一株分离自丽江野鼠疫源地鼠疫噬菌体裂解特性的研究
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摘要
目的测定一株分离自丽江野鼠鼠疫疫源地鼠巢土的野生型鼠疫噬菌体(命名为LJ-7)的裂解谱,分析LJ-7的裂解能力及特点。方法采用双层平板法观察LJ-7对248株菌(包括10株云南鼠疫耶尔森菌、1株鼠疫疫苗株EV76、6株假结核耶尔森菌、37株致病小肠结肠炎耶尔森菌、17株非致病小肠结肠炎耶尔森菌、100株大肠埃希菌和77株其他菌株)在28℃、37℃及18℃3种温度时的裂解能力。结果 LJ-7对10株云南鼠疫耶尔森菌、1株鼠疫疫苗株EV76及1株弗氏志贺菌有裂解作用;对其余236株菌均不裂解。在37℃时LJ-7较鼠疫诊断噬菌体裂解性高,而在28℃时LJ-7较鼠疫诊断噬菌体裂解性低。结论 LJ-7在28℃、37℃及18℃时均能裂解鼠疫耶尔森菌,对弗氏志贺菌也有裂解性作用。表明LJ-7裂解细菌谱较宽,在不依赖鼠疫菌的情况下可由其他细菌提供营养而存在,它的发现对我国鼠疫疫源地环境微生态学和流行病学研究具有重要意义。
Objectives To determine the lytic spectrum of a wild type of plague phage(named LJ-7)isolated from the nesting soil of wild rats from plague foci in Lijiang and to analyze the lytic ability and characteristics of LJ-7. MethodsThe morphology,size,and titer of LJ-7 were determined using electron microscopy and the double-plate method.The double-plate method was used to observe the lytic ability of LJ-7 on 248 strains of bacteria(including 10 strains of Yersinia pestis,1 plague vaccine strain(Y.pseudotuberculosis),37 strains of pathogenic Y.enterocolitica,Y.pestis,17 strains of non-pathogenic Y.enteritidis,100 strains of Escherichia coli,and 77 other strains)at three temperatures:28℃,37 ℃,and 18 ℃. Results The plaque formed by LJ-7 had a distinct center and edges,the diameter of a single spot was 1-2 mm,and the titer was>1020 PFU/ml.The tail length was about 800 nm and the head diameter was 10-20 nm.LJ-7 presumably belongs to a long-tailed phage family(Styloviridae).LJ-7 displayed specific lytic ability on 10 Yunnan strains of Y.pestis,strain EV76,and 1 strain of Shigella flexneri,but it did not lyse the 236 other strains.LJ-7 was more cleavable at 37 ℃than a bacteriophage used to diagnosis plague but was less cleavable at 28 ℃. Conclusion The LJ-7 phage displayed lytic ability on Y.pestis at 28,37,and 18℃,and it also had a lytic effect on Shigella flexneri.This indicates that LJ-7 has a broad bacteriolytic spectrum and that LJ-7 can feed on other bacteria independent of Y.pestis.The discovery of this phage is of great significance to the environmental microecology and epidemiology of plague foci in China.
Objectives To determine the lytic spectrum of a wild type of plague phage(named LJ-7)isolated from the nesting soil of wild rats from plague foci in Lijiang and to analyze the lytic ability and characteristics of LJ-7. MethodsThe morphology,size,and titer of LJ-7 were determined using electron microscopy and the double-plate method.The double-plate method was used to observe the lytic ability of LJ-7 on 248 strains of bacteria(including 10 strains of Yersinia pestis,1 plague vaccine strain(Y.pseudotuberculosis),37 strains of pathogenic Y.enterocolitica,Y.pestis,17 strains of non-pathogenic Y.enteritidis,100 strains of Escherichia coli,and 77 other strains)at three temperatures:28℃,37 ℃,and 18 ℃. Results The plaque formed by LJ-7 had a distinct center and edges,the diameter of a single spot was 1-2 mm,and the titer was>1020 PFU/ml.The tail length was about 800 nm and the head diameter was 10-20 nm.LJ-7 presumably belongs to a long-tailed phage family(Styloviridae).LJ-7 displayed specific lytic ability on 10 Yunnan strains of Y.pestis,strain EV76,and 1 strain of Shigella flexneri,but it did not lyse the 236 other strains.LJ-7 was more cleavable at 37 ℃than a bacteriophage used to diagnosis plague but was less cleavable at 28 ℃. Conclusion The LJ-7 phage displayed lytic ability on Y.pestis at 28,37,and 18℃,and it also had a lytic effect on Shigella flexneri.This indicates that LJ-7 has a broad bacteriolytic spectrum and that LJ-7 can feed on other bacteria independent of Y.pestis.The discovery of this phage is of great significance to the environmental microecology and epidemiology of plague foci in China.
引文
[1]李玉贵,梁占凯,李建法,等.鼠疫防控实用技术病原学篇[M].北京:人民卫生出版社,2016:1.
[2]Hendrix RW.Bacteriophages:evolution of the majiority[J].Theor Popul Biol,2002(61):471-80.
[3]李存香,李伟,王鹏,等.1株温和噬菌体的发现及其流行病学意义[J].医学动物防制,2017,33(10):1073-5.
[4]李存香,王鹏,申小娜,等.野生型鼠疫噬菌体YPO60的分离和生物学特性鉴定[J].中华流行病学杂志,2016,37(6):873-6.
[5]金丽霞,于守鸿,祁芝珍,等.自四川石渠县分离3株野生型鼠疫噬菌体的鉴定[J].中国人兽共患病杂志,2001,17(5):11,124.
[6]宋延富.鼠疫菌以非典型形式在自然界长期保存的研究进展[J].中国地方病防治杂志,1995,10(2):101-3.
[7]张哲,李菁华,逯茵茵,等.粪肠球菌裂解性噬菌体φEn-ZZ8生物学特性的初步分析[J].微生物与感染,2015,10(4):241-6.
[8]张红明,黄庆,罗阳,等.肺炎克雷伯菌噬菌体的分离和生物学特性[J].中华医院感染学杂志,2008,18(5):608-11.
[9]邹玲,任慧英,杨超.一株宽裂解谱大肠埃希菌噬菌体的分离和裂解谱的测定及电镜观察[J].兽医科学,2014,3(5):118-120.
[10]贺雄,王虎.现代鼠疫概论[M].北京:科学出版社,2010:265.
[11]王梅,唐新元,粱莹,等.鼠疫菌基因组差异编码序列的筛选研究[J].中国病原生物学杂志,2017,12(2):115-8.
[12]李存香,靳娟,吴海莲,等.1978-2014年青海省果洛州鼠疫流行病学分析[J].中国病原生物学杂志,2017,12(7):651-3.
[13]潘光合,于建国.鼠疫杆菌在自然生态环境保存状况的研究进展[J].中国国境卫生检疫杂志,2010,33(3):214-6.
[14]何建,靳娟,张青雯,等.一株鼠毒素基因缺失突变鼠疫耶尔森氏菌株的构建和鉴定[J].中国病原生物学杂志,2016,11(6):513-6.
[15]俞东征.鼠疫与假结核耶尔森菌基因组比较分析[J].疾病监测,2011,26(10):760-5.
[16]宋志忠,夏连续,梁云,等.云南玉龙及古城区鼠疫自然疫源地判定及初步研究[J].中国地方病防治杂志,2008,23(1):3-7.
[17]苏丽琼,梁云,吴鹤松,等.云南省2014年鼠疫监测结果[J].中国地方病防治杂志,2015,30(增刊):73-6.
[18]谭红丽,郭英,杨文艳,等.丽江市古城区首次证实一起鼠间鼠疫疫情[J].中国人兽共患病学报,2015,31(11):1086-8.
[2]Hendrix RW.Bacteriophages:evolution of the majiority[J].Theor Popul Biol,2002(61):471-80.
[3]李存香,李伟,王鹏,等.1株温和噬菌体的发现及其流行病学意义[J].医学动物防制,2017,33(10):1073-5.
[4]李存香,王鹏,申小娜,等.野生型鼠疫噬菌体YPO60的分离和生物学特性鉴定[J].中华流行病学杂志,2016,37(6):873-6.
[5]金丽霞,于守鸿,祁芝珍,等.自四川石渠县分离3株野生型鼠疫噬菌体的鉴定[J].中国人兽共患病杂志,2001,17(5):11,124.
[6]宋延富.鼠疫菌以非典型形式在自然界长期保存的研究进展[J].中国地方病防治杂志,1995,10(2):101-3.
[7]张哲,李菁华,逯茵茵,等.粪肠球菌裂解性噬菌体φEn-ZZ8生物学特性的初步分析[J].微生物与感染,2015,10(4):241-6.
[8]张红明,黄庆,罗阳,等.肺炎克雷伯菌噬菌体的分离和生物学特性[J].中华医院感染学杂志,2008,18(5):608-11.
[9]邹玲,任慧英,杨超.一株宽裂解谱大肠埃希菌噬菌体的分离和裂解谱的测定及电镜观察[J].兽医科学,2014,3(5):118-120.
[10]贺雄,王虎.现代鼠疫概论[M].北京:科学出版社,2010:265.
[11]王梅,唐新元,粱莹,等.鼠疫菌基因组差异编码序列的筛选研究[J].中国病原生物学杂志,2017,12(2):115-8.
[12]李存香,靳娟,吴海莲,等.1978-2014年青海省果洛州鼠疫流行病学分析[J].中国病原生物学杂志,2017,12(7):651-3.
[13]潘光合,于建国.鼠疫杆菌在自然生态环境保存状况的研究进展[J].中国国境卫生检疫杂志,2010,33(3):214-6.
[14]何建,靳娟,张青雯,等.一株鼠毒素基因缺失突变鼠疫耶尔森氏菌株的构建和鉴定[J].中国病原生物学杂志,2016,11(6):513-6.
[15]俞东征.鼠疫与假结核耶尔森菌基因组比较分析[J].疾病监测,2011,26(10):760-5.
[16]宋志忠,夏连续,梁云,等.云南玉龙及古城区鼠疫自然疫源地判定及初步研究[J].中国地方病防治杂志,2008,23(1):3-7.
[17]苏丽琼,梁云,吴鹤松,等.云南省2014年鼠疫监测结果[J].中国地方病防治杂志,2015,30(增刊):73-6.
[18]谭红丽,郭英,杨文艳,等.丽江市古城区首次证实一起鼠间鼠疫疫情[J].中国人兽共患病学报,2015,31(11):1086-8.