病原性副溶血弧菌在天然海水中的饥饿耐受研究
Starvation responses of pathogenic Vibrio parahaemolyticus in natural Seawater
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摘要: 将副溶血弧菌培养至对数生长期,用天然海水洗脱,调整菌浓度至107个/cm3左右后置于27℃恒温培养箱进行饥饿试验.结果表明:饥饿初期细菌总数、可培养菌数和活菌数都有较大幅度上升;饥饿中、后期,总菌数与活菌数缓慢下降,而可培养菌数下降速度较快.饥饿前副溶血弧菌呈短杆状,染色均匀;饥饿后许多细胞中央出现染色较浅的区域,细胞形态变为椭圆形.副溶血弧菌对大黄鱼表皮黏液的黏附量随着饥饿时间延长而急剧下降,饥饿7 d后的黏附量接近于空白.用SDS-PAGE分析不同饥饿时期菌体蛋白质的差异,结果表明:饥饿30 d后菌体蛋白条带比饥饿前的蛋白质条带少;饥饿60 d后菌体蛋白质条带比饥饿30 d时的多,但比饥饿前少.研究结果对于揭示副溶血弧菌的流行病学特征有重要的参考价值.Abstract: Pathogenic V.parahaemolyticus cells in log-phase were collected and the concentration was adjusted to 107 cells/cm3 in natural sea water for starvation studies.The results showed that: total bacteria number、viable bacteria number and CFU number of V.parahaemolyticus increased remarkably at the initial starvation stages;total bacteria number、viable bacteria number and CFU number fell at metaphase and anaphase,among them,the total bacteria number and viable bacteria number fell slowly while the CFU number fell more quickly.Before starvation,the V.parahaemolyticus cells were shortrods and dyed equably;after starvation,a light area appeared at the cell centre and some cells turned into oval.The amount of V.parahaemolyticus cells adhered to the large yellow croaker epidermal mucus dropped sharply during starvation,approaching the level of the control after 7 days starvation.The cellular protein of V.parahaemolyticus was extracted and analyzed by SDS-PAGE,the results showed that: bacteria starved for 30 d had less protein bands in the gel than the unstarved ones;bacteria starved for 60 d had more protein bands compared to those starved for 30 d,but less than the unstarved ones.The results obtained were useful to the underst anding of the epidemiologic characteristics of V.parahaemolyticus.
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Key words:
- Vibrio parahaemolyticus /
- starvation /
- survival /
- adhesion
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MORITA R Y.The starvation survival state of microorganism in nature and its relationship to the bioavailable energy[J].Experientia,1990,46(8):813-817. OLIVER J D,NILSSON L,JELLEBERG K.Formation of nonculturable Vibrio vulnificus cells and its relationship to the starvation state[J].Appl Environ Microbiol,1991,57(9):2 640-2 644. PALUDAN-MULLER C,WEICHART D,MCDOUGALD D,et al.Analysis of starvation conditions that allow for prolonged culturability of Vibrio vulnificus at low temperature[J].Microbiology,1996,142:1 675-1 684. OLIVER J D.Formation of viable but nonculturable cells[A].KJELLEBERG S.Starvation in bacteria[M].New York:Plenum Press,1993.239-272. JOHNSTON M D,BROWN M H.An investigation into the changed physiological state of Vibrio bacteria as a survival mechanism in response to cold temperatures and studies on their sensitivity to heating and freezing[J].Journal of Applied Microbiology,2002,92(6):1 066-1 077. JIANG X P,CHAI T J.Survival of Vibrio parahaemolyticus at Low Temperatures under Starvation Conditions and Subsequent Resuscitation of Viable,Nonculturable Cells[J].Appl Environ Microbiol,1996,62(4):1 300-1 305 鄢庆枇,王军,苏永全,等.网箱养殖大黄鱼弧菌病研究[J].集美大学学报(自然科学版),2001,6(3):191-196. 金珊,王国良,赵青松,等.海水网箱养殖大黄鱼弧菌病的流行病学研究[J].水产科学,2005,24(1):17-19. JOSEPH S W,COLWELL R R,KAPER J B.Vibrio parahaemolyticus and related halophilic vibrios[J].Crit Rev Microbiol,1982,10(1):77-124. COLWELL R R,RAVTON P R,GRIMES D J,et al.Viable but non-culturable Vibrio cholerae and related pathogens in the environment:implications for release of genetically engineered microorganisms[J].Bio/Technology,1985,3:817-820. LINDER K,OLIVER J D.Membrane fatty acid and virulence changes in the viable but nonculturable state of Vibrio vulnificus[J].Appl Environ Microbiol,1989,55(1):2 837-2 842. WONG H C,WANG P.Induction of viable but nonculturable state in Vibrio parahaemolyticus and its susceptibility to environmental stresses[J].Journal of Applied Microbiology,2004,96(2):359-366. FURUYA T,KAMADA T,MURAKAMI T,et al.Laser scanning cytometry allows detection of cell death with morphological features of apoptosis in cells stained with PI[J].Cytometry,1998,29(2):173. BORDAS M A,BALEBONA M C,ZORRILA I,et al.Kinetics of adhesion of selected fish-pathogenic Vibrio strains to skin mucus of gilt-head sea bream(Sparus aurata L.)[J].Appl Environ Microbiol,1996,62:3 650-3 654. MARSHAK D R,KADONAGA J,BURGESS R R,et al.Strategies for Purification and Characterization:A Laboratory Course Manual[M].New York:Cold Spring Harbor Laboratory Press,1996.249-256. FERNANDEZ BOTRAN R,VETVCKA V.Methods in cellular immunology[M].New York:CRC Press,1995.29-46. MORITA R Y.Bioavailability of energy and the starvation state[A].KJELLEBERG S.Starvation in Bacteria[M].New York:Plenum Press,1993.1-23. NILSSON L,OLIVER J D,KJELLEBERG S.Resuscitation of Vibrio vulnificus from the viable but nonculturable state[J].Bacteriol,1991,173(16):5 054-5 059. JOHN A,JEFF M,ARMINE S.A rapid microplate method for quantifying inhibition of bacterial adhesion to eukaryotic cells[J].Journal of Microbiological Methods,2005,60 (1):55-62. CHABRILLON M,BORDAS M A,MORINIGO M A,et al.Kinetics of adhesion of Listonella anguillarum to the mucus of gilt-head seabream,and the implication of surface components[J].Aquaculture Research,2004,35 (4):403-409. BALEBONA M C,MORINIGO M A,FARIS A,et al.Influence of salinity and pH on the adhesion of pathogenic Vibrio strains to Sparus aurata skin mucus[J].Aquaculture,1995,132:113-120. PAYNE G A,NYSTROM G J,BHATNAGAR D,et al.Cloning of the afl-2 gene involved in aflatoxin biosynthesis from Aspergillus flavus[J].Appl Environ Microbiol,1993,59(1):156-162. NYSTROM T,FLARDH K,KJELLEBERG S.Responses to multiple nutrient starvation in marine Vibrio sp.Strain CCUG 15956[J].Bacteriol,1990,172(12):7 085-7 097. KOLTER R,SIEGELE D A,TORMO A.The stationary phase of the bactrial life cycle[J].Ann Rev Microbiol.1993,17:855-874. MORTON D S,LIVER J D.Induction of carbon starvation-induced proteins in Vibrio vulnificus[J].Appl Environ Microbiol,1994,60(10):3 653-3 659.
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