Combined use of real-time PCR and serological techniques for improved surveillance of chronic and acute American trypanosomiasis in dogs and their owners from an endemic rural area of Neotropical Mexico.

In this study, the prevalence of T. cruzi infection was estimated in dogs and their owners from a rural community in Mexico using serological techniques for chronic infection cases, qPCR for acute phase cases, and a combination of both techniques to detect chronic and acute infections. Eighty-nine blood samples were collected from owners and their dogs for obtaining serum and parasite DNA. Prevalence was calculated using (i) positive cases detected in a serological test (ELISA and Western blot), (ii) positive cases detected in a qPCR test, and (iii) positive cases detected by both techniques. Sensitivity, specificity, and predictive values were determined individually for serology, qPCR and for both techniques used simultaneously. The prevalence observed varied: for serology, 25.8% of the dogs and 7.9% of the owners were seropositive, while for qPCR 29.2% of the dogs and 10.1% of the owners were identified as positive. Combination of serological and molecular techniques resulted in a prevalence of 38.2% for dogs and 12.4% for their owners. The sensitivity, specificity and predictive values calculated for both techniques improved when both techniques were used simultaneously (sensitivity of 92.4% and specificity of 100% for infected dogs and sensitivity of 93.4% and specificity of 100% for infected owners). Combined use of serological tests and qPCR allowed identifying a greater number of positive cases in dogs and their owners. This strategy may help implement adequate and timely epidemiological surveillance of American trypanosomiasis in order to prevent the appearance of new cases of Trypanosoma cruzi infections in endemic zones.

Screening of Zoonotic Parasites in Playground Sandboxes of Public Parks from Subtropical Mexico.

The pathological agents Toxoplasma gondii, Ancylostoma caninum, and Toxocara canis are widely distributed zoonotic parasites with high prevalence in tropical and subtropical regions of the world. The aim of the present study was to determine the presence of DNA from these parasites in sand samples from the sand playgrounds in the southeastern region of Mexico. Samples of sand were collected from 68 playgrounds in public parks in the city of Merida, Yucatan, which is the main urban area in the southeast of Mexico. The samples were examined using nested PCR to detect the SAG1 gene from Toxoplasma gondii, and endpoint PCR for the amplification of ITS-2 and rRNA-ITS2 genes from Toxocara canis and Ancylostoma caninum, respectively. The presence of T. gondii DNA was detected in 11.8% (8/68) samples, DNA from A. caninum and T. canis was not detected. Results indicate that playgrounds from the studied sandboxes are contaminated with T. gondii oocysts and may represent a risk of infection for people in contact with the sand, especially for preschoolers.

Toxoplasma gondii in Captive Wild Felids of Mexico: Its Frequency and Capability to Eliminate Oocysts.

There is little information about Toxoplasma gondii in wild felids, even when these species have been associated with cases of toxoplasmosis in humans. In this study, samples of serum and whole blood were collected from 42 felids from 10 different species, in 4 Mexican zoos. Stool samples from 36 animals were also collected, corresponding to 82% of the felids included in the study. Stool samples were used for the search of oocysts by light field microscopy and PCR. Serum samples were analyzed by indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT). DNA samples were purified from whole blood and stool for the amplification of a fragment of the SAG1 gene of T. gondii by a nested PCR (nPCR). The seroprevalence of IgG anti-T. gondii-specific antibodies by means of the ELISA was 100% (42/42) and 52.4% (22/42) by IFAT. The titers obtained varied from 1:80 to 1:2560. DNA of T. gondii was detected in 9.5% (4/42) of the blood samples by using nPCR. No oocysts were observed in the stool samples analyzed by light field microscopy. However, the DNA of the parasite was identified in 14.3% (5/35) of the stool samples evaluated. These results indicate a high prevalence of T. gondii in the studied populations of wild felids in captivity, with evidence of parasitemia and elimination of few oocysts even in adult hosts.

Presence of Toxoplasma gondii in Drinking Water from an Endemic Region in Southern Mexico.

Toxoplasmosis can be acquired through the ingestion of contaminated drinking water with oocysts of Toxoplasma gondii, highly resistant to the routinely disinfection processes; based on chlorination commonly used in the water supply industry. The aim of this study was to determine the presence of T. gondii DNA in samples of public drinking water from an endemic region of southern Mexico. In total 74 samples of water (5 L each) were collected from the three well fields (I, II, and III) and 71 independent wells, distributing public drinking water to the city of Merida Yucatan, after passing through the chlorination process. Water samples were filtered and concentrated by a sucrose solution, then DNA was extracted and evaluated through a nested-PCR (nPCR) specific for T. gondii. Positive samples were detected in 5.4% (4/74) of the water samples. This is the first report of the presence of T. gondii DNA in public drinking water from a large city in southern Mexico, where their consumption without any postpurification treatment could pose a risk for acquiring the infection in the urban population.