Molecular Detection and Genetic Diversity of Toxoplasma gondii Oocysts in Cat Faeces from Klang Valley, Malaysia, Using B1 and REP Genes in 2018

Abstract

The major route for Toxoplasma gondii (T. gondii) infection is through the ingestion of foods contaminated with oocyst from cat faeces. The microscopic detection of T. gondii oocysts in cat faeces is challenging, which contributes to the failure of detecting or differentiating it from other related coccidian parasites. This study aims to detect T. gondii oocysts in cat faeces using two multicopy-target PCR assays and to evaluate their genetic diversity. Cat faecal (200) samples were collected from pet cats (PCs; 100) and free-roaming cats (FRCs; 100) within Klang Valley, Malaysia, and screened for coccidian oocysts by microscopy using Sheather’s sucrose floatation. PCR assays were performed on each faecal sample, targeting a B1 gene and a repetitive element (REP) gene to confirm T. gondii oocysts. Additionally, the PCR amplicons from the REP gene were sequenced to further confirm T. gondii-positive samples for phylogenetic analysis. Microscopy detected 7/200 (3.5%) T. gondii-like oocysts, while both the B1 gene and the REP gene detected 17/200 (8.5%) samples positive for T. gondii. All samples that were microscopically positive for T. gondii-like oocysts were also shown to be positive by both B1 and REP genes. The BLAST results sequenced for 16/200 (8.0%) PCR-positive T. gondii samples revealed homology and genetic heterogeneity with T. gondii strains in the GenBank, except for only one positive sample that did not show a result. There was almost perfect agreement (k = 0.145) between the two PCR assays targeting the B1 gene and the REP gene. This is the first report on microscopic, molecular detection and genetic diversity of T. gondii from cat faecal samples in Malaysia. In addition, the sensitivities of either the B1 gene or REP gene multicopy-target PCR assays are suitable for the accurate detection of T. gondii from cat faeces.

Prevalence and Molecular Characterization of Toxoplasma gondii and Toxocara cati Among Stray and Household Cats and Cat Owners in Tehran, Iran

“…For example, the prevalence of T. gondii in stray cats in Tehran (15.2%), compared to other molecular studies in Iran, was higher than the previous prevalence reported in Mashhad (4.5%) ( 25 ) and Ahvaz (7.2%)( 26 ), and lower than the prevalence (24.1%) reported in Shiraz ( 27 ). Furthermore, compared to other molecular studies in the world, it is higher than the previous prevalence reported in South Korea (4.7% and 1.3%) ( 28 , 29 ), the USA (2%) ( 30 ), and Malaysia (13%)( 31 ), and lower than the prevalence reported in South Korea (17.5% and 30.6%)( 32 , 33 ) and Pakistan (34%)( 34 ). In addition, the prevalence of T. gondii infection in household cats in the current study (18.2%) was higher than the prevalence reported in Switzerland (0.58%) ( 35 ), Poland (2.4%) ( 36 ), Malaysia (4%) ( 31 ), Thailand (4.7%) ( 37 ) and Kenya (7.8%) ( 38 ), and lower than reported from Italy (20.5%) ( 39 ).…”

“…Furthermore, compared to other molecular studies in the world, it is higher than the previous prevalence reported in South Korea (4.7% and 1.3%) ( 28 , 29 ), the USA (2%) ( 30 ), and Malaysia (13%)( 31 ), and lower than the prevalence reported in South Korea (17.5% and 30.6%)( 32 , 33 ) and Pakistan (34%)( 34 ). In addition, the prevalence of T. gondii infection in household cats in the current study (18.2%) was higher than the prevalence reported in Switzerland (0.58%) ( 35 ), Poland (2.4%) ( 36 ), Malaysia (4%) ( 31 ), Thailand (4.7%) ( 37 ) and Kenya (7.8%) ( 38 ), and lower than reported from Italy (20.5%) ( 39 ). To the best of our knowledge, this is the first study performed on the molecular prevalence of T. gondii infection in cat owners worldwide.…”

Development of Toxoplasma gondii Chinese I genotype Wh6 Strain in Cat Intestinal Epithelial Cells

“…All cat feces were collected daily and the production of oocysts was monitored by microscopic examination and PCR using T. gondii-specific target genes (529 bp repetitive sequence and B1 gene). Oocyst DNA was extracted from cat feces using the QIAamp Fast DNA Stool Mini Kit (Qiagen, Hilden, Germany) as previously described [21]. Two sets of T. gondii specific primers (Supplementary Table S1) were used in separate PCR reactions [22].…”

Review on diagnosis and molecular characterization of Toxoplasma gondii in humans and animals

“…Toxoplasma gondii is an important obligate intracellular apicomplexan zoonotic protozoa that causes toxoplasmosis in humans and other warm blooded animals worldwide (Wana et al, 2020a;Matta et al, 2021). Like other protozoan species, T. gondii has three infectious stages including oocysts, bradyzoites and tachyzoites (Waldman et al, 2020).…”