Table 1.
Details of the molecular techniques for DNA amplification used in the present study.
Fig 1.
Representative amplicons of PCR assays in blood samples of wild animals visualized on 2% agarose gel after electrophoresis and ethidium bromide staining.
[A]. Mammalian IRBP gene amplification by conventional PCR (cPCR). Sample numbers are indicated on top. Positive amplification is indicated by the presence of a 227-bp band (indicated by the arrow). PC – Positive control (DNA isolated from dog skin); NC – Negative control (no DNA); MM – 120-bp molecular marker. [B]. DNA amplification with primers for the SSU rRNA gene (round 2 of LnPCR) of Leishmania sp. Samples are identified on top. Positive amplification is indicated by the presence of a 358-bp band (indicated by the arrow). PC - positive control (DNA from Leishmania infantum reference strain MHOM/BR/PP75); NC - negative control (no DNA); MM - 120 bp molecular marker. The positive sample for Leishmania DNA (B19 from maned wolf) is boxed in [A] and [B]. Photo credits: NLCP.
Table 2.
Molecular detection of Leishmania spp. DNA in skin and blood samples from wild mammals at the Belo Horizonte Zoo, Minas Gerais, Brazil (April 2022-October 2023).
Fig 2.
Visual readout of K26-LAMP assay for Leishmania DNA detection.
PC - Positive control (DNA from Leishmania infantum reference strain MHOM/BR/PP75); NC - Negative control (no DNA); B19 – Positive blood sample from Chrysocyon brachyurus. Photo credits: NLCP.
Fig 3.
Maximum Likelihood phylogenetic tree inferred from SSU rRNA gene sequence of Leishmania spp.
The tree was constructed using the Juke-Cantor substitution model with 1,000 bootstrap replications. Bootstrap values ≥ 70% are shown at the nodes. The sequence obtained from the maned wolf (Chrysocyon brachyurus) in this study (GenBank deposit PV 977395) clusters within the Leishmania infantum clade (highlighted in bold). Reference sequences from GenBank are labeled with their accession numbers followed by species names. Trypanosoma cruzi (GenBank accession JF746736.1) was used as the outgroup.