Zhang Dong-Xing, Kang Yuan-Huan, Zhan Sheng, Zhao Ze-Lin, Jin Sheng-Nan, Chen Chong, Zhang Lei, Shen Jin-Yu, Wang Chun-Feng, Wang Gui-Qin, Shan Xiao-Feng, Qian Ai-Dong
College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Fish Health and Nutrition of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou, China.
Front Microbiol. 2019 Nov 15;10:2663. doi: 10.3389/fmicb.2019.02663. eCollection 2019.
is an emerging aquatic pathogen causing hemorrhagic septicemia in humans and animals. Probiotic is an effective strategy for controlling enteric infections through reducing intestinal colonization by pathogens. Here we report that the consumption of regulated the intestinal innate immune response and decreased the degree of intestinal inflammation damage caused by the in Crucian carp. In this study, we isolated four strains of , named C-11, S-22, L-17 and S-14 from apparently healthy Crucian carp, which exerted a broad-spectrum antimicrobial activity inhibiting both Gram-positive and Gram-negative bacteria especially the fish pathogens. isolates showed typical characteristics by endospore staining, physiological and biochemical test, enzyme activity analysis (amylase, protease, and lipase), and molecular identification. Here, -containing dietary was orally administrated to Crucian carp for 8 weeks before challenge. Immunological parameters and the expression of immune-related genes were measured at 2, 4, 6, 8, and 10 weeks post-administration. The results showed that was found to promote the increase in the phagocytic activities of peripheral blood leukocytes (PBLs) and head kidney leukocytes (HKLs), as well as the increase in interleukin 1β (IL-1β), IL-10 and tumor necrosis factor α (TNF-α) concentration of serum. Lysozyme levels (113.76 U/mL), ACP activity (25.32 U/mL), AKP activity (130.08 U/mL), and SOD activity (240.63 U/mL) were maximum ( < 0.05) in the C-11 treated group at 8 week. Our results showed that Crucian carp fed with the diet containing C-11 and S-22 developed a strong immune response with significantly higher ( < 0.05) levels of IgM in samples of serum, mucus of skin and intestine compared to L-17 and S-14 groups. Moreover, spores appeared to show no toxicity and damage in fish, which could inhabit the gut of Crucian carp. restrained up-regulation of pro-inflammation cytokines (IL-1β, IFN-γ, and TNF-α) mRNA levels in the intestine and head kidney at final stage of administration, and the expression of IL-10 was increased throughout the 10-week trial. infection increased the population of inflammatory cells in the intestinal villi in the controls. In contrast, numerous goblet cells and few inflammatory cells infiltrated the mucosa in the groups after challenge with . Compared with group, could safeguard the integrity of intestinal villi. The highest post-challenge survival rate (75.0%) was recorded in C-11 group. The present data suggest that probiotic act as a potential gut-targeted therapy regimens to protecting fish from pathogenic bacteria infection.
In this work, four strains isolated from apparently healthy Crucian carp, which exhibited a broad-spectrum antibacterial activity especially the fish pathogens. Administration of induced the enhancement of the intestinal innate immune response through reducing intestinal colonization by pathogens. The isolation and characterization would help better understand probiotic can be recognized as an alternative of antimicrobial drugs protecting human and animal health.
是一种新兴的水生病原体,可导致人类和动物患出血性败血症。益生菌是通过减少病原体在肠道的定植来控制肠道感染的有效策略。在此,我们报告食用 可调节鲫鱼肠道固有免疫反应,并降低由 引起的肠道炎症损伤程度。在本研究中,我们从看似健康的鲫鱼中分离出四株 ,分别命名为C-11、S-22、L-17和S-14,它们具有广谱抗菌活性,可抑制革兰氏阳性菌和革兰氏阴性菌,尤其是鱼类病原体。通过芽孢染色、生理生化试验、酶活性分析(淀粉酶、蛋白酶和脂肪酶)和分子鉴定, 分离株显示出典型的 特征。在此,在 攻击前,将含 的饲料口服给予鲫鱼8周。在给药后第2、4、6、8和10周测量免疫参数和免疫相关基因的表达。结果表明, 可促进外周血白细胞(PBLs)和头肾白细胞(HKLs)吞噬活性的增加,以及血清中白细胞介素1β(IL-1β)、IL-10和肿瘤坏死因子α(TNF-α)浓度的增加。在第8周时,C-11处理组的溶菌酶水平(113.76 U/mL)、酸性磷酸酶活性(25.32 U/mL)、碱性磷酸酶活性(130.08 U/mL)和超氧化物歧化酶活性(240.63 U/mL)最高(<0.05)。我们的结果表明,与L-17和S-14组相比,饲喂含C-11和S-22饲料的鲫鱼产生了强烈的免疫反应,血清、皮肤黏液和肠道样本中的IgM水平显著更高(<0.05)。此外, 芽孢在鱼类中似乎没有毒性和损害,可定殖于鲫鱼肠道。在给药末期, 抑制了肠道和头肾中促炎细胞因子(IL-1β、IFN-γ和TNF-α)mRNA水平的上调,并且在整个10周试验中IL-10的表达增加。在对照组中, 感染增加了肠绒毛中炎性细胞的数量。相比之下,在 攻击后, 组的黏膜中有大量杯状细胞且炎性细胞很少浸润。与 组相比, 可保护肠绒毛的完整性。C-11组在攻击后的存活率最高(75.0%)。目前的数据表明,益生菌 可作为一种潜在的肠道靶向治疗方案,保护鱼类免受病原菌感染。
在这项工作中,从看似健康的鲫鱼中分离出四株 菌株,它们具有广谱抗菌活性,尤其是对鱼类病原体。施用 可通过减少病原体在肠道的定植来诱导肠道固有免疫反应增强。该分离和鉴定将有助于更好地理解益生菌可被视为保护人类和动物健康的抗菌药物的替代品。