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投喂富含黑水虻幼虫(BSF)且添加几丁质酶的饲料后,尼罗罗非鱼在大肠杆菌脂多糖(LPS)刺激下的肠道组织病理学和免疫应答。

Intestinal histopathology and immune responses following Escherichia coli lipopolysaccharide challenge in Nile tilapia fed enriched black soldier fly larval (BSF) meal supplemented with chitinase.

机构信息

Research Unit in Environmental and Evolutionary Biology (URBE), Institute of Life, Earth and Environment (ILEE), University of Namur, Belgium; Laboratory of Hydrobiology and Aquaculture (LHA), Faculty of Agricultural Sciences, University of Abomey-Calavi, Cotonou, Benin.

Research Unit in Environmental and Evolutionary Biology (URBE), Institute of Life, Earth and Environment (ILEE), University of Namur, Belgium.

出版信息

Fish Shellfish Immunol. 2022 Sep;128:620-633. doi: 10.1016/j.fsi.2022.08.050. Epub 2022 Aug 28.

Abstract

This study aimed to determine to what extend the addition of chitinase to black soldier fly larvae (BSF) meals enriched with either PUFA or LC-PUFA could improve the gut health of Nile tilapia and increase its immune status. Two types of BSF meals enriched with either α-linolenic acid (ALA) or ALA + eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) were produced using BSF larvae cultured on vegetable substrates (VGS) or fish offal substrates (FOS), respectively. Seven diets were formulated: a control FMFO diet and two other control diets VGD0 vs FOD0 containing the meals of each type of BSF meal as total replacement for fishmeal (FM) and fish oil (FO), as well as four diets supplemented with chitinase. Two doses of commercial chitinase from Aspergillus niger (2 g/kg and 5 g/kg of feed) were supplemented to the other diets VGD0 and FOD0 to formulate VGD2, VGD5, FOD2 and FOD5. After 53 days of feeding, FOD5 diet induced a similar growth performance as the FMFO control diet, while a significant decrease of growth was observed for the other BSF larval-based diets. BSF/FOS meal led to higher SGR of fish than BSF/VGS, as for the FOD5 compared to VGD5. At day 53, lysozyme values showed an increasing trend in fish fed all the BSF-based diets, especially those fed the VGD5. After the Escherichia coli lipopolysaccharide (LPS) injection (day 54), the same increasing trend was observed in lysozyme activity, and modulation was observed only in the VGD5 fish. ACH50 activity was reduced by the BSF-based diets except for the FOD5 diet at day 53, and LPS modulation was only observed for the VGS-chitinase-based diets at day 54. Peroxidase activity and total immunoglobulin (Igs) blood level were not affected by substrate, chitinase dose or LPS injection. At day 53, the low or high dose of chitinase increased the expressions of tlr2, il-1β and il-6 genes in the head kidney of fish fed the BSF/VGS diets compared to those fed the VGD0 or FMFO control diets. At day 54 after LPS injection, the high dose of chitinase decreased the expressions of tlr5 gene in the spleen and mhcII-α gene in the head kidney of fish fed FOD5 diets compared to those fed FOD0 diets. BSF/VGS but not BSF/FOS based diets increased fish sub-epithelial mucosa (SM) and lamina propria (LP) thickness and the number of goblet cells (GC) in fish, but dietary chitinase seemed to prevent some of these effects, especially at low dose. Results showed that chitinase supplementation of 5 g/kg of chitinase to a BSF-based diet enriched with LC-PUFA improved growth, prevented histological changes in the proximal intestine and enhanced some innate immune functions of Nile tilapia without any clear booster effect after challenge with E. coli LPS.

摘要

本研究旨在确定在富含多不饱和脂肪酸(PUFA)或长链多不饱和脂肪酸(LC-PUFA)的黑水虻幼虫(BSF)饲料中添加几丁质酶能在何种程度上改善尼罗罗非鱼的肠道健康并提高其免疫状态。使用分别以蔬菜底物(VGS)或鱼内脏底物(FOS)培养的 BSF 幼虫生产了两种富含 α-亚麻酸(ALA)或 ALA+二十碳五烯酸(EPA)+二十二碳六烯酸(DHA)的 BSF 饲料。设计了七种饲料:对照 FMFO 饲料和两种对照饲料 VGD0 与 FOD0,其中每种 BSF 饲料的饲料均全部替代鱼粉(FM)和鱼油(FO),以及四种添加几丁质酶的饲料。在其他两种对照饲料 VGD0 和 FOD0 中添加了两种剂量的商业黑曲霉几丁质酶(2g/kg 和 5g/kg 的饲料),以分别制成 VGD2、VDG5、FOD2 和 FOD5。在 53 天的喂养后,FOD5 饲料诱导的生长性能与 FMFO 对照饲料相似,而其他基于 BSF 幼虫的饲料则观察到生长显著下降。BSF/FOS 饲料导致鱼类的特定生长率(SGR)高于 BSF/VGS,FOD5 饲料也高于 VGD5 饲料。在第 53 天,溶菌酶值在所有基于 BSF 的饲料喂养的鱼类中呈上升趋势,尤其是在 VGD5 饲料喂养的鱼类中。在注射大肠杆菌脂多糖(LPS)(第 54 天)后,溶菌酶活性也观察到相同的上升趋势,并且仅在 VGD5 鱼类中观察到调制。ACH50 活性除 FOD5 饲料外,在第 53 天受到基于 BSF 的饲料的抑制,而 LPS 调制仅在第 54 天在基于 VGS 的几丁质酶饲料中观察到。过氧化物酶活性和总免疫球蛋白(Igs)血液水平不受底物、几丁质酶剂量或 LPS 注射的影响。在第 53 天,低或高剂量的几丁质酶增加了与 VGD0 或 FMFO 对照饲料相比,喂食 BSF/VGS 饲料的鱼类头肾中 tlr2、il-1β 和 il-6 基因的表达。在 LPS 注射后第 54 天,高剂量的几丁质酶降低了喂食 FOD5 饲料的鱼类脾脏中的 tlr5 基因和头肾中的 MHCII-α 基因的表达与喂食 FOD0 饲料的鱼类相比。BSF/VGS 饲料而不是 BSF/FOS 饲料增加了鱼类亚上皮黏膜(SM)和固有层(LP)的厚度和杯状细胞(GC)的数量,但日粮几丁质酶似乎预防了其中一些变化,尤其是低剂量时。结果表明,在富含 LC-PUFA 的 BSF 饲料中添加 5g/kg 的几丁质酶可改善生长,防止近端肠道的组织学变化,并增强尼罗罗非鱼的某些先天免疫功能,而在大肠杆菌 LPS 攻毒后没有明显的增强作用。

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