The Protective Effects Of Lactulose And Hydrogen-Rich Water On The Intestinal Injury In Weaned Piglets Fed Fusarium Contaminated Maize

Contamination of food by Fusarium mycotoxins is a problem for agricultural and food industries worldwide.Myocotoxins contaminate a large variety of dietary materials consumed by humans and animals and are commonly present in the food chain.The gastrointestinal tract is the primary target organ of food toxins,and exposure to high levels of toxic substances consequentially damages intestinal functions and health and suppresses growth performance.Hydrogen,the lightest gas,has been confirmed with antioxidant,anti-apoptotic and anti-inflammatory effects,which can prevent serious intestinal diseases.Lactulose can be used in the hindgut by microbial metabolism,and exerts a variety of physiological functions that are beneficial to intestinal health.Importantly,microbial metabolism of lactulose provides the host with a large amount of endogenous hydrogen.In this study,weaned piglets and intestinal cell lines were used as subjects.We evaluated the effects of hydrogen-rich water and lactulose on the growth performance and intestinal physiologic functions and damage induced by Fusarium mycotoxins.We demonstrate the protective effects of hydrogen gas in pigs.The resutls of this dissertation study suggest that dietary supplementation of hydrogen and/or lactulose is a novel solution to alleviate the intestinal toxicity and growth performance suppression caused by Fusarium mycotoxins in swine production.1 Effects of lactulose and hydrogen-rich water on growth suppression and oxidative stress in piglets fed Fusarium contaminated maizeThis study was to investigate the roles of lactulose(LAC)and hydrogen-rich water(HRW)in growth suppression and oxidative stress in piglets fed Fusarium contaminated maize.A total of 24 female piglets were evenly assigned to 4 groups:negative control(NC)group,mycotoxin contaminated(MC)feed group,MC feed with LAC treatment(MC+LAC),and MC feed with HRW treatment(MC+HRW),respectively.Piglets in the NC group were fed uncontaminated control diet,while remaining piglets were fed Fusarium mycotoxin-contaminated diet.For the NC and MC groups,10 mL· kg-1 body weight(BW)of hydrogen-free water(HFW)was orally administrated to piglets twice daily;while in the MC+LAC and MC+HRW groups,piglets were treated with the same dose of LAC solution dissolved in HFW(500 mg·kg-1 BW)and HRW twice daily,respectively.One piglet was removed from the MC,MC+LAC,and MC+HRW groups fed with a mycotoxin contaminated diet due to refusal to food and diarrhea.Therefore,five independent replicates from each group were used in this study.The body weight was recorded every week,in addition,feed intake,fecal scoring and diarrhea rate of piglets were recorded every day during the whole period of the experiment.The blood hydrogen levels before and after 2 h hydrogen-free water/HRW administration were detected at day 21.The hydrogen levels in gastrointestinal tract after 30 min hydrogen-free water/HRW administration and serum oxidative biomarkers and antioxidant enzymes were detected at the end of the experiment.Results showed that both HRW and LAC treatments significantly attenuated the reduction of average daily gain(ADG)and average daily feed intake(ADFI)caused by Fusarium mycotoxins,in addition,they protected against the mycotoxin-contaminated diet-induced higher diarrhea rate.HRW treatment increased the hydrogen levels in blood,stomach and duodenum after 2 h or 30 min oral administration,respectively.LAC treatment increased the hydrogen levels in blood before and after oral administration,increased the hydrogen levels of caecum and colon after 30 min oral administration,respectively.Most importantly,oral administration of HRW and LAC attenuated the Fusarium mycotoxins-induced oxidative stress.In conclusion,oral administration of hydrogen-rich water or lactulose could both protect against the growth reduction and oxidative damage caused by Fusarium mycotoxins.2 Effects of lactulose and hydrogen-rich water on intestinal microbiota in piglets fed Fusarium contaminated maizeThis study was to investigate the roles of lactulose(LAC)and hydrogen-rich water(HRW)in intestinal microbiota in piglets fed Fusarium contaminated maize.This experimental design was the same as in the previous chapter.On d 25,short-chain fatty acids(SCFAs)profiles in the digesta of the foregut and hindgut samples were assayed.The populations of selected bacteria and denaturing gradient gel electrophoresis(DGGE)profiles of total bacteria and methanogenic Archaea were also evaluated.Our data showed that Fusarium mycotoxins shifted the SCFAs production and populations and communities of microbiota.Both LAC and HRW protected against the Fusarium contaminated maize-induced lower SCFAs production in the digesta of the colon and caecum.In addition,the DGGE profile results indicated that LAC and HRW might shift the pathways of hydrogen-utilization bacteria,and change the diversity of intestine microbiota.Moreover,LAC and HRW administrations reversed the Fusarium contaminated maize-induced changing of the populations of Escherichia coli and Bifidobacterium in ileum digesta and hydrogen-utilizing bacteria in colon digesta.3 Effects of lactulose and hydrogen-rich water on intestinal epithelium morphology in piglets fed Fusarium contaminated maizeThis study was to investigate the roles of lactulose(LAC)and hydrogen-rich water(HRW)in preventing intestinal epithelium morphology damage in piglets fed Fusarium contaminated maize.This experimental design was the same as in the previous chapter.On d 25,intestinal tissues were sampled for morphological examination,morphological scores of histological lesions,intestinal epithelium apoptosis as well as relative genes expression analysis.Our data showed that Fusarium mycotoxins induced lower villus height(P<0.05),ratio of villus height to crypt depth(P<0.05)and morphological scores of histological lesions(P<0.05)in small intestine,greater apoptosis index(P<0.05)and high mRNA expression related to apoptosis(P<0.05).However,oral administrations of LAC and HRW attenuated the change of intestinal morphology,increased morphological scores of histological lesions.In addition,they provided beneficial effects in reducing apoptosis and the genes expression related apoptosis of epithelium cells in small intestine.In conclusion,the results showed that either LAC or HRW treatment could attenuate intestinal epithelium morphology damage in piglets fed Fusarium contaminated maize.4 Effects of lactulose and hydrogen-rich water on intestinal barrier function in piglets fed Fusarium contaminated maizeThis study was to investigate the roles of lactulose(LAC)and hydrogen-rich water(HRW)in preventing intestinal barrier function destruction in piglets fed Fusarium contaminated maize.This experimental design was the same as in the previous chapter.On d 25,serum was collected and used for biochemical analysis.Intestinal mucosa samples were collected for relative gene and protein expression analyses.Our data showed that Fusarium mycotoxins induced higher serum diamine oxidase(DAO)activities(P<0.05),D-lactic acid levels(P<0.05),endotoxin status(P<0.05)and high mRNA expression related to tight junction(P<0.05)in small intestine.In addition,the distribution and down-regulation of claudin-3(CLDN3)protein in the small intestine was also observed.However,oral administrations of LAC and HRW attenuated intestinal leaking and irregular CLDN3 distribution and restored the expression and distribution of CLDN3 protein in the small intestinal mucosal layer.In conclusion,the results showed that either LAC or HRW treatment could attenuate intestinal barrier destruction in piglets fed Fusarium contaminated maize.5 The effect of hydrogen gas on oxidative damage and apoptosis in IPEC-J2 cells induced by deoxynivalenolThis study was to investigate the effect of hydrogen gas on oxidative damage and apoptosis in intestinal porcine epithelial cells(IPEC-J2 cells)induced by deoxynivalenol(DON).In the present study,IPEC-J2 cells were assigned to 4 groups:control group,5 ?M DON group(DON group),hydrogen-saturated medium group(H2 group),5 ?M DON and hydrogen-saturated medium(DON+H2 group).IPEC-J2 cells in four groups were subjected to the various treatments for 12 h.Then the cellular morphology was observed.Cell viability was determined by the MTT assay and the level of lactate dehydrogenase(LDH)in culture supernatant was measured.The concentrations of cellular catalase(CAT),total superoxide dismutase(T-SOD),malonaldehyde(MDA),8-hydroxy-2'-deoxyguanosine(8-OHdG)and 3-nitrotyrosine(3-NT)were also determined.In addition,the mRNA expression of genes related to antioxidant enzymes and apoptosis,and the Caspase-3 and Bax proteins expression were analyzed.Our data revealed that 5 ?M DON caused cells shedding,lowered cell viability(P<0.05),increased the LDH level in culture supernatant(P<0.05),decreased the activities of CAT and T-SOD(P<0.05);however,increased the levels of MDA,8-OHdG and 3-NT(P<0.05).The mRNA expressions of genes related to antioxidant enzymes were decreased(P<0.05).Moreover,DON treatment decreased the mRNA expression of Bcl-2 and increased the mRNA and protein expression of Caspase-3 and Bax(P<0.05).The use of hydrogen-saturated medium significantly attenuated oxidative stress and apoptosis in IPEC-J2 induced by DON.In conclusion,hydrogen gas protects IPEC-J2 from DON-induced oxidative damage and apoptosis.