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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Parasitic Diseases Laboratory » Research » Publications at this Location » Publication #319804
Title: Strategy for simultaneous molecular detection of the protozoan parasites Toxoplasma, Cryptosporidium and Giardia in food matrices and persistence on leaves of vegetables during storage at 4°C

Author
item HOHWEYWE, JEANNE - Universite De Reims Champagne-Ardenne
item CAZEAUX, CATHERINE - Actalia Securite Des Aliment
item TRAVAILE, EMMANUELLE - Desiderio Finamore Veterinary Research Institute (FEPAGRO)
item DUMETRE, AURELIEN - Aix-Marseille University
item AUBERT, DOMINIQUE - Universite De Reims Champagne-Ardenne
item LANGUET, EMILIE - Actalia Securite Des Aliment
item TERRYN, CHRISTINE - Universite De Reims Champagne-Ardenne
item Dubey, Jitender
item AZAS, NADINE - Aix-Marseille University
item HOUSSIN, MARYLINE - Desiderio Finamore Veterinary Research Institute (FEPAGRO)
item LOIC, FAVENNEC - University Of Rouen
item VILLENA, ISABELLE - Hopital Maison Blanche
item LA CARBONA, STEPHANIE - Actalia Securite Des Aliment

Submitted to: Food Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/12/2016
Publication Date: 1/12/2016
Citation: Hohweywe, J., Cazeaux, C., Travaile, E., Dumetre, A., Aubert, D., Languet, E., Terryn, C., Dubey, J.P., Azas, N., Houssin, M., Loic, F., Villena, I., La Carbona, S. 2016. Strategy for simultaneous molecular detection of the protozoan parasites Toxoplasma, Cryptosporidium and Giardia in food matrices and persistence on leaves of vegetables during storage at 4°C. Food Microbiology. 57:36-44.

Interpretive Summary: Toxoplasmosis, caused by the single celled parasite, Toxoplasma gondii, continues to be a public health problem worldwide. This parasites infects all warm-blooded hosts, including humans. It causes mental retardation and loss of vision in children, and abortion in livestock. The ingestion of food and water contaminated with resistant stage of the parasite, the oocyst, is now considered a major mode of transmission of this parasite. Of all the hosts infected, only cats are known to excrete oocysts in feces. Cats can excrete millions of oocysts after eating an infected prey, such as a mouse or a bird. Oocysts can survive outdoors for months. Currently, there is no efficient method of detection of oocysts in contaminated fruits or vegetables. In the present papers the authors report on magnetic capture PCR method to detect oocysts on fruits. The results will be of interests to biologists, parasitologists, and public health workers.

Technical Abstract: Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis are emerging pathogen parasites in the food domain. However, without standardized method for their detection in food matrices, parasitic foodborne outbreaks remain neglected. In this study, a new immunomagnetic separation assay (IMS Toxo) targeting the oocyst’s wall of T. gondii was developed using a specific purified monoclonal antibody. Performance of this IMS Toxo coupled to microscopic and qPCR analyses was evaluated in terms of limit of detection (LOD) and recovery rate (RR) on: i) simple matrix (LOD = 5 oocysts; RR between 5 and 56%); ii) raspberries and basil (LOD = 33 oocysts/g; RR between 0.2 and 35%). To simultaneously extract the three protozoa from these food matrices, IMS of Cryptosporidium and Giardia (oo)cysts was finally combined to T. gondii oocysts concentration without IMS Toxo. This strategy associated to qPCR detection led to LOD < 1 to 3 (oo)cysts/g and recovery rates between 2 and 35%. This procedure was coupled to RT-qPCR analyses and showed that the three protozoa persisted onto the surface of leaves of basil and remained viable following storage at 4 °C for 8 days. These data strengthen the need to consider these protozoa in food safety.