Effects of xylo-oligosaccharides on growth，immunity and intestinal flora of Litopenaeus vannamei under low salinity stress
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摘要: 【目的】探究低盐胁迫下低聚木糖（XOS）对凡纳滨对虾（Litopenaeus vannamei）生长性能、抗氧化能力、免疫相关基因表达及肠道菌群结构的影响，为凡纳滨对虾淡化养殖饲料的开发提供理论依据。【方法】将450尾凡纳滨对虾按照养殖水体盐度和XOS添加量分为海水对照组HS0（盐度30‰+0 mg/kg XOS）及低盐对照组LS0（盐度3‰+0 mg/kg XOS）、LS250 （盐度3‰+250 mg/kg XOS）、LS500 （盐度3‰+500 mg/kg XOS）和LS1000 （盐度3‰+1000 mg/kgXOS）。饲养8周后计算生长指标，采用试剂盒测定消化酶活性和抗氧化指标，并通过实时荧光定量PCR检测免疫相关基因的表达情况。【结果】与HS0组相比，LS0组凡纳滨对虾的生长指标、消化酶活性、抗氧化能力及免疫相关基因相对表达量均有所下降。与LS0组相比，LS500和LS1000组凡纳滨对虾的增重率、特定生长率、存活率及淀粉酶（AMS）活性均呈显著升高趋势（P<0.05，下同），丙二醛（MDA）含量则呈显著降低趋势；脂肪酶（LPS）、胰蛋白酶（Trp）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性呈上升趋势，但差异不显著（P>0.05）。LS500和LS1000组凡纳滨对虾的酚氧化酶原（proPO）、抗脂多糖因子（ALF）、溶菌酶（LZM）、Toll样受体1（TLR1）、Toll样受体2（TLR2）和Toll样受体3（TLR3）基因的相对表达量均显著高于LS0组凡纳滨对虾。此外，LS1000组凡纳滨对虾的肠道菌群Shannon指数、Chao1指数和ACE指数显著高于LS0组凡纳滨对虾，Simpson指数显著低于LS0组凡纳滨对虾；在门分类水平上，变形菌门和拟杆菌门相对丰度显著降低，而蓝细菌门、厚壁菌门和疣微菌门的相对丰度显著增高；在属分类水平上，弧菌属、黄海绵菌属、斯氏弓型菌属和希瓦氏菌属相对丰度显著降低，而乳酸杆菌属和不动杆菌属相对丰度显著增加。【结论】在基础饲料中添加500~1000 mg/kg XOS，可显著提高凡纳滨对虾生长速率、抗氧化性能、免疫相关基因表达及改善肠道菌群组成结构，有效提高对虾在低盐胁迫下的养殖效果，即XOS可作为凡纳滨对虾淡化养殖饲料添加剂进一步开发应用。Abstract: 【Objective】 This study investigated the effects of xylo-oligosaccharide（XOS）on the growth performance， antioxidant capacity，immune-related gene expression and intestinal flora structure of Litopenaeus vannamei under low salinity stress conditions，and provided theoretical basis for the development of desalination culture feeds for L. vannamei. 【Method】 According to the salinity of the cultured water and XOS addition， the 450 L. vannamei were divided into seawater control group HS0（salinity 30‰+0 mg/kg XOS），low salinity control groups LS0（salinity 3‰+0 mg/kg XOS），LS250 （salinity 3‰+250 mg/kg XOS），LS500（salinity 3‰+500 mg/kg XOS）and LS1000（salinity 3‰+1000 mg/kg XOS）. Growth indexes were calculated after 8 weeks of feeding，digestive enzyme activities and antioxidant indexes were measured by kits，meanwhile，immune-related gene expression was detected by real-time fluorescence quantitative PCR. 【Result】 Compared with the HS0 group，the growth indexes，digestive enzyme activities，antioxidant capacity and the relative expression of immune-related genes of L. vannamei in the LS0 group were all decreased. Compared with the LS0 group，the weight gain rate，specific growth rate，survival rate and amylase（AMS）activity of L. vannamei in the LS500 and LS1000 groups showed significant increase trend（P<0.05，the same below），while the malondialdehyde（MDA） content showed a significant decrease trend. Lipase（LPS），trypsin（Trp），superoxide dismutase（SOD）and catalase （CAT）activities also showed an increasing trend，but the difference was not significant（P>0.05）. In LS500 and LS1000 groups，the relative expression levels of prophenoloxidase（proPO），anti-lipopolysaccharide factor（ALF），lysozyme （LZM），toll-like receptor 1（TLR1），toll-like receptor 2（TLR2）and toll-like receptor 3（TLR3）genes in LS500 and LS1000 groups were significantly higher than LS0. The intestinal flora Shannon index，Chao1 index，ACE index of LS1000 group L. vannamei were significantly higher than LS0 group， and Simpson index was significantly lower than LS0 group. At the phylum level，the relative abundance of Proteobacteria and Bacteroidetes decreased significantly， while the relative abundance of Cyanobacteria_Chloroplast，Firmicutes and Verrucomicrobia increased significantly. At the genus level，the relative abundance of Vibrio，Spongiimonas，Arcobacter and Shewanella decreased significantly， while the relative abundance of Lactobacillus and Acinetobacter increased significantly.【Conclusion】 Under low salinity stress，the addition of 500-1000 mg/kg XOS to the feed in this study significantly increased the growth rate，antioxidant performance，immune-related gene expression and improved the composition structure of intestinal flora of L. vannamei， effectively improving the culture effect of shrimp under low salitiny stress，so that XOS can be further developed and applied as a feed additive for desalination culture of L. vannamei.
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