No abstract

In the last decade, a method widely used to delimit areas of endemism is the analysis of endemicity (AE), a non-hierarchical and grid-dependent algorithm implemented through the package NDM/VNDM. Its input files are based on lists of georeferenced taxa, and any mistakes in their preparation will influence the results of the analyses. We describe here a free online automated tool for generating the input files for VNDM from simple spreadsheets.

Introduction: Leishmaniasis, a neglected infectious disease affecting humans, domestic, and wild animals, is caused by 20 from 53 Leishmania genus species and transmitted by sandflies. Leishmania genus, belonging to the Trypanosomatidae family and Kinetoplastida order, are grouped into five subgroups according to the biogeographic and evolutive history of parasites and hosts, leading to incongruences and paraphyly. The GH18 Leishmania chitinase, which is encoded by a species-specific single-copy gene, conserved in the basal groups of trypanosomatids, and absent in the genus Trypanosoma, was evaluated as a phylogenetic marker and a diagnostic target. Methods: Primers were designed to detect Leishmania in its host biological samples and obtain the chitinase sequence of species that are unavailable in public databanks. The GH18 chitinase gene and its genomic context were evaluated phylogenetically. A protocol was developed to discriminate Leishmania subgenera by adopting polymerase chain reaction (PCR) and restriction fragment length polymorphism and using in silico tools. The adopted PCR method for detecting a partial 953 bp GH18 chitinase-encoding gene represented high sensibility and specificity on DNA of isolated parasites and was used as negative controls, Trypanosoma cruzi, and DNA from Leishmania hosts. Results: Preservation of the chitinase locus in the aquatic free-living protozoan Bodosaltans disclosed a primitive common origin. Based on the comparative analysis, the amino acid sequence of GH18 trypanosomatidae chitinase demonstrated its high similarity to that of chitinase from marine prokaryotes and protozoans. Phylogenetic reconstruction based on chitinase corroborated the Supercontinent Origins Theory for Leishmania. Conclusion: The chitinase-encoding gene was effectively detected in biological samples and thus could be considered for differential molecular diagnosis among Leishmania clinical important species worldwide.

Leishmaniasis, a neglected infectious disease affecting humans, domestic and wild animals, caused by 20 from 53 Leishmania genus species, is transmitted by sandflies. Leishmania genus, belonging to Trypanosomatide Family and Kinetoplastida Order, are grouped in five subgroups according to biogeographic and evolutive history of parasites and hosts, which has led to incongruences and paraphyly. The GH18 Leishmania chitinase, encoded by a specie-specific single copy gene, conserved in basal groups of trypanosomatids, and absent in the genus Trypanosoma, was evaluated as a phylogenetic marker and a diagnostic target. Primers were designed to detect Leishmania in its host biological samples and to obtain the chitinase sequence of species not available in public databanks. The GH18 chitinase gene and its genomic context was evaluated phylogenetically. A protocol to discriminate among Leishmania subgenera by PCR and restriction fragment length polymorphism (RFLP) was developed using in silico tools. A PCR method to detect a partial 953 bp GH18 quitinase encoding gene presented high sensibility and specificity on isolated parasites DNA and using as negative controls, Trypanosoma cruzi, and DNA from Leishmania hosts. Preservation of the chitinase locus in the aquatic free-living protozoan Bodo saltans, disclose a primitive common origin. GH18 trypanosomatide chitinase amino acid sequence comparative analysis revealed high similarity to chitinase from marine prokaryotes and protozoan. Phylogenetic reconstruction based on chitinase corroborates the Supercontinent Origins theory for Leishmania. The chitinase encoding gene was effectively detected in biological samples and for differential molecular diagnosis among Leishmania clinical important species worldwide.

Leishmaniasis, an infectious disease that affects humans, domestic dogs, and wild animals, is caused by 20 of the 53 Leishmania genus species and is transmitted by sandflies. Despite its significant impact, the disease is often neglected. Leishmania genus, belong to Trypanosomatide Family and Kinetoplastida Order, are grouped in five subgroups according to biogeographic and evolution history of parasites and hosts. The GH18 Leishmania chitinase is encoded by a specie-specific single copy gene, conserved in basal groups of trypanosomatids, and is absent in the genus Trypanosoma. Preservation of the chitinase genomic locus in the aquatic free-living protozoan Bodo saltans, discloses a primitive common origin. Trypanosomatid chitinase amino acid sequence comparative analysis revealed high similarity with chitinase from sea living prokaryotes and protozoan microorganisms, indicating a probable marine origin. Amino acid sequence comparative analysis revealed that perhaps the trypanosomatid chitinase derived from a water living Kinetoplastida ancestor and its phylogenetic reconstruction corroborates the Supercontinent Origins theory for Leishmania. The chitinase-encoding gene was effective for differential molecular diagnosis among Leishmania clinical important species worldwide.