Dataset Information

Increasing the digital repository of DNA barcoding sequences of sand flies (Psychodidae: Phlebotominae).

ABSTRACT: Sand fly identification is complex because it depends on the expertise of the taxonomist. The females show subtle morphological differences and the occurrence of the species complexes are usual in this taxon. Therefore, a fragment of the cytochrome c oxidase subunit I (COI) gene is used for taxon barcoding to resolve this kind of problem. This study incorporates barcode sequences, for the first time, for Evandromyia cortelezzii and Migonemyia migonei from Argentina. The nucleotide sequence divergences were estimated to generate a neighbour-joining (NJ) tree. The automatic barcode gap discovery (ABGD) approach was employed to find the barcode gaps and the operational taxonomic unit (OTU) delimitation. Other species of the subtribe were included. The frequency histogram of divergences showed a barcoding gap. The ABGD analysis identified 14 operational taxonomic units (OTUs) from 13 morphological species. Sequences of Ev. cortelezzii and Mg. migonei formed well supported clusters and were diagnosed as primary species. These sequences are useful tools for molecular identification of the sand flies of the New World.

SUBMITTER: Laurito M

PROVIDER: S-EPMC6884398 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Increasing the digital repository of DNA barcoding sequences of sand flies (Psychodidae: Phlebotominae).

Laurito Magdalena M Ontivero Iliana M IM Almirón Walter R WR

Memorias do Instituto Oswaldo Cruz 20191128

Sand fly identification is complex because it depends on the expertise of the taxonomist. The females show subtle morphological differences and the occurrence of the species complexes are usual in this taxon. Therefore, a fragment of the cytochrome c oxidase subunit I (COI) gene is used for taxon barcoding to resolve this kind of problem. This study incorporates barcode sequences, for the first time, for Evandromyia cortelezzii and Migonemyia migonei from Argentina. The nucleotide sequence diver ...[more]

PMID: 31800740

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Project description:Sand fly species are traditionally identified using morphological traits, though this method is hampered by the presence of cryptic species. DNA barcoding is a widely used tool in the case of insects of medical importance, where it is necessary to know quickly which species are present in a transmission area. Here, we assess the usefulness of mitochondrial cytochrome c oxidase subunit I (COI) DNA barcoding as a practical tool for species identification, correct assignment of isomorphic females, and to evaluate the detection of cryptic diversity that occurs in the same species. A fragment of the COI gene was used to generate 156 new barcode sequences for sand flies from different countries of the Neotropical region, mainly Colombia, which had been identified morphologically as 43 species. The sequencing of the COI gene allowed the detection of cryptic diversity within species and correctly associated isomorphic females with males identified by morphology. The maximum intraspecific genetic distances ranged from 0 to 8.32% and 0 to 8.92% using uncorrected p distances and the Kimura 2-parameter (K2P) model, respectively. The minimum interspecific distance (nearest neighbor) for each species ranged from 1.5 to 14.14% and 1.51 to 15.7% using p and K2P distances, respectively. Three species had more than 3% maximum intraspecific distance: Psychodopygus panamensis, Micropygomyia cayennensis cayennensis, and Pintomyia evansi. They also were split into at least two molecular operational taxonomic units (MOTUs) each, using different species delimitation algorithms. Regarding interspecific genetic distances, the species of the genera Nyssomyia and Trichophoromyia generated values lower than 3% (except Nyssomyia ylephiletor and Ny. trapidoi). However, the maximum intraspecific distances did not exceed these values, indicating the presence of a barcode gap despite their proximity. Also, nine sand fly species were DNA barcoded for the first time: Evandromyia georgii, Lutzomyia sherlocki, Ny. ylephiletor, Ny. yuilli pajoti, Psathyromyia punctigeniculata, Sciopemyia preclara, Trichopygomyia triramula, Trichophoromyia howardi, and Th. velezbernali. The COI DNA barcode analysis enabled the correct delimitation of several Neotropical sand fly species from South and Central America and raised questions about the presence of cryptic species for some taxa, which should be further assessed.

Project description:BackgroundThe distribution of phlebotomine sand flies is changing rapidly due to climate change. This issue has implications for the epidemiology of sand fly-borne diseases, especially sand fly-associated viruses. Few studies concerning sand fly-associated viruses have been conducted in Thailand. Therefore, this study aimed to perform a molecular survey of groups of pathogenic RNA viruses belonging to the Orbivirus, Phlebovirus, and Flavivirus genera and family Rhabdoviridae in sand fly samples collected from southern Thailand.MethodsSand flies were collected at two locations in Trang and Songkhla provinces of southern Thailand, and individual sand fly samples were processed for species identification and virus detection. The Orbivirus, Phlebovirus, and Flavivirus genera and family Rhabdoviridae molecular determination was performed by RT-PCR, and positive samples were identified by cloning and sequencing, cell culture inoculation, and phylogenetic analysis.ResultsThe results presented in this study were based on the analysis of a total of 331 female sand flies. This molecular study revealed evidence of Rhabdoviridae family virus presence in Phlebotomus papatasi (3/331, 0.9%). The findings demonstrated a new cluster of rhabdovirus that was closely related to Bactrocera dorsalis sigmavirus strain BDSV.abc5 and the lineages of insect-specific Rhabdoviridae. In addition, the Bayesian tree suggested that the common ancestor of this group was the dimarhabdovirus clade. It was assumed that the virus may have switched hosts during its evolution. However, the detection of Orbivirus, Phlebovirus, and Flavivirus genera using specific primers for RT-PCR was negative in the collected sand flies.ConclusionsThere is limited knowledge on the genetic diversity and ecology of Rhabdoviridae in Thailand. This is the first data regarding the circulation of Rhabdoviridae in Ph. papatasi from Thailand. We found a new cluster of rhabdoviruses that was close to the new B. dorsalis sigmavirus. It is possible that there is a great deal of diversity in this family yet to be discovered, and a more extensive survey for new rhabdoviruses may uncover viruses from a wide diversity of host taxa and broaden our understanding of the relationships among the Rhabdoviridae.

Project description:BackgroundLeishmaniasis is an emerging vector-borne disease that occurs in Thailand. Although Leishmania (Mundinia) parasites, the causative agents of the disease have been identified, the vectors of the disease remain unidentified. In the present study, we collected sand flies from three caves located in endemic areas of leishmaniasis, including Lampang and Chiang Rai in northern Thailand, and Songkhla in southern Thailand.MethodsFemale sand flies were identified on the basis of morphological characteristics and confirmed by cytochrome c oxidase subunit I (COI) sequencing. Sand fly DNA samples were screened for Leishmania, Trypanosoma, and Bartonella DNA by polymerase chain reaction (PCR) on the basis of the ITS1 region of the ribosomal RNA (rRNA), SSU rRNA, and gltA genes, followed by phylogenetic relationships and haplotype diversity analysis.ResultsA total of 557 sand flies were identified, comprising four genera (Sergentomyia, Phlebotomus, Grassomyia, and Idiophlebotomus) and 11 species. Molecular detection of pathogens demonstrated that Leishmania DNA was not detected. However, Trypanosoma DNA was detected in 11 samples of Phlebotomus mascomai from Lampang (7 for T. noyesi), Se. anodontis from Chiang Rai (1 each for T. noyesi and Trypanosoma sp.), and Se. khawi from Songkhla (2 for Trypanosoma sp.). Bartonella DNA was detected in 16 samples of Se. anodontis and Se. barraudi s.l. from Chiang Rai, Se. anodontis from Lampang, and Se. khawi from Songkhla. The novel Bartonella sp. detected in Thai sand flies was phylogenetically related to Bartonella sp. from bats. Genetic diversity analysis showed high haplotype diversity in both Trypanosoma parasites and Bartonella bacteria.ConclusionsThe data from the present study indicate that phlebotomine sand flies could be potential vectors of zoonotic diseases caused by Trypanosoma sp. and Bartonella sp. To our knowledge, this is the first report of the natural infection of Bartonella associated with bats in Thailand, and the presence of T. noyesi and amphibian trypanosomes. However, further investigation is required to elucidate and enhance the understanding of potential vectors and transmission dynamics of pathogens in Thailand, particularly with regard to different seasonality, habitats, and host ranges.

Dataset Information

Increasing the digital repository of DNA barcoding sequences of sand flies (Psychodidae: Phlebotominae).

Publications

Increasing the digital repository of DNA barcoding sequences of sand flies (Psychodidae: Phlebotominae).

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