFI) throughout the feeding trial (p < 0.05). Additionally, the T50 group had higher (p < 0.05) serum glutathione peroxidase activity (GSH‐Px) and lower (p < 0.05) serum malondialdehyde (MDA) levels than the T0 group at 4 weeks post‐weaning (p < 0.05). Serum levels of interleukin‐1β (IL‐1β) and tumour necrosis factor‐α (TNF‐α) decreased, while serum levels of interleukin‐4 (IL‐4), interleukin‐10 (IL‐10) and transforming growth factor‐β (TGF‐β) increased by PSE supplementation at 4 weeks post‐weaning (p < 0.05). PSE supplementation upregulated the mRNA expression of IL‐4, IL‐10 and TGF‐β and downregulated the mRNA expression of TNF‐α, IL‐1β and interleukin‐6 (IL‐6) in the ileal mucosal layer of piglets (p < 0.05). In summary, our study findings revealed that PSE supplementation improved the antioxidant capability, and reduced inflammation, which may be beneficial to weaned piglet health.     

Montmorillonite improved the intestinal mucosal barrier functions of laying hens in late production

This study was conducted to investigate the effects of dietary supplementation with montmorillonite (MMT) on performance, intestinal endotoxin concentration, gut mucosal oxidation status, intestinal morphology and permeability, and immunological barrier function of laying hens during late production. Four hundred and eighty 75‐week‐old laying hens (Lohmann Brown) were randomly assigned to five treatments with eight replicates per treatment and 12 hens in each replicate. The hens were fed the basal diet supplemented with 0 (control), 0.3, 0.6, 0.9, or 1.2 g MMT/kg for 70 days. Compared with the control, supplemented with 0.9 g MMT/kg increased egg mass significantly (p < 0.05) during weeks 1–5 of the experiment. Supplemented with 0.6 and 0.9 g MMT/kg also increased the endotoxin concentration in the ileal digesta (p < 0.05), but decreased the MDA concentration in the ileum significantly (p < 0.05). The T‐AOC in the jejunum of the group fed 0.3 g MMT/kg was significantly increased (p < 0.05). Compared with the control, the villus height:crypt depth of ileum from the groups fed 0.6, 0.9, and 1.2 g MMT/kg increased significantly (p < 0.05). The sIgA concentration of jejunum in the groups fed 0.6 and 0.9 g MMT/kg was higher (p < 0.05) than the control. The MMT supplementation linearly increased (p < 0.05) the mRNA expression of claudin‐1 and claudin‐5 in the jejunum. Dietary MMT supplementation down‐regulated the mRNA expression of NF‐κB P65 and TNF‐α in the jejunum in a linear and quadratic manner (p < 0.05). The IL‐1β mRNA expression of jejunum in the group fed 0.6 g MMT/kg was lower (p < 0.05) than the control. In conclusion, dietary supplementation with MMT may improve the gut barrier functions and suggests that 0.9 g/kg of MMT in diets may be the optimal supplemental level for laying hens in late production.     

The effect of select seminal plasma proteins on endometrial mRNA cytokine expression in mares susceptible to persistent mating‐induced endometritis

In the horse, breeding induces a transient endometrial inflammation. A subset of mares are unable to resolve this inflammation, and they are considered susceptible to persistent mating‐induced endometritis PMIE Select seminal plasma proteins cysteine‐rich secretory protein‐3 (CRISP‐3) and lactoferrin have been shown to affect the innate immune response to sperm in vitro. The objective of this study was to determine whether the addition of CRISP‐3 and lactoferrin at the time of insemination had an effect on the mRNA expression of endometrial cytokines in susceptible mares after breeding. Six mares classified as susceptible to PMIE were inseminated during four consecutive oestrous cycles with treatments in randomized order of: 1 mg/ml CRISP‐3, 150 μg/ml lactoferrin, seminal plasma (positive control) or lactated Ringer's solution (LRS; negative control) to a total volume of 10 ml combined with 1 × 10⁹ spermatozoa pooled from two stallions. Six hours after treatment, an endometrial biopsy was obtained for qPCR analysis of selected genes associated with inflammation (pro‐inflammatory cytokines interleukin (IL)‐1β, IL‐8, tumour necrosis factor (TNF)‐α, interferon (INF)‐γ, anti‐inflammatory cytokines IL‐1RN and IL‐10, and inflammatory‐modulating cytokine IL‐6). Seminal plasma treatment increased the mRNA expression of IL‐1β (p = .019) and IL‐8 (p = .0068), while suppressing the mRNA expression of TNF (p = .0013). Lactoferrin also suppressed the mRNA expression of TNF (p = .0013). In conclusion, exogenous lactoferrin may be considered as one modulator of the complex series of events resulting in the poorly regulated pro‐inflammatory response seen in susceptible mares.     

Effects of supplemental conjugated linoleic acids (CLA) on fresh and post‐thaw sperm quality of Holstein bulls

This study was designed to investigate the effects of feeding‐protected conjugated linoleic acid (CLA) on the semen production and sperm freezability in Holstein bulls. Twelve bulls were randomly assigned to two groups (n = 6 per group). Bulls received the normal diet (control group) or the normal diet top‐dressed with 50 g of CLA (treated group) for 10 weeks. The control group received 40 g/day calcium soap of fatty acid. Fresh and post‐thaw semen quality was assessed on ejaculates collected at the 0, 4, 6, 8 and 10 week of supplementation. Semen evaluations including sperm concentration, motion characteristics (subjective and computer‐assisted), viability (Eosin–Nigrosin), membrane integrity (hypo‐osmotic swelling test) and abnormality were conducted. Semen volume, sperm concentration and total sperm output were not affected by dietary treatment (p > .05). The proportion of spermatozoa with abnormal morphology in fresh semen significantly increased (p < .05) in the CLA‐fed group compared to control group. Also, in CLA‐fed group, the proportion of post‐thaw spermatozoa with abnormal morphology at week 10 of trial was significantly higher in CLA than control group (p < .05). Progressive motility tended to be increased in the CLA‐fed group, although dietary supplementation did not affect other CASA parameters or viability in fresh and frozen‐thawed sperm. In this study, CLA supplementation had little positive effect on fresh or post‐thaw sperm quality of Holstein bulls.