THE EFFECT OF Bacillus sp. ADDITION ON THE VIRULENCE OF Vibrio parahemolyticus TOWARD AXENIC CULTURE OF Artemia franciscana

Ni Luh Komang Ayu Maitri Jayanthi, Pande Gde Sasmita Julyantoro, D. Pebriani
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Abstract

Research on the benefits of Bacillus sp. in aquaculture has been done a lot. Currently, the aquaculture industry, especially shrimp farming, is facing AHPND disease, which is known to be caused by Vibrio parahaemolyticus. This study aims to see the effect of Bacillus sp. against virulence of V. parahaemolyticus in vitro using caseinase production as an indicator and survival of Artemia franciscana cultured axenically when challenged with V. parahaemolyticus. In in vitro tests, cultures of V. parahaemolyticus were grown with and without the addition of Bacillus sp. on Luria Bertani (LB) liquid media, and then the caseinase test was carried out using skim milk (SM) agar as a growth medium. In the challenge test, 10 axenic cultured Artemia sp. were distributed in falcon tubes containing 10 ml of sterile seawater. Challenge test treatments consisted of treatment A as control of Artemia culture without the addition of bacteria, treatment B Artemia culture adding by 106 CFU/ml V. parahaemolyticus, treatment C Artemia culture with the addition of 106 CFU/ml Bacillus sp., and treatment D Artemia culture with the addition of 106 CFU/ml V. parahaemolyticus and 106 CFU/ml Bacillus sp. All treatments were done in triplicates. The results showed that the addition of Bacillus sp. was able to reduce the caseinase production of V. parahemolyticus up to 29% indicated by the decrease  of the clearing zone diameter formed on SM agar. Furthermore, the addition of Bacillus sp. in treatment D was able to significantly increasing (P<0.05) of Artemia's survival when challenged with V. parahamolyticus. This indicates that Bacillus sp. has a potential as a probiotic candidate to prevent disease caused by V. parahaemolyticus.
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添加芽孢杆菌对青蒿无菌培养副溶血性弧菌毒力的影响
人们对芽孢杆菌在水产养殖中的益处进行了大量的研究。目前,水产养殖业,特别是对虾养殖业正面临着AHPND疾病,这种疾病已知是由副溶血性弧菌引起的。本研究旨在观察芽孢杆菌对副溶血性弧菌体外毒力的影响,以酪蛋白酶产量为指标,以及无菌培养的对副溶血性弧菌的拮抗作用。在体外试验中,分别在贝塔尼Luria Bertani (LB)液体培养基上添加芽孢杆菌和不添加芽孢杆菌培养副溶血性弧菌,然后以脱脂乳(SM)琼脂为培养基进行酪蛋白酶试验。攻毒试验中,将10只无菌培养的Artemia sp分布在含有10 ml无菌海水的猎鹰管中。攻毒试验处理为:A组为不添加细菌的对照组,B组为添加106 CFU/ml副溶血性弧菌的对照组,C组为添加106 CFU/ml副溶血性弧菌的对照组,D组为添加106 CFU/ml副溶血性弧菌和106 CFU/ml芽孢杆菌的对照组。结果表明,芽孢杆菌的添加可使副溶血性弧菌的干酪酶产量降低29%,表现为在SM琼脂上形成的清除带直径减小。此外,在处理D中添加芽孢杆菌可显著提高副溶血性弧菌攻毒后蒿的存活率(P<0.05)。这表明芽孢杆菌有可能作为一种候选益生菌来预防副溶血性弧菌引起的疾病。
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6 weeks
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