一株溶藻弧菌H1B6的分离与鉴定
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摘要
从患病的半滑舌鳎(Cynoglossus semilaevis)溃疡体表处筛选到一株H1B6优势菌株,根据形态观察、16S rDNA检测、生理生化鉴定结果,H1B6菌株与溶藻弧菌最为接近,而在以16S rDNA序列构建的系统发育进化树模型中,H1B6菌株也与溶藻弧菌菌株聚为一支。Biolog微生物鉴定结果显示,H1B6菌株为溶藻弧菌的可能性为98.6%,确定H1B6菌株为溶藻弧菌。斑马鱼攻毒试验结果显示,H1B6菌株24 h的半数致死浓度(LD_(50))为4.58×10~6 CFU/mL。
A dominant strain of H1B6 was isolated from the ulcer surface of diseased Cynoglossus semilaevis. According to morphological observation, 16 S rDNA sequencing, physiological and biochemical identification results, H1B6 strain was suspected as Vibrio alginolyticus. According to the phylogenetic tree model constructed by 16 S rDNA sequence, H1B6 strain was clustered with Vibrio alginolyticus strains. Furthermore, H1B6 strain was finally identified as Vibrio alginolyticus by using Biolog microbial identification test,and the probability(PROB value) was 98.6%. H1B6 strain's acute toxicity test on zebrafish was performed and half lethal concentration(LD_(50)) after 24 hours was determined as 4.58×10~6 CFU/mL.
A dominant strain of H1B6 was isolated from the ulcer surface of diseased Cynoglossus semilaevis. According to morphological observation, 16 S rDNA sequencing, physiological and biochemical identification results, H1B6 strain was suspected as Vibrio alginolyticus. According to the phylogenetic tree model constructed by 16 S rDNA sequence, H1B6 strain was clustered with Vibrio alginolyticus strains. Furthermore, H1B6 strain was finally identified as Vibrio alginolyticus by using Biolog microbial identification test,and the probability(PROB value) was 98.6%. H1B6 strain's acute toxicity test on zebrafish was performed and half lethal concentration(LD_(50)) after 24 hours was determined as 4.58×10~6 CFU/mL.
引文
[1] 周红霞.半滑舌鳎溃疡相关气单胞菌的鉴定及其致病性研究[D].贵阳:贵州大学,2017.
[2] 徐晓丽,邵蓬,李贺密,等.半滑舌鳎体表溃疡病病原的分离鉴定[J].中国海洋大学学报(自然科学版),2015,45(11):29-35.
[3] 高桂生,张艳英,吉志新,等.半滑舌鳎致病性荧光假单胞菌的分离鉴定及其感染的病理损伤[J].中国兽医学报,2016,36(7):1145-1150.
[4] 王燕.两株海洋病原菌的分离鉴定、致病机理研究及三株海洋新菌的分类鉴定[D].青岛:中国海洋大学,2010.
[5] HOLT J G,KRIEG N R,SNEATH P H A,et al.Bergey’s manual ofdeterminative bacteriology [M].9th ed.Maryland:Williams& Wilkins,1994.
[6] 李翠萍,吴民耀,王宏元.3种半数致死浓度计算方法之比较[J].动物医学进展,2012,33(9):89-92.
[7] 聂忠学,张谷若,聂晶,等.溶藻弧菌的生化特征与中毒表现[J].贵州医药,1995(4):252-253.
[8] 祝玲,吴忠业,林阿乞,等.引发杂色鲍苗掉板死亡的溶藻弧菌分离鉴定及毒力分析[J].水利渔业,2008,28(3):97-99.
[9] 丁云娟,彭勇,林洪,等.一株副溶血弧菌qdfsVp001的分离与鉴定[J].海洋湖沼通报,2012(3):44-49.
[10] 苗鹏飞,杨映,谭淑雯,等.罗氏沼虾致病性溶藻弧菌的鉴定及药敏分析[J].水产科学,2018,37(3):384-388.
[11] 付汉清,林茂,翟少伟,等.一株宽谱裂解性溶藻弧菌噬菌体ФV170的分离鉴定及生物学特性[J/OL].微生物学通报,2018-06-25[2018-12-16].https://kns.cnki.net/kcms/detail/11.1996.Q.20180622.1513.004.html.DOI:10.13344/j.microbiol.china.180244.
[12] 焦彦凯,严小军,李小兵.溶藻细菌及溶藻化合物研究进展[J].工业微生物,2018,48(4):56-62.
[13] LV T T,SONG T X,LIU H J,et al.Isolation and characterization of a virulence related Vibrio alginolyticus strain Wz11 pathogenic to cuttlefish,Sepia pharaonis[J].Microbial pathogenesis,2019,126:165-171.
[14] 傅超英,王建平,孙琛,等.大黄鱼主要致病菌拮抗菌株的分离鉴定、抑菌谱及安全性分析[J/OL].生物技术通报,2018-09-25[2018-12-16].http://kns.cnki.net/kcms/detail/11.2396.Q.20180921.1144.004.html.DOI:10.13560/j.cnki.biotech.bull.1985.2018-0565.
[15] 姜燕,崔晓翠,张正,等.养殖大菱鲆肠炎病的流行病学和病原学研究[J].安徽农业科学,2016,44(17):123-126.
[16] XIE Z Y,KE S W,HU C Q,et al.First characterization of bacterial pathogen,Vibrio alginolyticus,for Porites andrewsi White syndrome in the South China Sea[J].PLoS One,2013,8(9):1-8.
[17] 李亚军,耿毅,余泽辉,等.两株不同血清型鱼源无乳链球菌生物学特性的比较研究[J].中国预防兽医学报,2018,40(12):1122-1127.
[18] 高艳侠,张德锋,可小丽,等.罗非鱼源无乳链球菌肠道拮抗芽孢杆菌的筛选及其生物学特性[J/OL].微生物学报:2018-11-29[2019-01-05].https://kns.cnki.net/kcms/detail/11.1995.Q.20181127.1603.014.html.DOI:10.13343/j.cnki.wsxb.20180368.
[2] 徐晓丽,邵蓬,李贺密,等.半滑舌鳎体表溃疡病病原的分离鉴定[J].中国海洋大学学报(自然科学版),2015,45(11):29-35.
[3] 高桂生,张艳英,吉志新,等.半滑舌鳎致病性荧光假单胞菌的分离鉴定及其感染的病理损伤[J].中国兽医学报,2016,36(7):1145-1150.
[4] 王燕.两株海洋病原菌的分离鉴定、致病机理研究及三株海洋新菌的分类鉴定[D].青岛:中国海洋大学,2010.
[5] HOLT J G,KRIEG N R,SNEATH P H A,et al.Bergey’s manual ofdeterminative bacteriology [M].9th ed.Maryland:Williams& Wilkins,1994.
[6] 李翠萍,吴民耀,王宏元.3种半数致死浓度计算方法之比较[J].动物医学进展,2012,33(9):89-92.
[7] 聂忠学,张谷若,聂晶,等.溶藻弧菌的生化特征与中毒表现[J].贵州医药,1995(4):252-253.
[8] 祝玲,吴忠业,林阿乞,等.引发杂色鲍苗掉板死亡的溶藻弧菌分离鉴定及毒力分析[J].水利渔业,2008,28(3):97-99.
[9] 丁云娟,彭勇,林洪,等.一株副溶血弧菌qdfsVp001的分离与鉴定[J].海洋湖沼通报,2012(3):44-49.
[10] 苗鹏飞,杨映,谭淑雯,等.罗氏沼虾致病性溶藻弧菌的鉴定及药敏分析[J].水产科学,2018,37(3):384-388.
[11] 付汉清,林茂,翟少伟,等.一株宽谱裂解性溶藻弧菌噬菌体ФV170的分离鉴定及生物学特性[J/OL].微生物学通报,2018-06-25[2018-12-16].https://kns.cnki.net/kcms/detail/11.1996.Q.20180622.1513.004.html.DOI:10.13344/j.microbiol.china.180244.
[12] 焦彦凯,严小军,李小兵.溶藻细菌及溶藻化合物研究进展[J].工业微生物,2018,48(4):56-62.
[13] LV T T,SONG T X,LIU H J,et al.Isolation and characterization of a virulence related Vibrio alginolyticus strain Wz11 pathogenic to cuttlefish,Sepia pharaonis[J].Microbial pathogenesis,2019,126:165-171.
[14] 傅超英,王建平,孙琛,等.大黄鱼主要致病菌拮抗菌株的分离鉴定、抑菌谱及安全性分析[J/OL].生物技术通报,2018-09-25[2018-12-16].http://kns.cnki.net/kcms/detail/11.2396.Q.20180921.1144.004.html.DOI:10.13560/j.cnki.biotech.bull.1985.2018-0565.
[15] 姜燕,崔晓翠,张正,等.养殖大菱鲆肠炎病的流行病学和病原学研究[J].安徽农业科学,2016,44(17):123-126.
[16] XIE Z Y,KE S W,HU C Q,et al.First characterization of bacterial pathogen,Vibrio alginolyticus,for Porites andrewsi White syndrome in the South China Sea[J].PLoS One,2013,8(9):1-8.
[17] 李亚军,耿毅,余泽辉,等.两株不同血清型鱼源无乳链球菌生物学特性的比较研究[J].中国预防兽医学报,2018,40(12):1122-1127.
[18] 高艳侠,张德锋,可小丽,等.罗非鱼源无乳链球菌肠道拮抗芽孢杆菌的筛选及其生物学特性[J/OL].微生物学报:2018-11-29[2019-01-05].https://kns.cnki.net/kcms/detail/11.1995.Q.20181127.1603.014.html.DOI:10.13343/j.cnki.wsxb.20180368.