Biological Characteristics and Application in Water Purification in Sea Cucumber Culture Ponds in Cryotolerant Degrading Bacterium Strain Bacillus mobilis 10
TANG Wei1,2,3, WANG Qi3, ZHANG Jun2,3, LI Jiaxin2, XING Wei3, HE Zengguo1,3
1. Marine Microbiology Engineering Research and Development Center, Marine Biomedical Research Institute of Qingdao, Qingdao 266100, China; 2. School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China; 3. Qingdao Bioantai Biotechnology Limited Company, Qingdao 266100, China
Abstract：A bacterium strain collected from the coastal soil of Qingdao was isolated and purified after 48 h of culture at 15 °C with 7 media, and was identified as Bacillus mobilis by morphological observation, physicochemical characteristics and 16S rRNA gene sequence analysis. The strain was inoculated into LB liquid culture at 5, 28 and 37 °C and 0, 2%, 4%, 6% and 8% of NaCl to explore the temperature and salinity tolerance of the B. mobilis, and the strain was sprayed once every 3 days in a sea cucumber Apostichopus japanicus seedling pond with a final concentration of about 1.0×104 cfu/mL to purify the water quality in the sea cucumber culture pond, so as to realize the industrialization of this strain.The transparent circle method showed that B. mobilis 10 produced protease, amylase and cellulase activities at low temperature conditions (15 °C), with protease activity of 9.05 U/mL after 56 h of fermentation and NaCl tolerance of up to 8%.The test of degradation ability of the strain to dissolved substances showed that the degradation rate in the feed culture medium was observed to be (36.72±0.53)% in protein and (22.80±0.81)% in (chemical oxygen demand) COD using sea cucumber feed culture medium, 8 h after inoculation. Pilot-scale (50 L) fermentation techniques was established to microbial preparation, and applied in the sea cucumber breeding. Dynamic monitoring of the bacterial activity of B. mobilis 10 in sea cucumber culture ponds for 8 days revealed that both the contents of protein and COD in sea cucumber ponds were significantly reduced by sprinkling the bacterial solution with the final concentration of 1.0×104 cfu/mL. The findings had exemplified the successful exploration case regarding the development of microbial products for use in aquaculture practice.
[1] 农业农村部渔业渔政管理局,全国水产技术推广总站,中国水产学会编制.2021中国渔业统计年鉴[M].北京:中国农业出版社,2021:21-23. [2] 中华人民共和国农业部.刺参配合饲料:SC/T 2037—2006[S].北京:中国农业出版社,2006. [3] 闫法军,田相利,董双林,等.一株低温降解菌的分离鉴定及其降解特性研究[J].中国海洋大学学报(自然科学版),2011,41(3):24-30. [4] 赵斌,李成林,周红学,等.海参产业绿色发展对策研究[J].中国海洋经济,2021(1):22-42. [5] 廖梅杰,王印庚,李彬,等.我国海参养殖产业现状、存在问题及对策探讨(下)[J].科学养鱼,2021(4):26-27. [6] 杜金艳.一株低温芽孢杆菌净水剂的应用研究[D].大连:大连工业大学,2017. [7] 窦赫喆.海参养殖环境中水体净化微生物的筛选及鉴定[D].大连:大连工业大学,2014. [8] 姜松,王军红,范嗣刚,等.添加光合细菌对糙海参幼苗培育阶段的影响研究[J].南方水产科学,2014,10(6):78-82. [9] 巩玉辉,陈翠翠,马玉堃,等.微生态制剂在海参养殖中的应用[J].中国饲料,2012(8):37-39. [10] YU H B, ZHANG C, GAO Q F, et al. Impact of water temperature on the growth and fatty acid profiles of juvenile sea cucumber Apostichopus japonicus (Selenka)[J]. Journal of Thermal Biology,2016,60:155-161. [11] LIU Y, DU J, LAI Q L, et al. Proposal of nine novel species of the Bacillus cereus group[J]. International Journal of Systematic and Evolutionary Microbiology,2017,67(8):2499-2508. [12] 史晓玲.去除水体中重金属芽孢杆菌属细菌的筛选及机制初探[D].昆明:云南大学,2021. [13] 王琪鑫.生物发酵法制备甲壳素的研究[D].青岛:青岛科技大学,2021. [14] 江世杰,韦廷舟,文怡,等.一株运动芽孢杆菌Bacillus mobilis在增强油菜耐盐促生中的应用:CN114591855B[P].2023-05-26. [15] 谢承卫,王应兰,李小军,等.一种运动芽孢杆菌及其应用:CN112760270B[P].2022-06-14. [16] 闫法军,田相利,董双林,等.刺参养殖池塘降解有机污染物常、低温芽孢杆菌的分离筛选[J].中国海洋大学学报(自然科学版),2013,43(6):17-24. [17] 国家药典委员会.中华人民共和国药典-三部:2015年版[M].北京:中国医药科技出版社,2015:3524-3525. [18] 雷衍之.养殖水环境化学实验[M].北京:中国农业出版社,2006:31-86. [19] 陈峰.耐低温脱氮除磷菌的筛选鉴定及复合菌剂的构建研究[D].沈阳:沈阳大学,2022. [20] 董怡华,张雪莹,邹立安,等.耐低温好氧反硝化菌Aeromonas sp.的分离鉴定及脱氮条件优化[J].微生物学报,2022,62(6):2038-2052. [21] 韦廷舟,文怡,王超,等.一株产IAA芽孢杆菌ST37对油菜的耐盐促生作用[J].江苏农业科学,2023,51(6):210-215. [22] 胡玉婕,朱秀玲,丁延芹,等.芽孢杆菌的耐盐促生机制研究进展[J].生物技术通报,2020,36(9):64-74. [23] MA Q S, HE Z G. Screening and characterization of nitrite-degrading bacterial isolates using a novel culture medium[J]. Journal of Ocean University of China,2020,19(1):241-248. [24] 胡婷,马青山,贠锦军,等.一株枯草芽孢杆菌新菌株,微生态制剂及应用:CN105441348B[P].2019-05-21. [25] 刘晓燕,王玲玲,栾会妮,等.一株枯草芽孢杆菌的分离鉴定、生物学特性及其对水质净化的作用[J].微生物学通报,2021,48(2):449-461. [26] 管立平,谢晶晶,李伟豪,等.一种赖氨酸芽孢杆菌对凡纳滨对虾生长和水质影响[J].渔业现代化,2022,49(6):77-83. [27] THURLOW C M, WILLIAMS M A, CARRIAS A, et al. Bacillus velezensis AP193 exerts probiotic effects in channel catfish (Ictalurus punctatus) and reduces aquaculture pond eutrophication[J]. Aquaculture,2019,503:347-356. [28] 罗毅志,施伟达,周冬仁,等.地衣芽孢杆菌分离株DSY002-2011对养殖水体氨氮与亚硝酸盐降解特性研究[J].现代农业科技,2014(3):236. [29] 王国旭,吴又多,胡馨月,等.水产养殖饲用微生态制剂研究进展[J].微生物学杂志,2022,42(6):118-128.
2024, Vol. 43 
