Leishmania

Detection of Leishmania View in CoL and Trypanosoma DNA in sand flies

A total of 557 female sand flies were tested for

Leishmania and Trypanosoma DNA on the basis of ITS1 and SSU rRNA amplifications, respectively. Te present study demonstrates that the TRY927F and TRY927R primer sets, targeted for the SSU rRNA gene, were able to successfully amplify all positive Trypanosoma DNA. Of these, 11 (1.97%) samples for Trypanosoma DNA , 7 Trypanosoma positives belonged to Ph. mascomai from Lampang, and 2 Trypanosoma positives were detected in Se. anodontis from Chiang Rai ( Table 1). BLASTn analysis demonstrated a Trypanosoma SSU rRNA sequence length of approximately 937 bp in eight samples (PT82-10, PT84-62, PT96-49, PT132-17, PT144-69, PT154-73, PT158-28, and CR110- 13), showing the ranged 99.89–100% sequence similarities with T. noyesi (accession number OP022194) that is available in the GenBank database. Furthermore, sample CR102-12 was consistent with the 930 bp length of Trypanosoma SSU rRNA sequence, the BLASTn result demonstrated a 99.89% similarity with Trypanosoma sp. (accession number MH989552), which was detected in sand flies from southern Tailand. From the detection of Trypanosoma in Songkhla, two samples (SK4- 23 and SD33-33) were positively detected in Se. khawi . Te result demonstrated that the SSU rRNA sequences were approximately 974 bp and 931 bp in length, sharing 99.74% (accession number MH989543) and 100% (accession number MH989552) identities for SK4-23 and SD33-33, with unidentified Trypanosoma found in previously reported sand flies in the Songkhla province of southern Tailand.

Te maximum likelihood tree demonstrated that eight positive Trypanosoma were genetically classified to T. noyesi of the T. cruzi clade, which was detected in the sand flies from Tailand, as previously recorded in the GenBank database. Moreover, two positive Trypanosoma sp. (CR102-12 from Chiang Rai and SD33-33 from Songkhla) were distinctively classified into the An01 + An02/ Frog2 lineage, while a positive Trypanosoma in Songkhla (SK4-23) was genetically clustered into the An04/ Frog1 lineage of the anuran clades (Supplementary 3: Fig. S2 View Fig ). Using ITS1 -PCR, Leishmania DNA was not detected in all sand flies tested in the present study. Te Trypanosoma SSU rRNA sequences were assigned to the GenBank database with following accession numbers: OP861666-OP861676.