Evaluation of real-time qPCR-based methods to detect the DNA of the three protozoan parasites Cryptosporidium parvum, Giardia duodenalis and Toxoplasma gondii in the tissue and hemolymph of blue mussels (M. edulis)

Abstract Background: The association between the parasitic illnesses and the consumption of contaminated food is more and more described. However, there is still a lack of studies investigating the occurrence of parasitic contamination in food matrices. The aim of the present study was to assess the presence of T. gondii and Cryptosporidium spp. oocysts and G. duodenalis cysts, in three leafy greens (coriander, lettuce and parsley) commonly consumed raw.Methods: A total of 152 leafy green samples were collected in Marrakech from April 2018 to October 2019. Parasites were eluted and concentrated before detection of their DNA by real-time qPCR. Results: The analysis revealed an overall rate of contamination of 32.2% (49/152), with 29.6% (45/152) positive for T. gondii, 2.6% (4/152) for G. duodenalis, while Cryptosporidium spp was not detected. Conclusion: The results showed that leafy greens vegetables available in markets of Morocco are subjected to protozoan parasites contaminations. Thus, humans can be exposed to these parasites through vegetables consumption. Further investigations can be performed to acquire new epidemiological data on the health risk of these protozoan diseases in Morocco.

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Assessing viability and infectivity of foodborne and waterborne stages (cysts/oocysts) of Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii: a review of methods

Parasite ◽

10.1051/parasite/2018009 ◽

2018 ◽

Vol 25 ◽

pp. 14 ◽

Cited By ~ 39

Author(s):

Angélique Rousseau ◽

Stéphanie La Carbona ◽

Aurélien Dumètre ◽

Lucy J. Robertson ◽

Gilles Gargala ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Exposure Assessment ◽

Foodborne Pathogens ◽

Giardia Duodenalis ◽

Human Infection ◽

Control Measures ◽

World Health ◽

Protozoan Parasites ◽

Analytical Strategy ◽

Health Organization

Giardia duodenalis, Cryptosporidium spp. and Toxoplasma gondii are protozoan parasites that have been highlighted as emerging foodborne pathogens by the Food and Agriculture Organization of the United Nations and the World Health Organization. According to the European Food Safety Authority, 4786 foodborne and waterborne outbreaks were reported in Europe in 2016, of which 0.4% were attributed to parasites including Cryptosporidium, Giardia and Trichinella. Until 2016, no standardized methods were available to detect Giardia, Cryptosporidium and Toxoplasma (oo)cysts in food. Therefore, no regulation exists regarding these biohazards. Nevertheless, considering their low infective dose, ingestion of foodstuffs contaminated by low quantities of these three parasites can lead to human infection. To evaluate the risk of protozoan parasites in food, efforts must be made towards exposure assessment to estimate the contamination along the food chain, from raw products to consumers. This requires determining: (i) the occurrence of infective protozoan (oo)cysts in foods, and (ii) the efficacy of control measures to eliminate this contamination. In order to conduct such assessments, methods for identification of viable (i.e. live) and infective parasites are required. This review describes the methods currently available to evaluate infectivity and viability of G. duodenalis cysts, Cryptosporidium spp. and T. gondii oocysts, and their potential for application in exposure assessment to determine the presence of the infective protozoa and/or to characterize the efficacy of control measures. Advantages and limits of each method are highlighted and an analytical strategy is proposed to assess exposure to these protozoa.

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A cloth-based hybridization array system for rapid detection of the food- and waterborne protozoan parasites Giardia duodenalis, Cryptosporidium spp. and Toxoplasma gondii

Food and Waterborne Parasitology ◽

10.1016/j.fawpar.2021.e00130 ◽

2021 ◽

pp. e00130

Author(s):

Sarah J. Reiling ◽

Harriet Merks ◽

Shawna Zhu ◽

Ryan Boone ◽

Nathalie Corneau ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Rapid Detection ◽

Giardia Duodenalis ◽

Protozoan Parasites ◽

Cryptosporidium Spp

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Cryptosporidium spp., Giardia duodenalis and Toxoplasma gondii detection in fresh vegetables consumed in Marrakech, Morocco

African Health Sciences ◽

10.4314/ahs.v20i4.19 ◽

2020 ◽

Vol 20 (4) ◽

pp. 1669-78

Author(s):

Salma Berrouch ◽

Sandie Escotte-Binet ◽

Yassine Amraouza ◽

Pierre Flori ◽

Dominique Aubert ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Giardia Duodenalis ◽

Epidemiological Data ◽

Contaminated Water ◽

Protozoan Parasites ◽

Leafy Greens ◽

Cryptosporidium Spp ◽

Vegetable Samples ◽

Fresh Vegetables

Background: Protozoan parasites such as Toxoplasma gondii, Giardia duodenalis, and Cryptosporidium spp., can be transmitted to humans via accidental consumption of contaminated water, fresh produce and foodstuffs. There is a lack of epidemiological data about these pathogens in Morocco. Hence the aim of this study, which is the determination of their prevalence in some leafy greens and root vegetables sold in Marrakech. Methods: A total of 132 vegetable samples including carrot, coriander, lettuce, parsley and radish were purchased monthly from three different markets in Marrakech from March 2017 to January 2018, pre-treated and subjected to microscopic and molecular analyses. Results: Of the 132 samples of vegetables analyzed by qPCR, the overall rate of protozoan was 21.21% (28/132); 22 samples were found to be contaminated with T. gondii, 6 with G. duodenalis, and none was positive for C. parvum/hominis. Whereas, modified Ziehl-Neelsen staining allowed the detection of Cryptosporidium spp. in 3% (4/132) of examined samples. Conclusion: This survey on the presence of protozoan parasites in fresh vegetables revealed that vegetables sold in Marrakech are contaminated by these protozoan parasites, as it showed that leafy green vegetables were more susceptible for parasitic contamination than root ones. Keywords: Fresh vegetables; protozoan parasites; Marrakech; qPCR.

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Quantification and viability assays of Toxoplasma gondii in commercial “Serrano” ham samples using magnetic capture real-time qPCR and bioassay techniques

Food Microbiology ◽

10.1016/j.fm.2014.07.003 ◽

2015 ◽

Vol 46 ◽

pp. 107-113 ◽

Cited By ~ 28

Author(s):

M. Gomez-Samblas ◽

S. Vílchez ◽

J.C. Racero ◽

M.V. Fuentes ◽

A. Osuna

Keyword(s):

Toxoplasma Gondii ◽

Real Time ◽

Magnetic Capture ◽

Real Time Qpcr

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Survival of giardia, cryptosporidium, poliovirus and salmonella in marine waters

Water Science & Technology ◽

10.2166/wst.1997.0744 ◽

1997 ◽

Vol 35 (11-12) ◽

pp. 261-268 ◽

Cited By ~ 30

Author(s):

D. C. Johnson ◽

C. E. Enriquez ◽

I. L. Pepper ◽

T. L. Davis ◽

C. P. Gerba ◽

...

Keyword(s):

Water Quality ◽

Salmonella Typhimurium ◽

Cryptosporidium Parvum ◽

Common Method ◽

Waterborne Pathogens ◽

Protozoan Parasites ◽

Marine Waters ◽

Untreated Sewage ◽

Cryptosporidium Parvum Oocysts ◽

Giardia Muris

Discharge of sewage into the ocean is still a common method of disposal worldwide. Both treated and untreated sewage may contain significant concentrations of waterborne pathogens, such as Giardia, Cryptosporidium, poliovirus and Salmonella. Limited studies exist on the survival of poliovirus and Salmonella in marine waters; however, almost no information exists on the survival of protozoan parasites in marine waters. This study examined the survival of Giardia muris cysts, Cryptosporidium parvum oocysts, poliovirus-1 and Salmonella typhimurium in marine waters. The survival of the microorganisms varied according to the presence of light, salinity and water quality (as determined by quantity of enterococci). All microorganisms survived longer in the dark than in sunlight, the order of survival in sunlight being: Cryptosporidium > poliovirus > Giardia > Salmonella.

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Probe-Based Real-Time qPCR Assays for a Reliable Differentiation of Capripox Virus Species

Microorganisms ◽

10.3390/microorganisms9040765 ◽

2021 ◽

Vol 9 (4) ◽

pp. 765

Author(s):

Janika Wolff ◽

Martin Beer ◽

Bernd Hoffmann

Keyword(s):

Real Time ◽

Animal Health ◽

Diagnostic Methods ◽

Virus Species ◽

Time Saving ◽

Robust Detection ◽

Highly Sensitive ◽

Reliable Differentiation ◽

World Organization ◽

Real Time Qpcr

Outbreaks of the three capripox virus species, namely lumpy skin disease virus, sheeppox virus, and goatpox virus, severely affect animal health and both national and international economies. Therefore, the World Organization for Animal Health (OIE) classified them as notifiable diseases. Until now, discrimination of capripox virus species was possible by using different conventional PCR protocols. However, more sophisticated probe-based real-time qPCR systems addressing this issue are, to our knowledge, still missing. In the present study, we developed several duplex qPCR assays consisting of different types of fluorescence-labelled probes that are highly sensitive and show a high analytical specificity. Finally, our assays were combined with already published diagnostic methods to a diagnostic workflow that enables time-saving, reliable, and robust detection, differentiation, and characterization of capripox virus isolates.

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Molecular Characterization of Giardia duodenalis in Children and Adults Sampled in Algeria

Microorganisms ◽

10.3390/microorganisms9010054 ◽

2020 ◽

Vol 9 (1) ◽

pp. 54

Author(s):

Salem Belkessa ◽

Daniel Thomas-Lopez ◽

Karim Houali ◽

Farida Ghalmi ◽

Christen Rune Stensvold

Keyword(s):

Real Time ◽

Molecular Characterization ◽

Real Time Pcr ◽

Intestinal Parasites ◽

Giardia Duodenalis ◽

Specific Pcr ◽

Stool Samples ◽

Pcr Targeting ◽

Assemblage B

The molecular epidemiology of giardiasis in Africa remains unclear. A study was carried out across four hospitals in Algeria. A total of 119 fecal samples from 55 children, 37 adults, and 27 individuals of undetermined age, all scored positive for intestinal parasites by microscopy, and were screened by real-time PCR for Giardia. Molecular characterization of Giardia was performed by assemblage-specific PCR and PCR targeting the triose phosphate isomerase gene (tpi). Of the 119 samples, 80 (67%) were Giardia-positive by real-time PCR. For 48 moderately-highly real-time PCR-positive samples, tpi genotyping assigned 22 samples to Assemblage A and 26 to Assemblage B. Contrary to Assemblage A, Assemblage B exhibited substantial genetic diversity and allelic heterozygosity. Assemblage-specific PCR proved to be specific for discriminating Assemblage A or B but not as sensitive as tpi genotyping. We confirmed that real-time PCR is more sensitive than microscopy for detecting Giardia in stool samples and that robust amplification and sequencing of the tpi gene is feasible when moderate-to-strongly real-time PCR-positive samples are used. This study is one of the few performed in Africa providing genotyping data on Giardia infections in humans. Both assemblages A and B were commonly seen and not associated with specific sociodemographic data.

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