Japanese Journal of Food Microbiology
Online ISSN : 1882-5982
Print ISSN : 1340-8267
ISSN-L : 1340-8267
Studies on Methods for Detection and Isolation of Pathogenic Bacteria in Vegetables and Fruits
Enterohemorrhagic Escherichia coli, Salmonella and Listeria
Michiko MIYAHARAKohkichi GOTOHHiroyuki MASAKIAkinobu SAITOSeiji KANEKOTakashi MASUDAHirotaka KONUMA
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2002 Volume 19 Issue 2 Pages 47-55


Several outbreaks of food poisoning have been linked to the consumption of contaminated fresh vegetables and fruits. Enterohemorrhagic E. coli O157 (O157) and Salmonella are the most common causes of these illnesses. Listeria is also supposed to be a cause of food-borne illness. Detection methods for these three pathogenic bacteria from fresh vegetables and fruits were studied by enrichment culture procedure, PCR detection and selective detection. The first step was enrichment by BPW (buffered peptone water) for 20-24 hr at 36°C. After 6 hr of enrichment, 1ml of enrichment solution was transferred to NmEC (EC broth with Novobiocin and bile acid) for separation of O157. After enrichment for 20-24 hr, 1 ml of enrichment solution was treated as a PCR sample. Fewer than 10 cells of O157 or Salmonella per 25 g food samples could be detected by our methods. Listeria (1, 000 cells/25 g) were also detectable by these methods. Criteria for the multiplex PCR (O157, Salmonella and Listeria) were key factors for further testing (Limitations of No. detected were2.3×104, 2.7×105 and2.2×105 cfu/ml, respectively). Selective cultures were used when PCR was positive for pathogenic bacteria.
These methods are economical and effective for detection of these pathogenic bacteria from fresh vegetables and fruits.

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