Recent studies carried out in Brazil have shown that strains from the same Toxoplasma gondii genotype can infect humans, domestic animals (dogs and cats) and animals slaughtered for human consumption (pigs, sheep, goats, and chickens), suggesting a common infection route. However, little is known about the importance of free-living wild birds within this epidemiological context. The objective of this work was to isolate, genotype, and evaluate the virulence for mice of new isolates of T. gondii obtained from free-living wild birds from the state of Minas Gerais, Southeastern Brazil. From August 2016 to June 2017, T. gondii was isolated from the hearts and brains collected from 6 out of 45 free-living wild birds, namely, a roadside hawk (Rupornis magnirostris), a campo flicker (Colaptes campestris), a southern caracara (Caracara plancus) and a tropical screech-owl (Megascops choliba), all rescued in Belo Horizonte. One isolate was obtained from a toco toucan (Ramphastos toco), rescued in Cristiano Otoni, and another was obtained from southern caracara, rescued in Santa Luzia. Five different genotypes were identified by PCR-RFLP. A unique genotype was shared in two different isolates obtained from a southern caracara and a toco toucan. This genotype has never been previously described in any other host or place. Three isolates were classified as of intermediary virulence and three isolates as avirulent for mice. The combined analysis of alleles ROP18/ROP5 (a serine/threonine kinase, and a polymorphic pseudokinase, respectively) was effective in determining the virulence of five of all the isolates with the exception of that from R. magnirostris. Atypical isolates of T. gondii obtained from free-living wild birds rescued in the state of Minas Gerais share the same genotypes of strains that infect humans, domestic animals, and animals slaughtered for human consumption.
Toxoplasma gondii presents a high prevalence worldwide, infecting several animals. Felines are considered the definitive hosts and among the intermediate hosts we highlight mammals and birds. The man can become infected by ingesting tissue cysts present in birds and mammals. Biological and molecular aspects of T. gondii allows a better understanding of the epidemiology of toxoplasmosis. This work is a serologic screening of 58 chickens grown (Gallus gallus domesticus) for human consumption in Espírito Santo State, by means of indirect haemagglutination assay (IHA). Thirteen chickens tested positive for anti-T. gondii antibodies. The heart and brain of five positive chickens were harvested, treated with pepsin and inoculated separately, in two Swiss mice, intraperitoneally. Tachyzoites were observed in the peritoneum of all the animals, between seven and 10 days after the inoculum. Ten isolates were obtained and biologically characterised in BALB/c mice inoculated with 101 to 104 tachyzoites. All isolates were classified as virulent or intermediately virulent. Isolates were genotyped by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, revealing three different genotypes. None of the isolates exhibited the clonal type I, II or III genotype. No genotypic differences were observed between the isolates from the brain or heart from the same bird.
This study aimed to identify members of the Sarcocystidae family in naturally infected wild birds at a rescue center in the state of Minas Gerais, southeastern Brazil. The heart and brain of 44 wild birds were evaluated by bioassay in mice to detect T. gondii, and extracted DNA was used for nested PCR of the 18S ribosomal DNA gene to detect members of the Sarcocystidae family. The positive samples were sequenced, assembled, edited and compared with sequences deposited in GenBank. Toxoplasma gondii was isolated from six (13.6%) out of 44 birds. Toxoplasma gondii DNA was identified in 10/44 (22.7%) of the birds. The amplified sequences exhibited 100% similarity with the DNA of the ME49 strain of T. gondii. Sarcocystis DNA (99% similarity) was identified in 5/44 (11.4%) of the birds. T. gondii and Sarcocystis spp. are common in wild birds in Minas Gerais, Brazil.
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