从贝类中分离出的本地细菌作为抗致病性弧菌潜在益生菌的筛选与评估。
Screening and evaluation of local bacteria isolated from shellfish as potential probiotics against pathogenic Vibrios.
作者信息
Jasmin M Y, Wagaman Hazimah, Yin Tan Ai, Ina-salwany M Y, Daud H M, Karim Murni
出版信息
J Environ Biol. 2016 Jul;37(4 Spec No):801-9.
The present study was carried out to isolate, screen and evaluate potential candidates of local bacteria isolated from tiger shrimp Penaeus monodon and slipper cupped oysters Crassostrea iredalei as probiotics in shellfish aquaculture. A total of 144 of bacteria were successfully isolated from the intestine and stomach of 20 tails of healthy adult tiger shrimp P. monodon, while 136 were successfully isolated from the digestive tract, gills and inner shells of 10 healthy adult C. iredalei. The number of potential isolates was narrowed down to two from tiger shrimp, and one from slipper cupped oyster after in vitro screening assays. The three isolates, labeled as G11, I24 and S66, were identified as Virgibacillus sp., Bacillus sp. and Exiquobacterium sp., respectively, using 16S rDNA gene analysis. The antagonistic ability of the isolates towards Vibrio alginolyticus and Vibrio harveyi were conducted in stagnant and liquid modes via spot lawn and broth co-culture assay, respectively. In these assays, all the potential probionts were inhibitory to both pathogenic vibrios. In the in-vivo assay, Artemia was used as host and treated with different concentrations of potential probionts (10(4), 10(6) and 10(8) CFU ml(-1)), and challenged with V. alginolyticus and V. harveyi at 105 CFU ml(-1), respectively. Artemia treated with probiont G11 at all concentrations and challenged with V. alginolyticus had increased survival (70 ? 80 %), which was significantly higher as compared with group with only the pathogen (20 %). Meanwhile, probiont I24 increased the survival of Artemia by 70 % at a concentration of 10(8) CFU ml(-1) after being challenged with V. alginolyticus and Artemia treated with 10(6) CFU ml(-1) of probiont S66 had increased survival of 90% after being challenged with V. harveyi. Thus, the three isolates might have potential applications as probiotics in shellfish aquaculture against vibriosis. ?
本研究旨在分离、筛选和评估从斑节对虾（Penaeus monodon）和翡翠贻贝（Crassostrea iredalei）中分离出的本地细菌作为贝类水产养殖益生菌的潜在候选菌株。从20尾健康成年斑节对虾的肠道和胃中成功分离出144株细菌，同时从10只健康成年翡翠贻贝的消化道、鳃和内壳中成功分离出136株细菌。经过体外筛选试验后，斑节对虾的潜在分离株数量减少到两株，翡翠贻贝的减少到一株。使用16S rDNA基因分析，将这三株分离株分别鉴定为维氏芽孢杆菌（Virgibacillus sp.）、芽孢杆菌（Bacillus sp.）和短小芽孢杆菌（Exiquobacterium sp.），分别标记为G11、I24和S66。通过点种法和平板共培养试验分别在静置和液体模式下检测分离株对溶藻弧菌（Vibrio alginolyticus）和哈维氏弧菌（Vibrio harveyi）的拮抗能力。在这些试验中，所有潜在益生菌对两种致病性弧菌均有抑制作用。在体内试验中，卤虫被用作宿主，用不同浓度的潜在益生菌（10⁴、10⁶和10⁸ CFU ml⁻¹）处理，然后分别用10⁵ CFU ml⁻¹的溶藻弧菌和哈维氏弧菌进行攻毒。用益生菌G11各浓度处理并受到溶藻弧菌攻毒的卤虫存活率提高（70% - 80%），与仅感染病原体的组（20%）相比显著更高。同时，在用10⁸ CFU ml⁻¹浓度的益生菌I24处理后受到溶藻弧菌攻毒的卤虫存活率提高了70%，在用10⁶ CFU ml⁻¹的益生菌S66处理后受到哈维氏弧菌攻毒的卤虫存活率提高了90%。因此，这三株分离株作为益生菌在贝类水产养殖中对抗弧菌病可能具有潜在应用价值。
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