Dataset Information

Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests.

ABSTRACT: In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii and presence of tissue cysts. We performed magnetic-capture polymerase chain reaction (MC-PCR) to detect T. gondii DNA in 231 horse meat samples purchased in supermarkets in France and evaluated the performance and level of agreement of the modified agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA) in the meat juices. The serological tests lacked sensitivity, specificity, and agreement between them, and there was no correlation with the presence of T. gondii DNA in horse meat, raising concerns about the reliability of T. gondii seroprevalence data in horses from the literature. T. gondii DNA was detected in 43% of horse meat samples but the absence of strain isolation in mice following inoculation of more than 100 horse meat samples suggests a low distribution of cysts in skeletal muscles and a low risk of T. gondii infection associated with horse meat consumption. However, to avoid any risk of toxoplasmosis, thorough cooking of horse meat is recommended.

SUBMITTER: Aroussi A

PROVIDER: S-EPMC4374124 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests.

Aroussi Abdelkrim A Vignoles Philippe P Dalmay François F Wimel Laurence L Dardé Marie-Laure ML Mercier Aurélien A Ajzenberg Daniel D

Parasite (Paris, France) 20150325

In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii and p ...[more]

PMID: 25809058

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Project description:BackgroundFree-ranging chickens are often infected with Toxoplasma gondii and seroconvert upon infection. This indicates environmental contamination with T. gondii.MethodsHere, we established a bead-based multiplex assay (BBMA) using the Luminex technology for the detection of T. gondii infections in chickens. Recombinant biotinylated T. gondii surface antigen 1 (TgSAG1bio) bound to streptavidin-conjugated magnetic Luminex beads served as antigen. Serum antibodies were detected by a fluorophore-coupled secondary antibody. Beads of differing color codes were conjugated with anti-chicken IgY or chicken serum albumin and served for each sample as an internal positive or negative control, respectively. The assay was validated with sera from experimentally and naturally infected chickens. The results were compared to those from reference methods, including other serological tests, PCRs and bioassay in mice.ResultsIn experimentally infected chickens, the vast majority (98.5%, n = 65/66) of birds tested seropositive in the BBMA. This included all chickens positive by magnetic-capture PCR (100%, n = 45/45). Most, but not all inoculated and TgSAG1bio-BBMA-positive chickens were also positive in two previously established TgSAG1-ELISAs (TgSAG1-ELISASL, n = 61/65; or TgSAG1-ELISASH, n = 60/65), or positive in an immunofluorescence assay (IFAT, n = 64/65) and in a modified agglutination test (MAT, n = 61/65). All non-inoculated control animals (n = 28/28, 100%) tested negative. In naturally exposed chickens, the TgSAG1bio-BBMA showed a high sensitivity (98.5%; 95% confidence interval, CI: 90.7-99.9%) and specificity (100%; 95% CI: 85.0-100%) relative to a reference standard established using ELISA, IFAT and MAT. Almost all naturally exposed chickens that were positive in bioassay or by PCR tested positive in the TgSAG1bio-BBMA (93.5%; 95% CI: 77.1-98.9%), while all bioassay- or PCR-negative chickens remained negative (100%; 95% CI: 85.0-100%).ConclusionsThe TgSAG1bio-BBMA represents a suitable method for the detection of T. gondii infections in chickens with high sensitivity and specificity, which is comparable or even superior to other tests. Since assays based on this methodology allow for the simultaneous analysis of a single biological sample with respect to multiple analytes, the described assay may represent a component in future multiplex assays for broad serological monitoring of poultry and other farm animals for various pathogens.

Project description:BackgroundLittle is known about the epidemiology of Toxoplasma gondii and Neospora caninum infections in alpacas (Vicugna pacos) and llamas (Lama glama) outside South America. The study aimed to estimate the seroprevalence of T. gondii and N. caninum infections in South American camelids (SAC) in Switzerland, to optimize serological tests for SAC and to identify risk factors, which may favour infection.MethodsA total of 571 sera from 132 Swiss farms (374 alpacas and 197 llamas, mean 4.3 animals/farm) were obtained. Four commercial enzyme-linked immunosorbent assays (ELISA) for detecting antibodies against T. gondii (ID Screen® Toxoplasmosis Indirect (TOXO-MS)) or N. caninum (i.e. ID Screen® Neospora caninum Indirect Multi-species (NCS-MS); ID Screen® Neospora caninum Competition (NCC) and ID Screen® Neospora caninum Indirect (NCS)) were first assessed for their use on SAC comparing their results with those in immunoblot, and optimizing cut-offs. Subsequently, two kits (TOXO-MS and NCS-MS) were selected for seroprevalence estimation. Additionally, a risk factor analysis for infection was performed on 41 farms, which agreed to participate in a web-based survey.ResultsThree kits (TOXO-MS, NCS-MS and NCC) showed almost perfect agreement (kappa > 0.901) with immunoblot results when the cut-offs were optimized, and one kit (NCS) proved not to be useful for detecting N. caninum seropositive SAC. By TOXO-MS ELISA, 82.3% (308/374) of the alpacas and 84.8% (167/197) of the llamas were seropositive for T. gondii, and 131/132 (99.2%) farms had seropositive animals. By NCS-MS ELISA, 3.5% (13/374) of the alpacas and 2.5% (5/197) of the llamas evidenced antibodies against N. caninum, and 9.1% (12/132) of the farms had seropositive animals. The variables "age" and "female sex" were identified as risk factors for T. gondii infection and "absence of cats in the farm during the last two years" as a protective factor. No risk or protective factors for N. caninum infection could be identified.ConclusionsThis nationwide cross-sectional study demonstrated for the first time the presence of antibodies against T. gondii and N. caninum in the Swiss SAC population, highlighting a high seroprevalence for T. gondii, the presence of cats as a risk factor and suggesting that SAC meat might represent an additional infection source for humans.

Project description:BackgroundThe consumption of uncooked or undercooked food from infected intermediate hosts can result in Toxoplasma gondii infection in humans. However, few studies have investigated the genetic diversity of this protozoan parasite in Iran. The aim of the present study was to genetically characterize isolates of T. gondii from intermediate host animals in Mazandaran Province, Iran.MethodsBlood and heart tissue samples were collected from 204 ruminants, and brain tissue was collected from 335 birds. The prevalence of T. gondii infection in these samples was determined serologically using the modified agglutination test and by conventional PCR assays. Those PCR samples positive for T. gondii DNA and 13 DNA samples extracted from aborted fetuses in a previous study were genotyped with 12 genetic markers using the multilocus-nested PCR-restriction fragment length polymorphism (Mn-PCR-RFLP) technique.ResultsAntibodies for parasites were found in 35.7% of the ruminant (39.1% of sheep and 26.4% of goats) samples and in 51.3% of the bird (100% of geese, 52.3% of free-range chickens and 46% of ducks) samples. Molecular detection by PCR of the repetitive 529-bp DNA fragment revealed contamination of 13.2% of ruminant (14.6% of sheep and 9.4% of goats) samples and of 9.6% of bird (11.1% of chickens, 5.7% of ducks and 0% of geese samples). The results from 30 DNA samples revealed five ToxoDB (genome database for the genus Toxoplasma) PCR-RFLP genotypes, including #1 (Type II), #2 (Type III), #10 (Type I), #27 and #48, with genotype #1 the most predominant.ConclusionsAs evidenced by the results of this study, ruminants and birds are infected with T. gondii in the region, suggesting that they are likely to be involved in the transmission of T. gondii to humans through meat consumption. The identification of different genotypes may suggest a higher genetic diversity of this parasite in Mazandaran, reflecting local environmental contamination. These results have important public health implications for the prevention and control strategies of infection.

Dataset Information

Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests.

Publications

Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests.

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