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Identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by righ-resolution melting analysis.

ABSTRACT: The objective of this study was to standardize the high-resolution melting method for identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by amplification of 18S ribosomal DNA (rDNA) using a single primer pair. The analyses were performed on individual reactions (containing DNA from a single species of a protozoan), on duplex reactions (containing DNA from two species of protozoa in each reaction), and on a multiplex reaction (containing DNA of four parasites in a single reaction). The proposed method allowed us to identify and discriminate the four species by analyzing the derivative, normalized, and difference melting curves, with high reproducibility among and within the experiments, as demonstrated by low coefficients of variation (less than 2.2% and 2.0%, respectively). This is the first study where this method is used for discrimination of these four species of protozoa in a single reaction.

SUBMITTER: Fehlberg HF

PROVIDER: S-EPMC5367704 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by righ-resolution melting analysis.

Fehlberg Hllytchaikra Ferraz HF Maciel Bianca Mendes BM Albuquerque George Rêgo GR

PloS one 20170327 3

The objective of this study was to standardize the high-resolution melting method for identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by amplification of 18S ribosomal DNA (rDNA) using a single primer pair. The analyses were performed on individual reactions (containing DNA from a single species of a protozoan), on duplex reactions (containing DNA from two species of protozoa in each reaction), and on a multiplex reaction (contain ...[more]

PMID: 28346485

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Project description:Simple Summary Neospora caninum, Toxoplasma gondii, Cryptosporidium and Eimeria species are parasites of phylum Apicomplexa, which includes several protozoa affecting animals and humans. In Ecuador, the maintenance of goat health is a matter of utmost importance because it affects the economic welfare of the breeders. N. caninum and T. gondii cause reproductive problems in goats, leading to abortions or weak offspring. Severe diarrhea in kids and delays in growth are due to the Cryptosporidium and Eimeria species. Moreover, T. gondii and Cryptosporidium are zoonotic parasites with serious consequences for human health. The aim of this work was to determine, by serological and parasitological tests, the prevalence of these parasites and the risk factors for goat populations in Southern Ecuador. On some farms, the prevalence of N. caninum and T. gondii reached more than 50%; up to 17% of the kids were positive for Cryptosporidium and 90% of the goats were positive for the Eimeria species. The analysis of risk factors revealed differences according to the parasite species. Considering the zoonotic significance of these results, control and prevention measures are essential and constitute a warning to veterinarians and governmental institutions. Abstract Neospora caninum, Toxoplasma gondii, Cryptosporidium and Eimeria cause severe impacts on the productivity of goat herds. The objectives of the present study were to establish the prevalence of these apicomplexans in goat farms from Ecuador; to evaluate a rapid test for Cryptosporidium diagnosis and to identify the risk factors associated with the infections. A questionnaire was designed to obtain information from 24 goat farms from Zapotillo, Garza Real, Cazaderos, Limones and Paletillas parishes in Ecuador. Blood (n = 388) and feces (n = 391) samples were collected. Indirect ELISA and standard parasitological assays were carried out to evaluate the seroprevalence of N. caninum and T. gondii and to detect oocysts of Cryptosporidium and Eimeria. The overall prevalence values of N. caninum and T. gondii were 12.11% and 18.20%, Cryptosporidium spp. and Eimeria spp. oocysts were detected in 10.49% and 89.51% of the total samples. A low correlation value was found between the results obtained by Ziehl-Nielsen and the rapid test. The multinomial logistic regression analysis revealed that vitamin supplementation, age of diarrhea, frequency of deworming, pasture area, presence of artiodactyls, domestic fowl, administration of sulfas, age group, body condition, abortions, type of pastures and the presence of cattle were risk factors according to the parasite species.

Dataset Information

Identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by righ-resolution melting analysis.

Publications

Identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by righ-resolution melting analysis.

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