耐盐好氧反硝化菌黏质沙雷氏菌HL4的分离鉴定及脱氮性能  

Characterization and Nitrogen Removal Performance of a Salt-tolerant Aerobic Denitrifying Bacterium Serratia marcescens HL4

作　　者：刘凤坤 赵吉臣 许敬轩 郑雅之 鲁慧杰 孙育平 阮灼豪 郭慧[1] 杨俊贤 黄文 LIU Fengkun;ZHAO Jichen;XU Jingxuan;ZHENG Yazhi;LU Huijie;SUN Yuping;RUAN Zhuohao;GUO Hui;YANG Junxian;HUANG Wen(College of fisheries,Guangdong Ocean University,Zhanjiang 524088,China;Institute of Animal Science,Guangdong Academy of Agricultural Sciences/Guangdong Key Laboratory of Animal Breeding and Nutrition/South China Key Laboratory of Animal Nutrition and Feed,Ministry of Agriculture and Rural Areas/Aquatic Research Center Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;Pingsha Town Social Affairs Service Center,Jinwan District,Zhuhai 519055,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088 [2]广东省农业科学院动物科学研究所/广东省畜禽育种与营养研究重点实验室/农业农村部华南动物营养与饲料重点实验室/广东省农业科学院水产研究中心,广东广州510640 [3]珠海市金湾区平沙镇社会事务服务中心,广东珠海519055

出　　处：《广东海洋大学学报》2024年第4期38-46,共9页Journal of Guangdong Ocean University

基　　金：广东省农业科学院人才项目(R2020YJ-QG001,R2022PY-QY008);广东省农业科学院协同创新中心课题(202137,202138,XT202231);珠海市斗门区白蕉镇政府服务项目(ZHWZ2021-403FW,ZHWZ2021-408FW);乡村振兴战略专项资金(农业科技能力提升)项目(2023工作站12)。

摘　　要：【目的】分离鉴定一株耐盐好氧反硝化细菌,以开发高效脱氮菌种,加强海水及咸淡水养殖尾水脱氮技术研究。【方法】采集花鲈(Lateolabrax maculatus)池塘原位底泥,采用稀释涂布法初步分离一株脱氮功能最佳的菌株HL4,通过16s rDNA测序以及生理生化实验鉴定菌株,通过设置不同碳源、碳氮质量比(m_(C)/m_(N))、pH、温度、盐度等条件探明其最适培养条件及脱氮性能。通过攻毒试验[以健康斑马鱼(Danio rerio)为受试生物],检验菌株的环境安全性。【结果】经鉴定,菌株HL4为黏质沙雷氏菌(Serratia marcescens),命名为S.marcescens HL4。以葡萄糖或琥珀酸钠为碳源,菌株HL4在m_(C)/m_(N)=20、pH=6.0~7.0、温度30℃的条件下具有较强脱氮能力,48 h TN去除率为81.83%~87.17%;该菌株在0~30的盐度环境下均可生长,在0~10盐度范围内72 h的总氮(TN)去除率为68.76%~76.59%,在20~30盐度范围内72 h的TN去除率为23.84%~53.94%;在NH_(4)^(+)-N质量浓度为0~100 mg·L^(-1)时,72 h NH_(4)^(+)-N去除率皆在90%以上,在NH_(4)^(+)-N质量浓度为500 mg·L^(-1)时,72 h NH_(4)^(+)-N去除率高达73.52%。健康斑马鱼在含1×10^(6)CFU·mL^(-1)菌株HL4的水体中15 d后,成活率100%。【结论】菌株HL4生物及环境安全性较高,在处理中高浓度铵废水以及治理咸淡水、低盐度海水养殖尾水等领域具有潜在的应用价值。【Objective】 To isolate and identify an efficient salt tolerant aerobic denitrifying bacterial strain and strengthen the research of denitrization technology in seawater and brackish water culture.【Method】 Using the in-situ sediment of Lateolabrax maculatus pond as screening objects,a strain HL4 with the best denitrification function was initially isolated by dilution coating method.The strain was identified by 16s rDNA sequencing and physiological and biochemical experiments.The optimal culture conditions and denitrifying performance were investigated by setting different carbon sources,carbon nitrogen mass ratio(m_C/m_N),pH,temperature,salinity and other conditions.The environmental safety of the strain was tested by challenge test(with healthy zebrafish,Danio rerio,as the test organism).【Result】 The strain HL4 was identified as Serratia marcescens,named S.marcescens HL4.Using glucose or sodium succinate as carbon source,the strain HL4 had strong nitrogen removal ability at m_C/m_(N) = 20,pH = 6.0-7.0,and temperature 30 ℃,48 h TN removal rate was 81.83%-87.17%.The strain could grow in 0-30 salinity environment and had a certain nitrogen removal ability.The 72 h TN removal rate at 0-10 salinity was 68.76%-76.59%,and the 72 h TN removal rate at 20-30 salinity was 23.84%-53.94%.When the concentration of NH_(4)^(+)-N was 0-100 mg·L^(-1),the 72 h removal rate of NH_(4)^(+)-N was above 90%;when the concentration of NH_(4)^(+)-N was 500 mg·L^(-1),the 72 h removal rate of NH_(4)^(+)-N is as high as 73.52%.After 15 days in water containing 1 × 10^(6) CFU·mL^(-1 )strain HL4,the survival rate of healthy zebrafish was 100%.【Conclusion】 Strain HL4 has high biosafety and environmental safety,and has potential application value in the treatment of high-concentration ammonium wastewater,brackish water and low-salinity aquaculture tail water.

关 键 词：黏质沙雷氏菌 好氧反硝化 耐盐 生物安全性 环境因子 脱氮能力 

分 类 号：S917.1[农业科学—水产科学]