Inhibitory activity of native probiotic Bacillus vallismortis IS03 against pathogenic Vibrio harveyi under in vitro and in vivo conditions in Litopenaeus vannamei

M. Mahjoub, M. Mirbakhsh, M. Afsharnasab, S. Kakoolaki, S. Hosseinzadeh
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Abstract

An excellent alternative for chemical antimicrobial agents to prevent disease in the shrimp aquaculture is the application of effective probiotics. The present study was evaluated the effect of Bacillus vallismortis IS03 as a native probiotic, isolated from digestive tract of Litopenaeus vannamei on pathogenic Vibrio harveyi under in vitro and in vivo circumstances. Co-cultivation of V. harveyi and B. vallismortis showed significantly (P<0.05) decreased the growth of V. harveyi in the treatment groups compared to the control. Cell-free extracts of B. vallismortis IS03 exhibited more appropriate antibacterial effects on replication of V. harveyi. The highest and lowest inhibitory effects were respectively shown in10 and 10 CFU ml of B. vallismortis IS03 cell-free extracts. The probiotic potential of B. vallismortis IS03 was assessed through the groups of control and the experiments 10, 10 and 10 CFU ml salt water once every 3 days from zoea1 process to end point of the study. Correspondence M. Mirbakhsh, Iranian Fisheries Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran (e-mail: maryam.mirbakhsh@gmail.com). The probiotic potential of B. vallismortis IS03 was assessed through the groups of control and the experiments 10, 10 and 10 CFU ml salt water once every 3 days from zoea1 process to end point of the study. Shrimp survival was determined after 10 days of challenge with V. harveyi at 10 5 CFU ml (for the first 5 days) and 10 7 CFU ml (for the second 5 days).The cumulative mortality in the treatment with 10 CFU ml of B. vallismortis IS03 reached 11.88% compared to 40.63% in the control group. At the end of the trial, total bacterial counts on TSA, total vibrio on TCBS were significantly (P<0.05) lower in the 10 CFU ml treatment group. Bacillus counts on MYP agar in the treatment groups were significantly (P<0.05) higher than the control, also total bacterial counts was lower in the treatment groups, while, no vibrio were grown in the muscle tissues of shrimp treated with probiotics. It is concluded that 10 CFU ml of probiotic, B. vallismortis IS03 has antibacterial efficiency against pathogenic V. harveyi at in vitro and in vivo conditions.
原生益生菌vallismortis IS03在体外和体内条件下对致病性哈威弧菌的抑制活性研究
有效益生菌的应用是化学抗菌剂在对虾养殖中预防疾病的一个很好的替代方案。本研究从凡纳滨对虾(Litopenaeus vallismortis IS03)消化道中分离得到的原生益生菌对致病性哈威氏弧菌(Vibrio harveyi)的体外和体内作用进行了研究。与对照组相比,harveyi弧菌与B. valismortis共培养各组harveyi弧菌的生长显著(P<0.05)降低。vallismortis IS03无细胞提取物对harveyi弧菌的复制表现出较好的抑菌作用。在10 CFU ml和10 CFU ml的无细胞提取物中,抑菌效果最高,抑菌效果最低。从zoea1过程到研究结束,通过对照组和试验组10、10和10 CFU ml盐水,每3天1次,对vallismortis IS03的益生菌潜力进行评估。伊朗德黑兰农业研究、教育和推广组织(AREEO)伊朗渔业科学研究所M. Mirbakhsh (e-mail: maryam.mirbakhsh@gmail.com)。从zoea1过程到研究结束,通过对照组和试验组10、10和10 CFU ml盐水,每3天1次,对vallismortis IS03的益生菌潜力进行评估。以105cfu ml(前5天)和107cfu ml(后5天)的哈维氏弧菌攻毒10天后测定虾的存活率。10 CFU ml的vallismortis IS03治疗组的累计死亡率为11.88%,对照组为40.63%。试验结束时,10 CFU ml治疗组TSA总细菌数、TCBS总弧菌数显著(P<0.05)降低。各处理组MYP琼脂上芽孢杆菌数量显著(P<0.05)高于对照组,总细菌数量低于对照组,而益生菌处理对虾肌肉组织中未生长弧菌。综上所述,10 CFU ml的益生菌B. vallismortis IS03在体外和体内条件下均对致病性哈维氏弧菌具有抑菌效果。
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