| The performance,health,and well-being of livestock are strongly influenced by climate.Higher ambient temperature and humidity are environmental stressors that cause stress on animals.On the other hand,heat stress is the major factor in reducing livestock production,reproduction and immunity.Trace elements such as Selenium(Se)and Zinc(Zn)are essential for maintaining health and immunity and play an important role in antioxidant defense and immunity function and they are involved in growth,production and reproduction and act as cofactors of enzymes which are important to the immunity of animal.Moreover,probiotics are essentially living microorganisms that,when applied in sufficient quantities,give the host health benefits and have beneficial effects on host animals.Therefore,the purpose of this study was to assess that Selenium/Zinc-Enriched probiotics improve growth performance,antioxidant status,serum enzyme activity,immune Function,physiological function of the liver,related gene expression,and intestinal morphological characteristics of Wistar rats raised under high ambient temperature that will be present in the following experiments.Experiment 1:Zinc-Enriched Probiotics improve growth performance,antioxidant status,immune function,gene expression and morphological characteristics of Wistar rats raised under high ambient temperatureThe present study was conducted to evaluate the effects of Zinc-enriched probiotics(ZnP)on growth performance,antioxidant status,immune function,related gene expression and morphological characteristics of Wistar rats raised under high heat stress condition during summer.36,six-week-old male Wistar rats were randomly divided into three groups;fed with basal diet(control group),basal diet with probiotics(P)and basal diet with Zinc-enriched Probiotics supplementation(ZnP,100mg/L),for 40 consecutive days.Blood samples were collected through intra-cardiac method on the last day of experiment and tissues were collected from liver,heart and kidneys.The results revealed that both P and ZnP significantly(P<0.05)enhanced growth performance.However,ZnP remarkably increased Glutathione content,Glutathione peroxidase,and Superoxide dismutase activities but reduced malondialdehyde level in serum of the Wistar rats.The concentration of IL-2,IL-6,and IFN-y were significantly(P<0.05)increased with treatments of P and ZnP compared to control group while IL-10 was significantly(P<0.05)decreased.Additionally,the expression of SOD 1,SOD2,MT1,and MT2 genes were significantly(P<0.05)up-regulated with the treatment of ZnP,but Hsp90 and Hsp70 heat shock genes were significantly(P<0.05)down-regulated with the treatment of P and ZnP respectively.Hematoxylin and Eosin staining showed that both P and ZnP supplementation treatments induced changes in Villus height and Intestinal wall thickness.In conclusion,ZincEnriched Probiotics supplementation can improve the growth performance of Wistar rats under high ambient temperature through enhancing antioxidant status,immune function,related genes expression and intestinal morphological characteristics.This product may serves as a potential nutritive supplement for Wistar rats under high heat stress conditions.Experiment 2:Selenium/Zinc-Enriched probiotics improve serum enzyme activity,antioxidant ability,inflammatory factors and related gene expression of Wistar rats inflated under heat stressAims:The present study was carried out to investigate the influences of Selenium/Zinc-Enriched probiotics(SeZnP)on growth performance,serum enzyme activity,antioxidant capability,inflammatory factors and gene expression associated with Wistar rats inflated under high ambient thermal-stress.Main methods:Sixty male rates with six-weeks of age were randomly allocated into five groups(12 per group)and fed basal diet(Control group),basal diet supplemented with probiotics(P),Zinc-Enriched Probiotics(ZnP,100mg/L),Selenium-Enriched Probiotics(SeP,0.3mg/L)and Selenium/Zinc-Enriched Probiotics(SeZnP,0.3mg+100mg/L).The experiment lasted 30 days.Blood and Tissues samples were taken to investigate serum enzyme activity,antioxidants capability and inflammatory factors by using of commercial kits and antioxidant,heat shock and inflammatory related molecules expressions were determined by qRT-PCR.Key findings:Data analysis revealed that thermal stress significantly increased the level of Aspartate-aminotransferase,Alanine-aminotransferase,Lactate-dehydrogenase,Creatine-kinase,blood urea nitrogen,Creatinine and Alkaline phosphatase compared to P,ZnP,SeP or SeZnP groups(P<0.01).However,supplementation of ZnP,SeP,and SeZnP significantly enhanced glutathione content,glutathione-peroxidase&superoxide-dismutase activity,and decreased malondialdehyde content(P<0.05).Moreover,the concentration of IL-2,IL-6 and IL-8 were significantly increased while IL-10 was significantly decreased(P<0.05).Furthermore,the expression of GPx1 and SOD1 genes were significantly increased,but COX-2,iNOS,HSP70 and 90 mRNA levels were significantly decreased(P<0.05).Finally,the highest influence of the mentioned parameters was observed in SeZnP supplemented group.Significance:Our study suggests that SeZnP supplementation serves as possible and best nutritive than ZnP or SeP for Wistar rats raising under high ambient temperature.Experiment 3:Hepatoprotective effects of Selenium-Enriched probiotics supplementation on heat-stressed Wistar rat through anti-inflammatory and antioxidant effectsThe purpose of this study was to elucidate the effect of Selenium-Enriched probiotics on the liver of heat-treated Wistar rats.48,10-weeks-old male rats were grouped into control(Con),high temperature(HT),high temperature plus probiotics(HT+P),high temperature plus Selenium-Enriched probiotics groups(HT+SeP)and fed with selenium containing 0.3mg/kg,L.acidophilus 1011/mL and S.cerevisiae 10 9/mL.After keeping the rats on their respective diets for 42 days,the HT group,HT+P group,and HT+SeP group were exposed to higher ambient temperature(40-42℃),while control group were kept at room temperature(25 ℃).After 42 days of thermal exposure,blood and liver tissues were collected.The results indicate that body weight of rats in HT group decreased but liver weight and liver index were increased.Histological examination showed dilatation in the hepatic sinusoids,an interstitial venous congestion in HT group.Moreover,in the HT group,hepatic MDA content,serum AST,ALT,ALP and LDH levels were significantly increased,while hepatic SOD,T-AOC,GSH-PX activities and serum total protein level were significantly reduce as compared to control.The histomorphology of the liver in HT+P and HT+SeP groups were restored,and serum AST,ALT,ALP,LDH and hepatic MDA levels decreased significantly,but the serum total protein levels and SOD,T-AOC,and GSH-PX activities were increased significantly in the liver tissues as compare to HT group.In addition,the expression levels of mRNA Gpx1,SOD1,Nrf2,and Bcl-2 were significantly increased,while expression levels of Bax,IL-6,TNF-α,COX-2,NF-κB,αSMA,TGFβ1,Collagen Ⅰ,HSP70,and HSP90 were significantly decreased after SeP supplementation(P<0.01).From this study we conclude,that SeP can protect Wistar rat from oxidative stress,inflammation,apoptosis,and liver fibrosis induced by heat stress. |
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