Effect of environmental stresses on antibody-based detection of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis and Listeria monocytogenes.
OBJECTIVE
To study the reaction patterns of selected antibodies to Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis and Listeria monocytogenes cells exposed to various environmental stresses.


RESULTS
Escherichia coli O157:H7, Salmonella Enteritidis and L. monocytogenes cells subjected to different environmental stress of temperatures (4 and 45 degrees C), NaCl (5.5%), oxidative stress (15 mmol(-1) H2O2), acidic pH (5.5) and ethanol (5%) for 3 h (short-term stress) or for 5 days (long-term stress) were analysed by ELISA and Western blotting. The ELISA results indicated that most stresses caused 12-16% reductions in reaction for anti-E. coli O157:H7 and 20-48% reductions for anti-Salmonella polyclonal antibodies during short-term stress, whereas the most stresses exhibited enhanced reaction (44-100% increase) with the anti-L. monocytogenes polyclonal antibody. During long-term stress exposure to combined stress conditions of pH 5.5, 3.5% NaCl at 12 degrees C or at 4 degrees C, antibody reactions to the three pathogens were highly variable with the combined stress at 4 degrees C showing the most reductions (8-40%). Likewise, there were about 18-59% reductions in antibody reactions with pathogens when cultured in hotdog samples with the combined stress conditions. Western blot analyses of crude cell surface antigens from both short- and long-term stressed cells revealed that the changes in antibody reactions observed in ELISA were either because of repression, expression or possible denaturation of antigens on the surface of cells.


CONCLUSIONS
Overall, the antibody reactions were significantly reduced in pathogens exposed to both short- and long-term environmental stresses in culture medium or in meat sample because of expression, repression or denaturation of specific antigens in cells.


CONCLUSIONS
In order to ensure the reliable detection of foodborne pathogens using antibody-based methods, the influence of stress on antibody reactions should be thoroughly examined and understood first as the physiological activities in cells are often altered in response to a stress.
Hahm
B-K
BK
Department of Food Science, Molecular Food Microbiology Laboratory, Purdue University, West Lafayette, IN 47907-2009, USA.
Bhunia
A K
AK
D000907:Antibodies, Bacterial / Q000032:analysis*; D001419:Bacteria / Q000145:classification / Q000302:isolation & purification*; D015153:Blotting, Western / Q000379:methods; D004777:Environment; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D019453:Escherichia coli O157 / Q000276:immunology / Q000302:isolation & purification; D005516:Food Microbiology; D006801:Humans; D008089:Listeria monocytogenes / Q000276:immunology / Q000302:isolation & purification; D008461:Meat Products / Q000382:microbiology; D018384:Oxidative Stress / Q000502:physiology; D012477:Salmonella enteritidis / Q000276:immunology / Q000302:isolation & purification
D000907:Antibodies, Bacterial