Browse > Article

Improved Detection of ${\gamma}-Irradiated$ Vibrio vulnificus after Heat and Cold Shock Treatment by Using Ethidium Monoazide Real-time PCR  

Lee, Jung-Lim (Department of Food Science, Massachusetts Agricultural Experiment Station, University of Massachusetts)
Levin, Robert E. (Department of Food Science, Massachusetts Agricultural Experiment Station, University of Massachusetts)
Publication Information
Food Science and Biotechnology / v.18, no.3, 2009 , pp. 788-792 More about this Journal
Abstract
Gamma $({\gamma})-irradiation$ can be used to control pathogens such as Vibrio vulnificus in seafood. The effects of irradiation on microbial cell populations (%) have been studied in order to develop detection methods for irradiated foods. The method used in this study was ethidium bromide monoazide (EMA) real-time polymerase chain reaction (PCR), using V. vulnificus specific primer, EMA, and $SYBR^{(R)}$ Green to discriminate between ${\gamma}-irradiated$ and non-irradiated cells. Confocal microscope examination showed that ${\gamma}-irradiation$ damaged portions of the cell membrane, allowing EMA to penetrate cells of irradidated V. vulnificus. ${\gamma}-Irradiation$ at 1.08 KGy resulted in log reduction ($-1.15{\pm}0.13$ log reduction) in genomic targets derived from EMA real-time PCR. The combination cold/heat shock resulted in the highest ($-1.74{\pm}0.1$ log reduction) discrimination of dead irradiated V. vulnificus by EMA real-time PCR.
Keywords
$({\gamma})-irradiation$; ethidium bromide monoazide; real-time polymerase chain reaction (PCR); Vibrio vulnificus; cold and heat shock;
Citations & Related Records

Times Cited By Web Of Science : 0  (Related Records In Web of Science)
연도 인용수 순위
  • Reference
1 Hsu WY, Wei CI, Tamplin ML. Enhanced broth media for selective growth of Vibrio vulnificus. Appl. Environ. Microb. 64: 2701-2704 (1998)   PUBMED   ScienceOn
2 Blake PA, Merson MH, Weaver RE, Hollis DG., Heublen PC. Disease caused by a marine Vibrio: Clinical characteristics and epidemiology. New Engl. J. Med. 300: 1-5 (1979)   DOI   ScienceOn
3 Bhide MR, Paturkar AM, Sherikar AT, Waskar VS. Presensitization of microorganisms by acid treatment to low dose gamma irradiation with special reference to Bacillus cereus. Meat. Sci. 58: 253-258 (2001)   DOI   ScienceOn
4 Villavicencio ALCH, Araujo MM, Marin-Huachacaa NS, Mancini-Fiho J, Delincee H. Identification of irradiated refrigerated poultry with the DNA comet assay. Radiat. Phys. Chem. 71: 189-191 (2004)   DOI   ScienceOn
5 Araujo MM, Marin-Huachacaa NS, Mancini-Fiho J, Delincee H, Villavicencio ALCH. Identification of irradiated refrigerated pork with the DNA comet assay. Radiat. Phys. Chem. 71: 185-187 (2004)   DOI   ScienceOn
6 Marin-Huachacaa N, Delincee H, Mancini-Fiho J, Villavicencio ALCH. Use of the DNA comet assay to detect beef meat treated by ionizing radiation. Meat Sci. 71: 446-450 (2005)   DOI   ScienceOn
7 Nogva HK, Dromtorp SM, Nissen H, Rudi K. Ethidium monoazide for DNA-based differentiation of viable and dead bacteria by 5'-nuclease PCR. Biotechniques 34: 804-813 (2003)   PUBMED   ScienceOn
8 Abolmatty A, Vu C, Oliver J, Levin RE. Development of a new lysis solution for releasing genomic DNA from bacterial cells for DNA amplification by polymerase chain reaction. Microbios 101: 181-189 (2000)   PUBMED   ScienceOn
9 Larsen JL. Vibrio anguillarum: Influence of temperature, pH, NaCl concentration, and incubation time on growth. J. Appl. Bacteriol. 57: 237-246 (1984)   DOI
10 Calcott PH, MacLeod RA. The survival of Escherichia coli from freeze-thaw damage: The relative importance of wall and membrane damage. Can. J. Microbiol. 21: 1960-1968 (1975)   DOI   ScienceOn
11 Oh KN, Lee SY, Lee HJ, Kim KE, Yang JS. Screening of gamma irradiation spices in Korea by using a microbiological method (DEFT/APC). Food. Control 14: 489-494 (2003)   DOI   ScienceOn
12 Tamplin ML, Rodrick GE, Blake NJ, Cuba T. Isolation and characterization of Vibrio vulnificus from two Florida estuaries. Appl. Environ. Microb. 44: 1466-1470 (1982)   PUBMED   ScienceOn
13 Panicker G, Myers ML, Bej AK. Rapid detection of Vibrio vulnificus in shellfish and gulf of Mexico water by real-time PCR. Appl. Environ. Microb. 70: 498-507 (2004)   DOI   ScienceOn
14 Fujiwara-Nagata E, Kogure K, Kita-Tsukamoto K, Wada M, Eguchi M. Characteristics of Na+-dependent respiratory chain in Vibrio anguillarum, a fish pathogen, in comparison with other marine Vibrios. FEMS Microbiol. Ecol. 44: 225-230 (2003)   DOI   ScienceOn
15 Levin RE. The application of real-time PCR to food and agricultural systems. A review. Food Biotechnol. 18: 97-133 (2004)   DOI   ScienceOn
16 Lee JL, Levin RE. Use of ethidium bromide monoazide for quantification of viable and dead mixed bacterial flora from fish fillets by polymerase chain reaction. J. Microbiol. Meth. 67: 456-462 (2006)   DOI   ScienceOn
17 Lee JL, Levin RE. Discrimination of $\gamma$-irradiated Vibrio vulnificus by using ethidium monoazide real-time polymerase chain reaction. J. Appl. Microbiol. 104: 728-734 (2008)   DOI   ScienceOn
18 Bennett GM, Seaver A, Calcott PH. Effect of defined lipopolysaccharide core defects on resistance of salmonella typhimurium to freezing and thawing and other stresses. Appl. Environ. Microb. 42: 843-849 (1981)   PUBMED   ScienceOn
19 Ray B, Speck ML. Freezing-injury in bacteria. Crit. Rev. Cl. Lab. Sci. 4: 161-213 (1973)   DOI   ScienceOn
20 Diehl JF. Safety of Irradiated Foods. Marcel Dekker, Inc., New York, NY, USA. pp. 95-144 (1990)