The aim of the present study was to detect DNA of Toxoplasma gondii in Crassostrea spp. oysters cultured in the state of Pará, Brazil. A total of 400 oysters were directly collected from a fixed rack system. Gills, gastrointestinal tract (GIT) and intervalvular liquid were separated and grouped into pool samples of 10 animals, resulting in 40 samples each of gills, GIT and intervalvular liquid. DNA extraction was performed using a commercial kit, and T. gondii DNA was detected by nested PCR using the primers Toxo3 and Toxo4, which produced an amplification product of 155 bp of the T. gondii gene B1. Nucleotide sequencing was performed for positive samples, and the obtained sequences were identified by comparison with sequences in GenBank. The DNA of T. gondii was detected in 5.8% (7/120) of the pool samples, of which 7.5% (3/40) was in the GIT, 5% (2/40) in the gills, and 5% (2/40) was in the intervalvular liquid. The obtained sequences presented 100% identity and overlap with T. gondii DNA sequences. This is the report of detection of T. gondii DNA in oysters from genus Crassostrea spp. originating from the state of Pará, eastern Amazon.

Leptospirosis is a zoonosis of worldwide distribution in which the agent can infect several animal species and accidentally humans. In view of the limited number of studies on anti-Leptospira antibodies in wild animal species, especially those living in aquatic environments, we sought in this study to investigate the presence of these antibodies in the spot-legged turtle (Rhinoclemmys punctularia) maintained in captivity in the Rodrigues Alves Botanical Garden-Amazon Zoobotanical Park, located in Belém, Pará State, Brazil. Serum samples were collected from 31 turtles, and identification of anti-Leptospira antibodies was performed using the microscopic agglutination test, using a collection of 31 live antigens which represent 19 serogroups of Leptospira. Among the analyzed samples, 54.83% (17/31) were observed to be reactive, and co-agglutination was detected in a further six samples which were not accounted for in the frequency of serogroups. The most frequently detected serogroups were Tarassovi 72.72% (8/11), Celledoni 18.18% (2/11), and Pomona 9.09% (1/11)], with titers ranging from 200 to 400, being this the first study to report infection of these serogroups in this species of chelonios. The animals were kept in water tanks, which probably favored the transport of the agent and allowed its transmission to the spot-legged turtle. We thus confirmed presence of anti-Leptospira antibodies in chelonians maintained in the Rodrigues Alves Botanical Garden.

We analyzed the presence of Leptospira spp. in liver and kidney tissue of wild marsupials and rodents trapped in a periurban forest in the eastern Brazilian Amazon. We examined 25 individuals of four marsupial and seven rodent species for the presence of the 16S rRNA gene of Leptospira in the DNA extracted from 47 liver and kidney tissue samples using PCR. We detected positive samples in 12% (3/25) of the individuals, in kidney fragments of two marsupial species (Didelphis marsupialis and Marmosops pinheiroi) and in a liver fragment of one rodent species (Echimys chrysurus). These are the first records of Leptospira spp. in M. pinheiroi and E. chrysurus and it is the first molecular survey of marsupials and rodents in the Brazilian Amazon.