Dataset Information

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

ACCESS DATA

Publications

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

Similar Datasets

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.

Project description:BACKGROUND:Knowledge about parasitic infections is crucial information for animal health, particularly of free-ranging species that might come into contact with livestock and humans. METHODS:We investigated the seroprevalence of three tissue-cyst-forming apicomplexan parasites (Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti) in 506 individuals of 12 wildlife species in Namibia using in-house enzyme linked immunosorbent assays (indirect ELISAs applying purified antigens) for screening and immunoblots as confirmatory tests. We included six species of the suborder Feliformia, four species of the suborder Caniformia and two species of the suborder Ruminantia. For the two species for which we had most samples and life-history information, i.e. cheetahs (Acinonyx jubatus, n?=?250) and leopards (Panthera pardus, n?=?58), we investigated T. gondii seroprevalence in relation to age class, sex, sociality (solitary, mother-offspring group, independent sibling group, coalition group) and site (natural habitat vs farmland). RESULTS:All but one carnivore species (bat-eared fox Otocyon megalotis, n?=?4) were seropositive to T. gondii, with a seroprevalence ranging from 52.4% (131/250) in cheetahs to 93.2% (55/59) in African lions (Panthera leo). We also detected antibodies to T. gondii in 10.0% (2/20) of blue wildebeest (Connochaetes taurinus). Adult cheetahs and leopards were more likely to be seropositive to T. gondii than subadult conspecifics, whereas seroprevalence did not vary with sex, sociality and site. Furthermore, we measured antibodies to N. caninum in 15.4% (2/13) of brown hyenas (Hyaena brunnea) and 2.6% (1/39) of black-backed jackals (Canis mesomelas). Antibodies to B. besnoiti were detected in 3.4% (2/59) of African lions and 20.0% (4/20) of blue wildebeest. CONCLUSIONS:Our results demonstrate that Namibian wildlife species were exposed to apicomplexan parasites at different prevalences, depending on parasite and host species. In addition to serological work, molecular work is also needed to better understand the sylvatic cycle and the clear role of wildlife in the epidemiology of these parasites in southern Africa.

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.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets