虾蟹池塘有机物降解菌富集驯化与分离筛选的初步研究  

A Primary Study on Enrichment, Domestication, Isolation and Screening of Organic-pollutant-degrading Bacteria in Ponds of Shrimp and Crab

作　　者：李步先 田相利[2] 董双林[2] 汪多 LI Buxian;TIAN Xiangli;DONG Shuanglin;WANG Duo(Tangshan Polytechnic College, Tangshan 063299;The Key Laboratory of Mariculture of Ministry of Education,Ocean University of China, Qingdao 266003;Suzhou Vocational and Technical College, Suzhou 234000)

机构地区：[1]唐山工业职业技术学院,河北唐山063299 [2]中国海洋大学海水养殖教育部重点实验室,山东青岛266003 [3]宿州职业技术学院,安徽宿州234000

出　　处：《海洋湖沼通报》2019年第3期143-153,共11页Transactions of Oceanology and Limnology

基　　金：国家“十二五”科技支撑计划项目(2011BAD13B03);山东省杰出青年基金项目(JQ201009)资助

摘　　要：采集虾蟹混养池塘沉积物,利用特定饵料培养基,并通过定期增加培养基中饵料添加量的方式,对沉积物中的复合菌进行了6个周期(7d/周期)的富集驯化培养。在COD含量约为1 000mg/L、菌接种量为10%的饵料降解溶液中,后期复合菌在5d内对COD的最大去除率分别达到了64%、70%和88%。在6个周期富集驯化结束后,从复合菌中分离出16株菌,经16S rRNA分析,其中8株属于弧菌属(vibrio sp.),1株属于芽孢杆菌目(Bacillales sp.),2株属于芽孢杆菌属(Bacillus sp.),2株属于交替假单胞菌属(Pseudoalteromonas sp.),1株属于气单胞菌属(Aeromonas sp.),1株属于希瓦氏菌属(Shewanella sp.)。经血平板初步筛选出4株不具潜在致病性的菌,1株属于芽孢杆菌目,1株为巨大芽孢杆菌(Bacillus megaterium),2株为交替假单胞菌,在菌接种量为1.0×10^7cfu/mL的降解溶液中,各株菌在5d内对COD的最大去除率分别达到了40%、40%、56%和46%。将这4株菌按一定比例组合后,在5d内对COD的最大去除率可达到72%。研究表明,经富集驯化后的复合菌及从中分离筛选出的4株不具潜在致病性的细菌及按一定比例重组后的菌在调控养殖环境有机物污染方面都具有潜在应用价值。The complex bacteria collected from the sediment of culture ponds for shrimp and crab were enriched and domesticated for six cycles, 7 days each cycle, by adding the shrimp feed to specific culture medium regularly. When the fourth, fifth or sixth cycle was finished, the content of organic pollutants(COD) in the feed was about 1000 mg/L and the inoculation size was controlled at 10%, the highest COD removal rates of 64%, 70%, or 88% within 5 days of the cultivation were recorded, respectively. At the end of all the cycles, 16 bacterial strains were isolated from the complex bacteria. According to 16 S rRNA gene sequence similarity, 8 of them were identified as Vibrio sp., 1 as Bacillales, 2 as Bacillus sp., 2 as Pseudoalteromonas sp., 1 as Aeromonas sp. and 1 as Shewanella sp.. After preliminarily screening using blood plates, 4 bacterial strains that were not potential pathogens were selected, one of them was Bacillales, one of them was Bacillus sp. and two of them were Pseudoalteromonas sp.. In the feed with an inoculation of 1.0×10^7cfu/mL, the 4 bacteria were able to degrade the organic compounds in the feed and the highest COD removal rates of 40%, 40%, 56% and 46% within 5 days were recorded, respectively. When the 4 bacterial strains were combined in suitable proportion, the highest COD removal rate of 72% within 5 days was recorded. Therefore, it can be concluded that both of the complex bacteria obtained after enrichment and domestication and the 4 bacteria obtained after isolation and screening have potential application in pollution control for culture ponds of shrimp and crab.

关 键 词：虾蟹池塘 有机物降解菌 富集驯化 分离筛选 

分 类 号：Q815[生物学—生物工程]