Molecular investigation on the occurrence of Toxoplasma gondii oocysts in cat feces using TOX-element and ITS-1 region targets

Chemoh, Waenurama; Sawangjaroen, Nongyao; Nissapatorn, Veeranoot; Sermwittayawong, Natthawan

doi:10.1016/j.tvjl.2016.05.018

Cited by 30 publications

(37 citation statements)

References 38 publications

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“…This result may be due to consumption of meat contaminated with Toxoplasma cysts or due to the small number of Toxoplasma oocysts in fecal specimens which are difficult to determine by fecal examination. Similar to our results, a low prevalence of oocysts shedding in cats was determined at 1.2% in Iran [16], at 2.3% in Italy [17], at 0.3% in Japan [18], at 0.14 % in Germany [19], at 0.4% in Switzerland [20], at 4.7% in South Korea [21], at 0.9 in the USA [22], at 0.8% in Thailand [23], at 0.76 % in Finland [24] by microscopy and PCR methods. In contrast to serological studies, examining feces did not show any information about age and gender as risk factors, due to the low prevalence of oocyst shedding by the cats [13,25].…”

Section: Discussionsupporting

confidence: 90%

“…These five markers have been successfully used in T. gondii genotypes in cats in previous studies and proved to identify the genotype of isolates in cats [29][30][31]. Although, ten genetic markers allow isolates with high resolution [32]. In Iran, types II and III isolates have been detected by RLFP analysis at GRA6 locus [33,34] and type I and III by RLFP analysis at SAG 2 locus in stray cats [35].…”

Section: Discussionmentioning

confidence: 99%

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Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran.

Khodaverdi

Razmi

2019

Preprint

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Background: Cats as a definitive host have an important role in the epidemiology of toxoplasmosis in humans and animals. The aim of the study was to determine the frequency of Toxoplasma gondii infection and isolate and identify the genotypes of T. gondii in stray cats in Mashhad suburb. Methods: From April 2016 to August 2017, 175 fecal samples from stray cats and 31 brain samples from cats killed in driving accidents were collected. The fecal samples were examined by fecal flotation technique and T. gondii -specific PCR. The brain samples were investigated by T. gondii -specific PCR and consequently examined by mice bioassay. The DNA of T. gondii isolated was genotyped using SAG1, SAG2, SAG3, BTUB and GRA6 as PCR-restriction fragment length polymorphism (PCR-RFLP) markers. Results: In the present study, Toxoplasma -like oocysts were microscopically observed in 2.2% (4/175) fecal samples. The presence of Toxoplasma oocysts was confirmed in one microscopy-positive sample by PCR. In addition, T . gondii DNA was detected in 4% (7/175) microscopy-negative samples using PCR. T. gondii was isolated from one brain PCR-positive sample by mice bioassay. The isolate was avirulent and many T. gondii cysts were observed in mice brain. The isolate was successfully genotyped by PCR-RLFP analysis .The isolated genotyped was type II. Beside, eight Toxoplasma- positive fecal samples contained insufficient DNA and only amplified at SAG-3 locus in PCR. These samples were also showed type II pattern at this locus. Conclusions: Parasitological and molecular results showed low frequency of Toxoplasma infection in the stray cats, and identified the genotype of T. gondii isolate as type II, for the first time in Mashhad area, Khorasan Razavi Province.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran.

Khodaverdi

Razmi

2019

Preprint

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Section: Discussionsupporting

confidence: 91%

“…However, a single marker was used to genotype Toxoplasma in these studies but it does not allow identification of nonclonal strains, and to determine more precisely the presence of polymorphisms in the population [7,38]. Our result also agreed with similar studies in other countries that type II was detected in stray cats using PCR-RLFP [22,25,31,[39][40][41][42][43].…”

Section: Discussionsupporting

confidence: 85%

“…The PCR-RFLP assay at five markers revealed that the avirulent isolate in this study belongs to type II clonal lineage. These five markers have been successfully used in T. gondii genotypes in cats in previous studies and proved to identify the genotype of isolates in cats [29][30][31]. Although, ten genetic markers allow isolates with high resolution [32].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran.

Khodaverdi

Razmi

2019

Preprint

View full text Add to dashboard Cite

Background: Cats as a definitive host have an important role in the epidemiology of toxoplasmosis in humans and animals. The aim of the study was to determine the frequency of Toxoplasma gondii infection and isolate and identify the genotypes of T. gondii in stray cats in the Mashhad suburb. Methods: From April 2016 to August 2017, 175 fecal samples from stray cats and 31 brain samples from cats killed in driving accidents were collected. The fecal samples were examined by fecal flotation technique and T. gondii-specific PCR. The brain samples were investigated by T. gondii-specific PCR and consequently examined by mice bioassay. The DNA of T. gondii isolated was genotyped using SAG1, SAG2, SAG3, BTUB and GRA6 as PCR-restriction fragment length polymorphism (PCR-RFLP) markers. Results: In the present study, Toxoplasma-like oocysts were microscopically observed in 2.2% (4/175) fecal samples. The presence of Toxoplasma oocysts was confirmed in one microscopy-positive sample by PCR. In addition, T. gondii DNA was detected in 4% (7/175) microscopy-negative samples using PCR. T. gondii was isolated from one brain PCR-positive sample by mice bioassay. The isolate was avirulent and many T. gondii cysts were observed in mice brain. The isolate was successfully genotyped by PCR-RLFP analysis .The isolated genotyped was type II. Besides, eight Toxoplasma-positive fecal samples contained insufficient DNA and only amplified at SAG-3 locus in PCR. These samples were also showed type II pattern at this locus. Conclusions: Parasitological and molecular results showed low frequency of Toxoplasma infection in the stray cats, and identified the genotype of T. gondii isolate as type II, for the first time in Mashhad area, Khorasan Razavi Province.

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Detection of Toxoplasma gondii in faeces of privately owned cats using two PCR assays targeting the B1 gene and the 529-bp repetitive element

et al. 2017

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Toxoplasma gondii infection is a worldwide parasitic zoonosis with a high-health risk for humans. The key epidemiological role played by felids is related to oocyst shedding. The present study compared two amplification protocols for the molecular diagnosis of Toxoplasma infection in owned cats. A total of 78 owned cats referred to an Italian university-teaching hospital and exposed to various T. gondii-associated risk factors were sampled for blood and faeces. Faecal specimens were processed by flotation and tested using 2 copro-PCRs targeting the widely used B1 gene and the 529-bp repetitive element (RE). The sera were tested by the indirect immunofluorescent antibody test (IFAT) for the detection of immunoglobulins against T. gondii. Sixteen faeces (20.52%) tested positive for T. gondii DNA; 12 samples were positive only at B1-PCR, two at 529-bp RE-PCR and two at both genetic targets (overall agreement = 82.11%). The amplicons obtained were sequenced, and the Basic Local Alignment Search Tool analysis showed a high homology with the T. gondii strains available in reference databases. Two stool samples were microscopically positive for T. gondii-like oocysts and also tested positive by both B1 and 529-bp RE-PCRs. Thirty-three (42.3%) sera tested positive for antibodies; of which, seven were found to have T. gondii DNA-positive results using the B1 genetic target (overall agreement = 57.77%). The amplification sets targeting B1 and 529-bp RE showed substantially different yields. Further research is needed to better understand the significance and the sensitivities of using these multi-copy-targeted molecular methods from cat faeces before being used for routine diagnosis.

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Molecular investigation on the occurrence of Toxoplasma gondii oocysts in cat feces using TOX-element and ITS-1 region targets

Cited by 30 publications

References 38 publications

Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran.

Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran.

Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran.

Detection of Toxoplasma gondii in faeces of privately owned cats using two PCR assays targeting the B1 gene and the 529-bp repetitive element

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