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Detection of Foodborne Bacterial Pathogens from Individual Filth Flies

There is unanimous consensus that insects are important vectors of foodborne pathogens. However, linking insects as vectors of the pathogen causing a particular foodborne illness outbreak has been challenging. This is because insects are not being aseptically collected as part of an environmental sa...

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Autores principales: Pava-Ripoll, Monica, Pearson, Rachel E.G., Miller, Amy K., Ziobro, George C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354650/
https://www.ncbi.nlm.nih.gov/pubmed/25742168
http://dx.doi.org/10.3791/52372
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author Pava-Ripoll, Monica
Pearson, Rachel E.G.
Miller, Amy K.
Ziobro, George C.
author_facet Pava-Ripoll, Monica
Pearson, Rachel E.G.
Miller, Amy K.
Ziobro, George C.
author_sort Pava-Ripoll, Monica
collection PubMed
description There is unanimous consensus that insects are important vectors of foodborne pathogens. However, linking insects as vectors of the pathogen causing a particular foodborne illness outbreak has been challenging. This is because insects are not being aseptically collected as part of an environmental sampling program during foodborne outbreak investigations and because there is not a standardized method to detect foodborne bacteria from individual insects. To take a step towards solving this problem, we adapted a protocol from a commercially available PCR-based system that detects foodborne pathogens from food and environmental samples, to detect foodborne pathogens from individual flies.Using this standardized protocol, we surveyed 100 wild-caught flies for the presence of Cronobacter spp., Salmonella enterica, and Listeria monocytogenes and demonstrated that it was possible to detect and further isolate these pathogens from the body surface and the alimentary canal of a single fly. Twenty-two percent of the alimentary canals and 8% of the body surfaces from collected wild flies were positive for at least one of the three foodborne pathogens. The prevalence of Cronobacter spp. on either body part of the flies was statistically higher (19%) than the prevalence of S. enterica (7%) and L.monocytogenes (4%). No false positives were observed when detecting S. enterica and L. monocytogenes using this PCR-based system because pure bacterial cultures were obtained from all PCR-positive results. However, pure Cronobacter colonies were not obtained from about 50% of PCR-positive samples, suggesting that the PCR-based detection system for this pathogen cross-reacts with other Enterobacteriaceae present among the highly complex microbiota carried by wild flies. The standardized protocol presented here will allow laboratories to detect bacterial foodborne pathogens from aseptically collected insects, thereby giving public health officials another line of evidence to find out how the food was contaminated when performing foodborne outbreak investigations.
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spelling pubmed-43546502015-03-18 Detection of Foodborne Bacterial Pathogens from Individual Filth Flies Pava-Ripoll, Monica Pearson, Rachel E.G. Miller, Amy K. Ziobro, George C. J Vis Exp Environmental Sciences There is unanimous consensus that insects are important vectors of foodborne pathogens. However, linking insects as vectors of the pathogen causing a particular foodborne illness outbreak has been challenging. This is because insects are not being aseptically collected as part of an environmental sampling program during foodborne outbreak investigations and because there is not a standardized method to detect foodborne bacteria from individual insects. To take a step towards solving this problem, we adapted a protocol from a commercially available PCR-based system that detects foodborne pathogens from food and environmental samples, to detect foodborne pathogens from individual flies.Using this standardized protocol, we surveyed 100 wild-caught flies for the presence of Cronobacter spp., Salmonella enterica, and Listeria monocytogenes and demonstrated that it was possible to detect and further isolate these pathogens from the body surface and the alimentary canal of a single fly. Twenty-two percent of the alimentary canals and 8% of the body surfaces from collected wild flies were positive for at least one of the three foodborne pathogens. The prevalence of Cronobacter spp. on either body part of the flies was statistically higher (19%) than the prevalence of S. enterica (7%) and L.monocytogenes (4%). No false positives were observed when detecting S. enterica and L. monocytogenes using this PCR-based system because pure bacterial cultures were obtained from all PCR-positive results. However, pure Cronobacter colonies were not obtained from about 50% of PCR-positive samples, suggesting that the PCR-based detection system for this pathogen cross-reacts with other Enterobacteriaceae present among the highly complex microbiota carried by wild flies. The standardized protocol presented here will allow laboratories to detect bacterial foodborne pathogens from aseptically collected insects, thereby giving public health officials another line of evidence to find out how the food was contaminated when performing foodborne outbreak investigations. MyJove Corporation 2015-02-13 /pmc/articles/PMC4354650/ /pubmed/25742168 http://dx.doi.org/10.3791/52372 Text en Copyright © 2015, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Environmental Sciences
Pava-Ripoll, Monica
Pearson, Rachel E.G.
Miller, Amy K.
Ziobro, George C.
Detection of Foodborne Bacterial Pathogens from Individual Filth Flies
title Detection of Foodborne Bacterial Pathogens from Individual Filth Flies
title_full Detection of Foodborne Bacterial Pathogens from Individual Filth Flies
title_fullStr Detection of Foodborne Bacterial Pathogens from Individual Filth Flies
title_full_unstemmed Detection of Foodborne Bacterial Pathogens from Individual Filth Flies
title_short Detection of Foodborne Bacterial Pathogens from Individual Filth Flies
title_sort detection of foodborne bacterial pathogens from individual filth flies
topic Environmental Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354650/
https://www.ncbi.nlm.nih.gov/pubmed/25742168
http://dx.doi.org/10.3791/52372
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