Larissa Pereira Rodrigues

Purpose: This study investigated the occurrence of vector-borne pathogens in free-living opossums ( Didelphis albiventris and Didelphis aurita ) from São Paulo State, Brazil, with emphasis on Babesia spp., Hepatozoon spp., and Ehrlichia spp. Methods: Blood samples from 115 opossums (95 D. albiventris and 20 D. aurita ) were analyzed using parasitological and molecular approaches. Conventional PCR assays were performed for Babesia spp. and Hepatozoon spp., whereas nested PCR was used for Ehrlichia spp. Positive amplicons were sequenced and subjected to phylogenetic analysis. Results: Five samples (4.35%) from D. albiventris collected in Botucatu were positive for Babesia spp. based on the 18 S rRNA gene, and two samples (1.74%) were positive for Ehrlichia spp. based on the dsb gene. No Hepatozoon spp. DNA was detected. Sequence analysis revealed 100% identity between the Babesia amplicons and Babesia sp. (GenBank: MW290046), while one Ehrlichia sequence showed 100% identity with Ehrlichia sp. strain Natal (GenBank: KY207546; OP270482), and the second sequence showed 99.71% identity with Ehrlichia sp. (GenBank: OP270482). Phylogenetic reconstruction clustered the Babesia sequences within the South American Marsupialia Group and positioned the Ehrlichia sequences together with lineages previously reported in South American marsupials. Conclusion: These findings provide the first molecular detection of Babesia sp. and Ehrlichia sp. in free-living D. albiventris from Botucatu, São Paulo State, Brazil. The results expand current knowledge on vector-borne pathogens in synanthropic marsupials and reinforce the importance of continued surveillance to improve understanding of host–pathogen interactions at urban–wildlife interfaces within a One Health perspective.

Mammalian hair is a source of biological information and can be used in genetic, toxicological, hormonal, and ecological studies. However, non-invasive collection methods are still little explored. This study aimed to describe and validate a passive methodology for collecting hair from jaguars (Panthera onca) and evaluate its viability for different analyses. This study was conducted in the Northern Pantanal, where synthetic fiber mats were installed in strategic locations to passively capture hair. The presence of animals and the collection of samples were monitored by camera traps over a period of 30 days. The collected samples were subjected to morphological analyses by electron microscopy, molecular tests for genetic and sex identification, and isotopic and heavy metal analyses. The results showed that the collected hairs were well preserved, allowing the structural and molecular identification of the material. The analyses confirmed the viability of DNA for genetic studies and revealed specific concentrations of heavy metals and stable isotopes. The proposed methodology proved to be effective and is a promising alternative for obtaining samples without directly interfering with the behavior of the animals.