金黄色葡萄球菌噬菌体的生物学特性及其在牛奶中的抑菌应用
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摘要
分离纯化一株裂解性金黄色葡萄球菌(Staphylococcus aureus)噬菌体,鉴定其理化和生物学性质,并研究噬菌体在牛奶中的抑菌作用。本研究以金黄色葡萄球菌(ATCC6538)为宿主菌,采用点滴法和双层平板法,分离获得特异性针对金黄色葡萄球菌的具有较高裂解活性的噬菌体,命名为qdsa001。经过透射电镜观察表明,该噬菌体属于长尾噬菌体科(Siphoviridae),最佳感染复数为0.1。一步生长曲线结果显示噬菌体感染宿主菌的潜伏期约为10min,爆发期约90min,平均裂解量约为32.2。它对60℃以下的温度耐受力较强,最适pH值为5～9左右。同时,牛奶中抑菌应用实验表明,噬菌体qdsa001对牛奶中金黄色葡萄球菌的生长具有良好的抑制作用。
By the combined use of drop method and double-layer plate method,a phage with specific lytic activity against Staphylococcus aureus was isoalted from urban sewage and named as qdsa001.The host strain used was Staphylococcus aureus ATCC 6538.Some physiochemical and biological characteristics of the pahge qdsa 001 were identified.Besides,its antibacterial effect in milk was also investigated.Transmission electron microscopic(TEM) observation showed that qdsa 001 belonged to the Siphoviridae family.The phage had a strong endurance to temperature below 60 ℃,and its optimal pH and MOI(multiplicity of infection) were approximately 7-8 and 0.1,respectively.The one-step growth curve plot showed that the latent period and lytic period of this phage were 10 min and 90 min,respectively.In addition,the average burst size was 32.2.Therefore,phage qdsa 001 is a virulent bacteriophage and can effectively inactivate Staphylococcus aureus in ready-to-eat milk.
By the combined use of drop method and double-layer plate method,a phage with specific lytic activity against Staphylococcus aureus was isoalted from urban sewage and named as qdsa001.The host strain used was Staphylococcus aureus ATCC 6538.Some physiochemical and biological characteristics of the pahge qdsa 001 were identified.Besides,its antibacterial effect in milk was also investigated.Transmission electron microscopic(TEM) observation showed that qdsa 001 belonged to the Siphoviridae family.The phage had a strong endurance to temperature below 60 ℃,and its optimal pH and MOI(multiplicity of infection) were approximately 7-8 and 0.1,respectively.The one-step growth curve plot showed that the latent period and lytic period of this phage were 10 min and 90 min,respectively.In addition,the average burst size was 32.2.Therefore,phage qdsa 001 is a virulent bacteriophage and can effectively inactivate Staphylococcus aureus in ready-to-eat milk.
引文
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[18]李波.原料奶中病原菌的调查及代表性病原菌的研究[J].食品工程,2007(3):3-6.
[2]GUPTA R,PRASAD Y.P-27/HP endolysin as antibacterial agentfor antibiotic resistant Staphylococcus aureus of human infections[J].Current Microbiology,2011,63(1):39-45.
[3]LOWY F D.Staphylococcal aureus infections[J].N Engl J Med,1998,339:520-532.
[4]刘秀梅.食源性疾病监控技术的研究[J].中国食品卫生杂志,2004,16(1):3-9.
[5]巢国祥,焦新安,周丽萍,等.食源性金黄色葡萄球菌流行特征、产肠毒素特性及耐药性研[J].中国卫生检验杂志,2006,16(8):904-907.
[6]黄冰,邓志爱,谭铭雄,等.食品中金黄色葡萄球菌污染状况、产肠毒素特性及耐药性研究[J].中国卫生检验杂志,2009,19(6):1380-1382.
[7]HAGENS S,LOESSNER M J.Bacteriophage for biocontrol offoodborne pathogens:calculations and considerations[J].CurrentPharmaceutical Biotechnology,2010,11(1):58-68.
[8]刘伟,王学琴,董世雷,等.噬菌体及其内溶素的应用研究进展[J].浙江农业学报,2010,22(5):702-708.
[9]CARLTONRM,NOORDMANWH,BISWASB,etal.Bacteriophage P100 for control of Listeria monocytogenes in foods:genome sequence,bioinformatic analyses,oral toxicity study,andapplication[J].Regulatory Toxicology and Pharmacology,2005,43(3):301-312.
[10]杜崇涛.大肠杆菌O157噬菌体的分离鉴定及其初步应用[D].长春:吉林大学,2008.
[11]LU Z,BREIDT F J,FLEMING H P.Isolation and characterizationof a Lactobacillus plantarum bacteriophageΦJL21 from a cucumberfermentation[J].Food Microbiol,2003,84(2):225-235.
[12]MARTíNEZ B,OBESO J M,RODRíGUEZ A,et al.Nisin-bacteriophage crossresistance in Staphylococcus aureus[J].International Journal of Food Microbiology,2008,122(3):253-258.
[13]洪健,周雪平.ICTV第八次报告的最新病毒分类系统[J].中国病毒学,2006,21(1):84-96.
[14]于龙,温占波,杨文慧,等.一株粘质沙雷氏菌烈性噬菌体污水分离及特性[J].微生物学报,2008,48(4):498-502.
[15]HENDRIX R W.Bacteriophage genomics[J].Curr Opin Microbiol,2003,6(5):506-511.
[16]石琰璟,郭清吉,葛晓萍,等.金黄色葡萄球菌噬菌体的分离纯化[J].青岛科技大学学报,2007,28(3):205-207.
[17]李陇平,张智英.金黄色葡萄球菌烈性噬菌体的分离鉴定和最佳保存方法研究[J].中国畜牧兽医,2011,38(6):141-146.
[18]李波.原料奶中病原菌的调查及代表性病原菌的研究[J].食品工程,2007(3):3-6.