title: Simultaneous detection of the protozoan parasites Toxoplasma, Cryptosporidium and Giardia in food matrices and their persistence on basil leaves.
abstract: Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis are emerging pathogen parasites in the food domain. However, without standardized methods 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%). Finally, to simultaneously extract the three protozoa from these food matrices, T. gondii oocysts were directly concentrated (without IMS Toxo) from the supernatant of the IMS of Cryptosporidium and Giardia (oo)cysts. This strategy associated to qPCR detection led to LOD <1 to 3 (oo)cysts/g and RR between 2 and 35%. This procedure was coupled to RT-qPCR analyses and showed that the three protozoa persisted on the 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.
authors.affiliation: EA 3800, Protozooses transmises par l'alimentation, Laboratoire de Parasitologie Mycologie, Université de Reims Champagne Ardenne, Faculté de Médecine, SFR Cap Santé Fed 4231, 51 rue Cognacq Jay, 51096 Reims, France. Electronic address: jeanne.hohweyer@gmail.com.
authors.forename: Jeanne
authors.initials: J
authors.lastname: Hohweyer
authors.affiliation: ACTALIA Food Safety Department, 310 rue Popielujko, 50000 Saint-Lô, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Cazeaux
authors.affiliation: Labéo Frank Duncombe, 1 route de Rosel, Saint Contest, 14053 Caen Cedex 4, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Travaillé
authors.affiliation: ACTALIA Food Safety Department, 310 rue Popielujko, 50000 Saint-Lô, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Languet
authors.affiliation: Infections Parasitaires, Transmission, Physiopathologie et Thérapeutique, UMR MD3, Faculté de Pharmacie, Aix Marseille Université, 27 Boulevard Jean Moulin, CS 30064, 13385 Marseille Cedex 05, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Dumètre
authors.affiliation: EA 3800, Protozooses transmises par l'alimentation, Laboratoire de Parasitologie Mycologie, Université de Reims Champagne Ardenne, Faculté de Médecine, SFR Cap Santé Fed 4231, 51 rue Cognacq Jay, 51096 Reims, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Aubert
authors.affiliation: Plateforme Imagerie Cellulaire et Tissulaire, Université de Reims Champagne Ardenne, 51 rue Cognacq Jay, 51095 Reims, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Terryn
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Animal Parasitic Diseases Laboratory, Building 1001, Beltsville, MD 20705 2350, USA.
authors.forename: Jitender P
authors.initials: JP
authors.lastname: Dubey
authors.affiliation: Infections Parasitaires, Transmission, Physiopathologie et Thérapeutique, UMR MD3, Faculté de Pharmacie, Aix Marseille Université, 27 Boulevard Jean Moulin, CS 30064, 13385 Marseille Cedex 05, France.
authors.forename: Nadine
authors.initials: N
authors.lastname: Azas
authors.affiliation: Labéo Frank Duncombe, 1 route de Rosel, Saint Contest, 14053 Caen Cedex 4, France.
authors.forename: Maryline
authors.initials: M
authors.lastname: Houssin
authors.affiliation: EA 3800, Protozooses transmises par l'alimentation Laboratoire de Parasitologie Mycologie, Université de Rouen, 76183 Rouen Cedex, France.
authors.forename: Favennec
authors.initials: F
authors.lastname: Loïc
authors.affiliation: EA 3800, Protozooses transmises par l'alimentation, Laboratoire de Parasitologie Mycologie, Université de Reims Champagne Ardenne, Faculté de Médecine, SFR Cap Santé Fed 4231, 51 rue Cognacq Jay, 51096 Reims, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Villena
authors.affiliation: ACTALIA Food Safety Department, 310 rue Popielujko, 50000 Saint-Lô, France. Electronic address: s.lacarbona@actalia.eu.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: La Carbona
mesh_terms: D003458:Cryptosporidium / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis; D005872:Giardia / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D018649:Ocimum basilicum / Q000469:parasitology*; D033661:Oocysts / Q000254:growth & development; D018515:Plant Leaves / Q000469:parasitology; D065856:Rubus / Q000469:parasitology*; D014122:Toxoplasma / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*
keywords: Fruits; Immunomagnetic separation; Microscopy; Persistence; Protozoa; Real-time qPCR; Vegetables; Viability