Toxoplasma gondii in invasive animals on the Island of Fernando de Noronha in Brazil: Molecular characterization and mouse virulence studies of new genotypes |
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| Institution: | 1. College of Marine Science, Shandong University at Weihai, Weihai, Shandong Province 264209, PR China;2. College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Province 130118, PR China;3. Jilin Entry-Exit Inspection and Quarantine Bureau, ChangChun, Jilin Province 130000, PR China;1. College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang Province 163319, PR China;2. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China;3. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, PR China;4. College of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Jinzhou, Liaoning Province 121001, PR China;5. Changchun Key Laboratory of Transomics Biotechnologies, GenSci-Changchun, Changchun, Jilin Province 130600, PR China;6. College of Basic Medicine, Mudanjiang Medical College, Mudanjiang, Heilongjiang Province 157011, PR China |
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| Abstract: | This study aimed to genetically characterize and to determine virulence from Toxoplasma gondii samples from invasive animals in the Island of Fernando de Noronha, Brazil. Blood samples were collected from 21 tegu-lizard (Salvator merianae), 12 rock-cavies (Kerodon rupestris) and 154 black-rats (Rattus rattus) from the Island and MAT (cutoff 1:25) detected anti-T. gondii antibodies in 0% of the tegus (0/21); 58.3% of the rock-cavies (7/12) and 22.7% of rats (35/154). Tissue samples (brain, heart, liver and lung) from positive animals in MAT were collected for molecular analysis and for bioassay in Swiss Webster mice. After observation period, mice were euthanized, and serological detection and tissue cyst search in the brain were performed. The brain of positive animals for serological detection or tissue cyst search was cultured in MARC-145 cells for maintenance of the T. gondii isolate. No isolate was obtained from rock cavies. Nine isolates were obtained by bioassay of 35 seropositive black rats. DNA samples were extracted from rat tissues and from parasite isolates in cell culture, and genotyped using 10 PCR-RFLP markers. ToxoDB genotypes #78 (1) from rat tissue and #146 (4), #163 (2), #260 (2) and #291 (1) from cell culture were detected. Markers of genes ROP18 and ROP5 were analyzed and in vivo virulence test was conducted in mice. Analysis revealed two allele combinations, 3/1 and 3/3, indicating non-lethal T. gondii strains, which is supported by mouse virulence test. |
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| Keywords: | Toxoplasmosis Mn-PCR-RFLP Isolation Cell culture |
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