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一株潜在益生菌S12的分离及其对凡纳滨对虾生长性能、非特异性免疫力和抗病力的影响

Isolation of a putative probiotic strain S12 and its effect on growth performance, non-specific immunity and disease-resistance of white shrimp, Litopenaeus vannamei.

作者信息

Liu Hongyu, Li Zheng, Tan Beiping, Lao Ye, Duan Zhiyong, Sun Wuwei, Dong Xiaohui

机构信息

Laboratory of Aquatic Animal Nutrition and Feed, Fisheries College, Guangdong Ocean University, Zhanjiang 524025, PR China.

Kemin Industries (Zhuhai) Co. Ltd, Zhuhai 519110, PR China.

出版信息

Fish Shellfish Immunol. 2014 Dec;41(2):300-7. doi: 10.1016/j.fsi.2014.08.028. Epub 2014 Sep 16.

Abstract

The common pathogens in aquaculture are very different from those in terrestrial animals. The objective of this study was to isolate probiotic strain (s) from the digestive tract of healthy white shrimp Litopenaeus vannamei which was effective against aquatic animal pathogens. The putative probiotic strain S12 was identified as Bacillus subtilis based on the morphological and biochemical properties and 16S rDNA gene sequencing. The L. vannamei were fed with five different diets: control (basal diet with no probiotics or antibiotics), antibiotic control (basal diet supplemented with 0.3% florfenicol), basal diet supplemented with 5 × 10(9) cfu kg(-1) , 5 × 10(10) cfu kg(-1) and 5 × 10(11) cfu kg(-1) probiotic S12 (PS1-3). Each diet was randomly fed to quadruplication groups of 40 shrimps (0.4 ± 0.01 g) reared in tanks. After an 8-week feeding, the survival rate of shrimps fed with PS1 and PS3 were the highest among all treatments (P < 0.05). The moisture content of shrimps fed with florfenicol was significantly lower than that of the control group (P < 0.05). The supplement of probiotic S12 decreased the body crude lipid significantly (P < 0.05). The activities of phagocytic rate, lysozyme (LZ), superoxide dismutase phenoloxidase (SOD) and antibacterial activity were significantly higher than those in the control (P < 0.05), and the activities of SOD and the antibacterial activity in PS2 and PS3 were significantly higher than those in antibiotic control (P < 0.05). When infected with Vibrio harveyi at 4-weeks, the mortality was significantly lower (P < 0.05) in PS2 and PS3 groups than that in the control. After being infected with V. harveyi at 8-weeks, the mortality was significantly lower in the probiotic and antibiotic groups than that in the control (P < 0.05). This study suggested that probiotics could be used as an effective immunopotentiator, the optimal dose of the probiotic strain S12 is 5 × 10(10) cfu kg(-1) diet.

摘要

水产养殖中的常见病原体与陆生动物的有很大不同。本研究的目的是从健康凡纳滨对虾的消化道中分离出对水产动物病原体有效的益生菌菌株。根据形态学、生化特性和16S rDNA基因测序,推定的益生菌菌株S12被鉴定为枯草芽孢杆菌。给凡纳滨对虾投喂五种不同的饲料:对照组(不含益生菌或抗生素的基础饲料)、抗生素对照组(添加0.3%氟苯尼考的基础饲料)、添加5×10⁹ cfu kg⁻¹、5×10¹⁰ cfu kg⁻¹和5×10¹¹ cfu kg⁻¹益生菌S12的基础饲料(PS1 - 3)。每种饲料随机投喂到饲养在水箱中的40尾对虾(0.4±0.01 g)的四个重复组中。经过8周的投喂后,投喂PS1和PS3的对虾存活率在所有处理中最高(P<0.05)。投喂氟苯尼考的对虾的水分含量显著低于对照组(P<0.05)。添加益生菌S12显著降低了虾体粗脂肪含量(P<0.05)。吞噬率、溶菌酶(LZ)、超氧化物歧化酶、酚氧化酶(SOD)和抗菌活性显著高于对照组(P<0.05),并且PS2和PS3中的SOD活性和抗菌活性显著高于抗生素对照组(P<0.05)。在4周时感染哈维氏弧菌后,PS2和PS3组的死亡率显著低于对照组(P<0.05)。在8周时感染哈维氏弧菌后,益生菌组和抗生素组的死亡率显著低于对照组(P<0.05)。本研究表明益生菌可作为有效的免疫增强剂,益生菌菌株S12的最佳剂量为5×10¹⁰ cfu kg⁻¹饲料。

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