Abstract
Foodborne parasites (FBP) are of major public health importance and warrant appropriate detection and control strategies. Most of the FBP considered for risk-ranking by a panel of experts are potentially transmitted via consumption of contaminated fresh produce, including berries. In this study we focused on the potential of three FBP, namely Echinococcus multilocularis, Toxoplamsa gondii, and Cyclospora cayetanensis, as contaminants of berries. Surveys to assess these parasites as contaminants of fresh produce in general, and berries in particular, are scanty or non-existent mainly due to the lack of optimized laboratory methods for detection. The aim of the present study was to develop and evaluate a novel multiplex qPCR for the simultaneous detection of E. multilocularis, T. gondii, and C. cayetanensis from berry fruits. The efficiency and linearity of each channel in the multiplex qPCR were within the acceptable limits for the range of concentrations tested. Furthermore, the method was shown to have good repeatability (standard deviation ≤0.2 Cq) and intermediate precision (pooled standard deviation of 0.3-0.6 Cq). The limit of detection was estimated to 10 oocysts for Toxoplasma and Cyclospora, and 5 eggs for Echinococcus per 30 g of raspberries or blueberries. In conclusion, evaluation of the present method showed that the newly developed multiplex qPCR is highly specific, precise, and robust method that has potential for application in food-testing laboratories.
Highlights
Food safety is a major concern for the global community, with an estimated 600 million cases of foodborne illnesses occurring annually (FAO/ WHO, 2019)
The triplex qPCR showed good efficiency and linearity for the range of concentrations tested for the three parasites, with assay efficiency of 104%, 92%, and 105%, for Echinococcus, Toxoplasma, and Cyclospora, respectively
The assay was designed for analysis of berry fruits, due to associations in the literature of these parasites with berries ( Cyclospora and E. multilocularis), it could be perfectly well used with other types of fresh produce that could act as vehicles of infection, such as salad vegetables
Summary
Food safety is a major concern for the global community, with an estimated 600 million cases of foodborne illnesses occurring annually (FAO/ WHO, 2019). Foodborne illnesses could be due to infection (biological risk) or toxicity (chemical risk) resulting from consumption of contaminated food. The infection could result from consuming food contaminated with parasites, bacteria, or viruses. This is true for foods that are consumed fresh and raw, such as fruits and vegetables. Toxoplasmosis, trichinellosis, cystic echinococcosis, and cryptosporidiosis are diseases caused by those parasites listed as ‘top five’ priority (Bouwknegt et al, 2018). Among these five prioritized parasites, four can be transmitted via contaminated fresh produce. Cyclospora cayetanensis was not ranked among the top five in Europe, we chose to include it due to the many recent outbreaks, in USA, in which transmission via contaminated fresh produce has been implicated
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
