一株发光细菌的分离鉴定及其对有毒物质的毒性响应
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摘要
从新鲜海鱼肠道内分离获得1株发光细菌Fc-11-1,通过菌落形态特征、生理生化特征、16SrDNA序列分析及系统发育树分析对其进行鉴定,初步确定发光细菌Fc-11-1为Vibrio toranzoniae。从发光细菌Fc-11-1的生长和发光强度两方面,初步探索了发光细菌Fc-11-1对食品中5种常见有毒物质百菌清、呋喃唑酮、红霉素、氯霉素、阿维菌素的毒性响应。结果表明,发光细菌Fc-11-1对5种有毒物质均有不同程度的毒性响应,可用于食品中有毒物质残留的检测。
We isolated a luminous bacterium Fc-11-1 from the intestinal tract of fresh marine fish.Moreover,we identified that the luminous bacterium Fc-11-1 belongs to Vibrio toranzoniae by colony morphology,physiological and biochemical characteristics,16 SrDNA sequencing analysis,and phylogenetic tree analysis.Furthermore,from the growth and luminescence intensity of the luminous bacterium Fc-11-1,we investigated the toxicity response of the luminous bacterium Fc-11-1 to five common hazardous materials,such as Chlorothaloni,Furazolidone,Chloramphenicol,Erythromycin,and Avermectin in food.The results show that the luminous bacterium Fc-11-1 has different degrees of toxicity response to five hazardous materials,which can be used for the detection of hazardous materials residues in food.
We isolated a luminous bacterium Fc-11-1 from the intestinal tract of fresh marine fish.Moreover,we identified that the luminous bacterium Fc-11-1 belongs to Vibrio toranzoniae by colony morphology,physiological and biochemical characteristics,16 SrDNA sequencing analysis,and phylogenetic tree analysis.Furthermore,from the growth and luminescence intensity of the luminous bacterium Fc-11-1,we investigated the toxicity response of the luminous bacterium Fc-11-1 to five common hazardous materials,such as Chlorothaloni,Furazolidone,Chloramphenicol,Erythromycin,and Avermectin in food.The results show that the luminous bacterium Fc-11-1 has different degrees of toxicity response to five hazardous materials,which can be used for the detection of hazardous materials residues in food.
引文
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[14]GB 2763-2014.食品安全国家标准食品中农药最大残留限量[S].
[15]罗文婷,吴青,简伟明,等.超高效液相色谱法测定水产品中违禁兽药氯霉素、呋喃唑酮和甲硝唑残留[J].分析化学,2009,37(6):877-880.LUO W T,WU Q,JIAN W M,et al.Stimultaneous determination of illegal veterinary drugs of Chloramphenicol,furazolidone and metronidazole in aquatic product using ultra performance liquid chromatography[J].Chinese Journal of Analytical Chemistry,2009,37(6):877-880.
[16]鲁顺保,张艳杰,陈成榕,等.基于BIOLOG指纹解析三种不同森林类型土壤细菌群落功能差异[J].土壤学报,2013,50(3):618-623.LU S B,ZHANG Y J,CHEN C R,et al.Analysis of functional differences between soil bacterial communities in three different types of forest soils based on BIOLOG fingerprint[J].Acta Pedologica Sinica,2013,50(3):618-623.
[17]廖先燕,张涛,曾军,等.一种基于BIOLOG代谢指纹分析的油气微生物勘探新方法[J].微生物学报,2012,52(4):505-511.LIAO X Y,ZHANG T,ZENG J,et al.A new method of microbial prospecting for oil and gas based on BIOLOG metabolic fingerprinting analysis[J].Acta Microbiologica Sinica,2012,52(4):505-511.
[18]张惠燕,李艳菊,顾金刚,等.基于Biolog-FF技术的金霉素降解真菌的碳代谢特征研究[J].微生物学通报,2015,42(7):1241-1247.ZHANG H Y,LI Y J,GU J G,et al.On carbon metabolism of fungi in Chlortetracycline degradation based on Biolog-FF system[J].Microbiology China,2015,42(7):1241-1247.
[19]张朝正,郭兰珍.利用16SrDNA序列分析和Biolog快速鉴定方法鉴定产脂肪酶菌株[J].河北工业大学学报,2009,38(5):52-55.ZHANG Z Z,GUO L Z.Identification of microbe with lipase activity by 16SrDNA sequence analysis and Biolog microstation system[J].Journal of Hebei University of Technology,2009,38(5):52-55.
[20]LASA A,DIEGUEZ A L,ROMALDE J L.Vibrio toranzoniae sp.nov.,a new member of the Splendidus clade in the genus Vibrio[J].Systematic and Applied Microbiology,2013,36(2):96-100.
[21]凌云,赵渝,徐亚同,等.发光细菌法在食品安全性检测中的应用[J].食品与生物技术,2005,24(6):106-110.LING Y,ZHAO Y,XU Y T,et al.Application of photobacteria in food safety inspection[J].Journal of Food Science and Biotechnology,2005,24(6):106-110.
[2]段效辉,王颖,曹鹏,等.发光细菌在水产品安全中的应用研究进展[J].化学与生物工程,2016,33(10):8-11.DUAN X H,WANG Y,CAO P,et al.Research progress of luminescent bacteria in safty of aquatic products[J].Chemistry&Bioengineering,2016,33(10):8-11.
[3]王亚群,王静雪,林洪,等.发光细菌法检测水产品中氯霉素体系的建立[J].中国海洋大学学报(自然科学版),2009,39(1):66-70.WANG Y Q,WANG J X,LIN H,et al.Construction of a system to detect Chloramphenicol residues in aquatic foods by luminescent bacteria[J].Periodical of Ocean University of China(Natural Science Edition),2009,39(1):66-70.
[4]吴淑杭,周德平,徐亚同,等.重金属汞、镉和铬对明亮发光杆菌的生物毒性[J].农业环境科学学报,2011,30(12):2446-2450.WU S H,ZHOU D P,XU Y T,et al.Toxicities of heavy metals Hg,Cd and Cr to Photobacterium phosphoreum[J].Journal of Agro-Environment Science,2011,30(12):2446-2450.
[5]汤云.发光细菌在重金属检测中的应用[J].仪器仪表与分析监测,2010(4):1-3.TANG Y.Luminous bacteria in heavy metal detection[J].Instrumentation Analysis Monitoring,2010(4):1-3.
[6]石颖,丁武,张志超,等.应用青海弧菌评价常见8种兽药的急性毒性[J].西北农业学报,2012,21(6):17-21.SHI Y,DING W,ZHANG Z C,et al.Determination of the acute toxicity of eight kinds of veterinary drug on V.qinghaiensis[J].Acta Agriculturae Boreali-Occidentalis Sinica,2012,21(6):17-21.
[7]束琴霞,张鹏,张林,等.发光细菌法快速检测水发产品中甲醛毒性的研究[J].江苏农业科学,2013,41(11):329-330,331.SHU Q X,ZHANG P,ZHANG L,et al.Study on rapid detection of formaldehyde emission toxicity in water soaked products by luminous bacteria[J].Jiangsu Agricultural Sciences,2013,41(11):329-330,331.
[8]许道艳,李伟,张芳,等.用发光细菌法监测海洋沉积物综合毒性的可行性研究[J].海洋环境科学,2009,28(5):570-572.XU D Y,LI W,ZHANG F,et al.Feasibility study on monitoring toxicity in marine sediment by luminescent bacteria test[J].Marine Environmental Science,2009,28(5):570-572.
[9]刘超,姚站馨.发光细菌用于环境检测的研究进展[J].军事医学科学院院刊,2009,33(2):179-182.LIU C,YAO Z X.Bioluminescent bacteria in environmental pollutants monitoring:research progress[J].Bulletin of the Academy of Military Medical Sciences,2009,33(2):179-182.
[10]HOLT J G,KRIEG N R,SNEATH P H A,et al.Bergey′s Manual of Determinative Bacteriology[M].9th ed.Baltimore:Williams&Wilkins,1994:71-377.
[11]杜宗军,王祥红,李海峰,等.一株海洋发光细菌的分离鉴定及其发光条件的初步研究[J].海洋湖沼通报,2003,33(2):58-63.DU Z J,WANG X H,LI H F,et al.Isolation and identification of a luminous bacterium D40and primary studies on its luminescent conditions[J].Transactions of Oceanology and Limnology,2003,33(2):58-63.
[12]周德庆.微生物学实验手册[M].上海:上海科学技术出版社,1986:27-30.
[13]范秀容,李广武,沈萍.微生物学实验[M].北京:海洋出版社,1999:166-190.
[14]GB 2763-2014.食品安全国家标准食品中农药最大残留限量[S].
[15]罗文婷,吴青,简伟明,等.超高效液相色谱法测定水产品中违禁兽药氯霉素、呋喃唑酮和甲硝唑残留[J].分析化学,2009,37(6):877-880.LUO W T,WU Q,JIAN W M,et al.Stimultaneous determination of illegal veterinary drugs of Chloramphenicol,furazolidone and metronidazole in aquatic product using ultra performance liquid chromatography[J].Chinese Journal of Analytical Chemistry,2009,37(6):877-880.
[16]鲁顺保,张艳杰,陈成榕,等.基于BIOLOG指纹解析三种不同森林类型土壤细菌群落功能差异[J].土壤学报,2013,50(3):618-623.LU S B,ZHANG Y J,CHEN C R,et al.Analysis of functional differences between soil bacterial communities in three different types of forest soils based on BIOLOG fingerprint[J].Acta Pedologica Sinica,2013,50(3):618-623.
[17]廖先燕,张涛,曾军,等.一种基于BIOLOG代谢指纹分析的油气微生物勘探新方法[J].微生物学报,2012,52(4):505-511.LIAO X Y,ZHANG T,ZENG J,et al.A new method of microbial prospecting for oil and gas based on BIOLOG metabolic fingerprinting analysis[J].Acta Microbiologica Sinica,2012,52(4):505-511.
[18]张惠燕,李艳菊,顾金刚,等.基于Biolog-FF技术的金霉素降解真菌的碳代谢特征研究[J].微生物学通报,2015,42(7):1241-1247.ZHANG H Y,LI Y J,GU J G,et al.On carbon metabolism of fungi in Chlortetracycline degradation based on Biolog-FF system[J].Microbiology China,2015,42(7):1241-1247.
[19]张朝正,郭兰珍.利用16SrDNA序列分析和Biolog快速鉴定方法鉴定产脂肪酶菌株[J].河北工业大学学报,2009,38(5):52-55.ZHANG Z Z,GUO L Z.Identification of microbe with lipase activity by 16SrDNA sequence analysis and Biolog microstation system[J].Journal of Hebei University of Technology,2009,38(5):52-55.
[20]LASA A,DIEGUEZ A L,ROMALDE J L.Vibrio toranzoniae sp.nov.,a new member of the Splendidus clade in the genus Vibrio[J].Systematic and Applied Microbiology,2013,36(2):96-100.
[21]凌云,赵渝,徐亚同,等.发光细菌法在食品安全性检测中的应用[J].食品与生物技术,2005,24(6):106-110.LING Y,ZHAO Y,XU Y T,et al.Application of photobacteria in food safety inspection[J].Journal of Food Science and Biotechnology,2005,24(6):106-110.