Effects of Hermetia illucens larvae meal on the Pacific white shrimp (Litopenaeus vannamei) revealed by innate immunity and 16S rRNA gene sequencing analysis.

Yongkang Chen, Zhenxiao Zhuang, Jieping Liu, Ziqiao Wang, Yucai Guo, Anqi Chen, Baoyang Chen, Wei Zhao, J. Niu
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Abstract

The larvae of the black soldier fly, Hermetia illucens, are now attracting attention and becoming promising sources for aquafeed ingredient due to the nutritious substance. However, the introduction of a novel ingredient into the recipe may have unpredictable effects on the innate immune function and gut bacteria composition of crustaceans. Therefore, the present study aimed to evaluate how dietary black soldier fly larvae meal (BSFLM) affected the antioxidant ability, innate immunity and gut microbiome of shrimp (Litopenaeus vannamei) fed with a practical diet, including the gene expression of Toll and immunodeficiency (IMD) pathways. Six experimental diets were formulated by replacing gradient levels of fish meal (0 %, 10 %, 20 %, 30 %, 40 % and 50 %) based on a commercial shrimp diet. Four replicates of shrimp were fed different diets three times daily for 60 days. Growth performance linearly decreased with increasing BSFLM inclusion. Results of antioxidative enzyme activities and gene expression suggested that low dietary BSFLM levels activated the antioxidant capacity of shrimp, while dietary BSFLM levels up to 100 g/kg may induce oxidative stress and inhibit glutathione peroxidase activity. Although traf6, toll1, dorsal and relish were significantly upregulated in different BSFLM groups, the expression of tak1 was significantly downregulated in groups containing BSFLM, implying the immune susceptibility may be weakened. Gut flora analysis indicated dietary BSFLM altered both beneficial and opportunistic pathogenic bacterial abundance, with low levels of dietary BSFLM increased the abundance of bacteria that may contribute to carbohydrate utilization, while high levels of dietary BSFLM may cause intestinal disease and low intestinal immune response. To conclude, 60-80 g/kg of dietary BSFLM showed no adverse effects on the growth, antioxidant capacity and gut flora of shrimp, which was the adequate level in shrimp diet. While 100 g/kg dietary BSFLM may induce oxidative stress and potentially weaken the innate immunity of shrimp.
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通过免疫和16S rRNA基因测序分析,揭示了黄颡鱼幼鱼饵料对凡纳滨对虾(Litopenaeus vanamei)的影响。
黑兵蝇(Hermetia illucens)的幼虫因其丰富的营养成分而备受关注,成为水产饲料原料的重要来源。然而,在配方中引入一种新成分可能对甲壳类动物的先天免疫功能和肠道细菌组成产生不可预测的影响。因此,本研究旨在评价饲粮中添加黑兵蝇幼虫粉(BSFLM)对凡纳滨对虾(Litopenaeus vannamei)抗氧化能力、先天免疫和肠道微生物组的影响,包括Toll和免疫缺陷(IMD)途径的基因表达。在商业对虾饲料的基础上,用梯度水平的鱼粉(0%、10%、20%、30%、40%和50%)替代6种试验饲料。试验4个重复,每天投喂3次不同饲料,连续投喂60 d。随着BSFLM添加量的增加,生长性能呈线性下降。抗氧化酶活性和基因表达结果表明,低水平的BSFLM可激活对虾的抗氧化能力,而高达100 g/kg的BSFLM可诱导氧化应激,抑制谷胱甘肽过氧化物酶活性。虽然在不同BSFLM组中traf6、tol1、dorsal和savor的表达均显著上调,但tak1的表达在含有BSFLM组中显著下调,表明免疫易感性可能减弱。肠道菌群分析表明,饲料中BSFLM改变了有益菌和机会致病菌的丰度,低水平的BSFLM增加了可能有助于碳水化合物利用的细菌丰度,而高水平的BSFLM可能导致肠道疾病和肠道免疫反应低下。综上所述,饲料中添加60 ~ 80 g/kg的BSFLM对对虾的生长、抗氧化能力和肠道菌群无不良影响,在对虾饲料中为适宜水平。而饲料中添加100 g/kg的BSFLM可能诱导氧化应激,并可能削弱对虾的先天免疫。
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