Grupo de Investigación en Acuicultura (GIA), Instituto Universitario Ecoaqua, Universidad de Las Palmas de Gran Canaria, Crta. Taliarte s/n, 35214 Telde, Canary Islands, Spain.
Grupo de Investigación en Acuicultura (GIA), Instituto Universitario Ecoaqua, Universidad de Las Palmas de Gran Canaria, Crta. Taliarte s/n, 35214 Telde, Canary Islands, Spain.
Fish Shellfish Immunol. 2018 Oct;81:10-20. doi: 10.1016/j.fsi.2018.07.007. Epub 2018 Jul 5.
The aim of this study was to assess the effects of dietary mannan oligosaccharides (MOS), Pediococcus acidilactici or their conjunction as a synbiotic in low fish meal (FM) and fish oil (FO) based diets on European sea bass (Dicentrarchus labrax) disease resistance and gut health. For that purpose, sea bass juveniles were fed one of 6 diets containing different combinations of MOS (Biomos and Actigen; Alltech, Inc., Kentucky, USA) and Pediococcus acidilactici (BAC, Bactocell; Lallemand Inc., Cardiff, UK) replacing standard carbohydrates as follows (MOS (%)/BAC (commercial recommendation): high prebiotic level (HP) = 0.6/0, low prebiotic level (LP) = 0.3/0, only probiotic (B) = 0/+, high prebiotic level plus probiotic (HPB) = 0.6/+, low prebiotic level plus probiotic (LPB) = 0.3/+, control (C) = 0/0 for 90 days. After 60 and 90 days of feeding trial, fish were subjected to an experimental infection against Vibrio anguillarum. Additionally, inducible nitric oxide synthase (iNOS) and tumor necrosis factor α (TNFα) gut patterns of immunopositivity and major histocompatibility complex class II (MHCII), transforming growth factor β (TGF-β), regulatory T-cell subset (CD4T lymphocytes) and effector T cell (CD8αT lymphocytes) gene expression patterns in gut by in situ hybridization were evaluated after 90 days of feeding. The effects of both additives on posterior gut through Gut Associated Lymphoid Tissue (GALT) gene expression was also studied. Fish fed the prebiotic and its combination with P. acidilactici presented increased weight regardless of the dose supplemented after 90 days of feeding, however no effect was detected on somatic indexes. For posterior gut, morphometric patterns and goblet cells density was not affected by MOS, P. acidilactici or its combination. Anti-iNOS and anti-TNFα gut immunopositivity patterns were mainly influenced by MOS supplementation and not by its combination with P. acidilactici. MHCII-β, TCR-β, CD4 and CD8-α positive cells distribution and incidence was not affected by diet. Fish fed HP dose presented a clear up-regulation of TNF-α, cyclooxygenase-2 (COX-2), CD4 and IL10, whereas P. acidilactici dietary supplementation increased the number of interleukin-1β (IL1β) and COX-2 gene transcripts. Synbiotic supplementation resulted in a reduction of MOS-induced gut humoral proinflammatory response by increasing the expression of some cellular-immune system related genes. Fish mortality after V. anguillarum infection was reduced in fish fed LPB and LP diets compared to fish fed the non-suppelmented diet after 90 days of feeding. Thus, overall pointing to the combination of a low dose of MOS and P. acidilactici as synbiont (LPB) as a viable tool to potentiate European sea bass juvenile's growth and disease resistance when supplemented in low FM and FO diets.
本研究旨在评估低鱼粉(FM)和鱼油(FO)基础日粮中添加甘露寡糖(MOS)、植物乳杆菌或其共生体作为合生元对欧洲鲈鱼(Dicentrarchus labrax)疾病抵抗力和肠道健康的影响。为此,将鲈鱼幼鱼分别投喂 6 种不同组合的 MOS(Biomos 和 Actigen;Alltech,Inc.,肯塔基州,美国)和植物乳杆菌(BAC,Bactocell;Lallemand Inc.,加拿大魁北克省)的日粮,用标准碳水化合物替代,具体组合如下(MOS(%)/BAC(商业推荐):高益生元水平(HP)=0.6/0,低益生元水平(LP)=0.3/0,仅益生菌(B)=+/0,高益生元水平加益生菌(HPB)=0.6/+,低益生元水平加益生菌(LPB)=0.3/+,对照组(C)=0/0,为期 90 天。在 60 和 90 天的喂养试验后,对鱼类进行鳗弧菌的实验性感染。此外,在 90 天的喂养后,通过原位杂交评估诱导型一氧化氮合酶(iNOS)和肿瘤坏死因子 α(TNFα)肠道免疫阳性模式以及主要组织相容性复合体 II 类(MHCII)、转化生长因子 β(TGF-β)、调节性 T 细胞亚群(CD4T 淋巴细胞)和效应 T 细胞(CD8αT 淋巴细胞)基因表达模式。还研究了两种添加剂对后肠通过肠道相关淋巴组织(GALT)基因表达的影响。无论补充剂量如何,在 90 天的喂养后,投喂前生物和其与植物乳杆菌组合的鱼类体重增加,但体脂指数没有变化。对于后肠,MOS、植物乳杆菌或其组合对肠道形态和杯状细胞密度没有影响。抗 iNOS 和抗 TNFα 肠道免疫阳性模式主要受 MOS 补充的影响,而不受其与植物乳杆菌组合的影响。MHCII-β、TCR-β、CD4 和 CD8-α阳性细胞的分布和发生率不受饮食的影响。投喂 HP 剂量的鱼类 TNF-α、环加氧酶-2(COX-2)、CD4 和 IL10 的表达明显上调,而植物乳杆菌的日粮补充增加了白细胞介素-1β(IL1β)和 COX-2 基因转录本的数量。共生体补充减少了 MOS 诱导的肠道体液性促炎反应,增加了一些与细胞免疫相关基因的表达。在 90 天的喂养后,与未补充饮食的鱼类相比,投喂 LPB 和 LP 日粮的鱼类在鳗弧菌感染后的死亡率降低。因此,总体而言,低剂量 MOS 和植物乳杆菌的组合(LPB)作为合生元,在低 FM 和 FO 日粮中添加时,是增强欧洲鲈鱼幼鱼生长和抗病能力的可行工具。