Molecular detection and phylogenetic characterization of Babesia and Ehrlichia in free-living opossums from southeastern Brazil

Abstract

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.

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