Fig 1.
The figure presents the photographic registry of dogs with lesions.
A: dog with the mucous lesion on the nose. B: dog with the scrotal lesion. C: dog presenting ulcerative lesions on the edge of the ear.
Table 1.
Detection of Leishmania in biopsy lesion samples obtained from domestic dogs by the PCR and sequence analysis by SSU rRNA and ITS1, RFLP hsp70C and HRM analysis targeting hsp70.
Fig 2.
The figure depicts the alignment of nucleotide sequences based on the SSU rRNA gene of isolates which were characterized in this study (in bold), as well as compared to sequences of Leishmania species deposited in the GenBank such as L. infantum (XR001203206), L. chagasi (KJ697713), L. amazonensis 1 (JX030083), L. amazonensis 2 (JX030084), L. amazonensis 3 (JX030085) L. guyanensis (KF041803) and L. braziliensis (JX030135).
Fig 3.
The figure shows the network genealogy using ITS1 rDNA sequences from isolates of Leishmania braziliensis (type 1 and 2) and other Leishmania spp. Numbers in nodes correspond to support values estimated by 100 bootstrap replicates using the same parameter, which was optimized for network inferences.
The isolates characterized in this study are indicated in bold and compared to other isolates of Leishmania spp. used in Van der Auwera et al. (2014) and Avila et al. (2018).
Table 2.
The table presents the hsp70 amplicons 1, 2, and 3 of DNA, from each sample, submitted to HRM analysis.
Fig 4.
The figure presents the melting temperatures (Tm) for Amplicons 1 (Fig 4A), 2 (Fig 4B), 3 (Fig 4C), and standard species.
The plots show the average Tm values. Each species and sample was tested in duplicate. INF: L. (L.) infantum; AMA: L. (L.) amazonensis; MEX: L. (L.) mexicana; LAI: L. (V.) lainsoni; BRA: L. (V.) braziliensis; GUY: L. (V.) guyanensis; NAI: L. (V.) naiffi; SHA: L. (V.) shawi. (1) Dog 01 –Chorinho; (2) Dog 02 –Hulk; (3) Dog 03 –Halley; (4) Dog 05 –Help; (5) Dog 06 –Negão; (6) Dog 11 –Valente; (7) Dog 15 –LN–Xapuri; (8) Dog 15 –LP–Xapuri; (9) Dog 26 –LN–Marmaduque; (10) Dog 26 –LP–Marmaduque; (11) Dog 37 –Xorinho 1; (12) Dog 39 –Pantera; (13) Dog 40 –Bethoven; (14) Dog 41 –Xorinho 3.
Fig 5.
The figure displays the hsp70C nested PCR and hsp70C-RFLP profiles.
PCR products (A) and RFLP products (B,) were separated in a 3% agarose gel electrophoresis, stained with ethidium bromide and visualised under UV light. INF: L. (L.) infantum; AMA: L. (L.) amazonensis; MEX: L. (L.) mexicana; LAI: L. (V.) lainsoni; BRA: L. (V.) braziliensis; GUY: L. (V.) guyanensis; NAI: L. (V.) naiffi; SHA: L. (V.) shawi. ND undigested fragment; NC: negative control (NTC from preamplification reaction used as template in the nested PCR); NTC: Non template control. (1) Dog 01 –Chorinho; (2) Dog 02 –Hulk; (3) Dog 03 –Halley; (4) Dog 05 –Help; (5) Dog 06 –Negão; (6) Dog 11 –Valente; (7) Dog 15 –LN–Xapuri; (8) Dog 15 –LP–Xapuri; (9) Dog 26 –LN–Marmaduque; (10) Dog 26 –LP–Marmaduque; (11) Dog 37 –Xorinho 1; (12) Dog 39 –Pantera; (13) Dog 40 –Bethoven; (14) Dog 41 –Xorinho 3.