DNA barcode for identification of immature stages of sand flies (Diptera: Psychodidae) collected from natural breeding sites

Vivero, Rafael José; Bejarano, Eduar Elías; Estrada, Luis Gregorio; Flórez, Fernando Castro; Ortega-Gómez, Edgar; Aparicio, Yamileth; Gutiérrez, Carolina Torres; Uribe, Sandra; Muskus-López, Carlos Enrique

doi:10.11646/zootaxa.4277.2.3

Cited by 15 publications

(7 citation statements)

References 30 publications

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“…cryptic species and interspecific sexual isomorphism). Thus, molecular identification methods, such as DNA barcoding, based on the amplification of a fragment of the mitochondrial cytochrome c oxidase subunit I ( coi ) gene (Hebert et al ., 2003) can be used to distinguish between isomorphic females belonging to different species (Srinivasan et al ., 2014; Nzelu et al ., 2015; Pinto et al ., 2015; Rodrigues et al ., 2018), associate immature and adult life‐cycle stages (Vivero et al ., 2017) and uncover both intra‐ and interspecific cryptic diversity within sandflies (Kumar et al ., 2012; Pinto et al ., 2015; Polseela et al ., 2016; Rodrigues et al ., 2018).…”

Section: Introductionmentioning

confidence: 99%

Single‐locus DNA barcoding and species delimitation of the sandfly subgenus Evandromyia (Aldamyia)

Rodrigues¹,

Baton²,

Shimabukuro

2020

Medical Vet Entomology

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Sandfly specimens from the subgenus Evandromyia (Aldamyia) Galati, 2003 (Diptera: Psychodidae: Phlebotominae) were collected between 2012 and 2019 from nine localities in seven Brazilian states, morphologically-identified, and then DNA barcoded by sequencing the mitochondrial cytochrome c oxidase subunit I (coi) gene. Forty-four new barcode sequences generated from 10 morphospecies were combined with 49 previously published sequences from the same subgenus and analysed using sequence-similarity methods (best-match criteria) to assess their ability at specimen identification, while four different species delimitation methods (ABGD, GMYC, PTP and TCS) were used to infer molecular operational taxonomic units (MOTUs). Overall, seven of the 11 morphospecies analysed were congruent with both the well-supported clades identified by phylogenetic analysis and the MOTUs inferred by species delimitation, while the remaining four morphospecies-E. carmelinoi, E. evandroi, E. lenti and E. piperiformis-were merged into a single well-supported clade/MOTU. Although E. carmelinoi, E. evandroi and E. lenti were indistinguishable using coi DNA barcodes, E. piperiformis did form a distinct phylogenetic cluster and could be correctly identified using best-match criteria. Despite their apparent morphological differences, we propose on the basis of the molecular similarity of their DNA barcodes that these latter four morphospecies should be considered members of a recently-diverged species complex.

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Section: Introductionmentioning

confidence: 99%

Single‐locus DNA barcoding and species delimitation of the sandfly subgenus Evandromyia (Aldamyia)

Rodrigues¹,

Baton²,

Shimabukuro

2020

Medical Vet Entomology

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“…All these cases were compared to related species within the same genus, and all seemed to have unique barcode sequences – except for some cases of Nyssomyia and Trichophoromyia genera – which can be used for future molecular identification of these taxa. Some other studies have evaluated the usefulness of COI barcodes in the identification of sand flies from Central America [7, 41] and Colombia [9, 17, 42-43]. Moreover, in Colombia, the sampling efforts were carried out mainly in the Caribbean and Andean regions of the country, which can have different sand fly fauna compared with the southeast region [44].…”

Section: Discussionmentioning

confidence: 99%

Improving theCOIDNA barcoding library for Neotropical phlebotomine sand flies (Diptera, Psychodidae)

Posada-López

Rodrigues

Vélez

et al. 2022

Preprint

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A fragment of the mitochondrial cytochrome c oxidase subunit I (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 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 splitted into at least two 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 Ny. 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. Thus, the COI DNA barcode analysis allowed 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.

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“…This, in most of cases results in the impossibility of sand fly identification because of the damage of taxonomic value structures or inadequate material preparation, which does not affect molecular analysis at all. Secondly, most taxonomic keys allow the identification of sand fly species only during adult stage (Young and Duncan, 1994), whereas molecular studies can be done also from larval stages (Vivero et al, 2017). Thirdly, there are morphologically identical species that are reproductively isolated; hence, the traditional classification is unable to distinguish among them.…”

Section: Discussionmentioning

confidence: 99%

Sand fly typing: a simple and morphologically-supported method based on polymorphism of 18S rRNA gene in a Leishmaniasis endemic area of Argentina

et al. 2020

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Leishmaniases are vector-borne diseases that in the Americas are distributed from southern United States to northern Argentina. The vectors for this disease are small dipterans known as sand flies that are usually identified morphologically by observing structures with taxonomic value; but it is time-consuming, laborious, and requires entomological expertise. Then, this work was aimed at identifying sand flies with molecular techniques, using the morphological identification as a reference technique, in an endemic area of American Tegumentary Leishmaniasis (ATL) located in northern Argentina. For this, sand flies were caught at two patches of vegetation adjacent to rural areas in Orán department, Salta Province. Females were dissected with sterile needles; the head and last abdominal segments were analyzed for morphological identification. The remaining thorax and abdominal segments were used to extract DNA, which was amplified by PCR of the small subunit (SSU), 18S rRNA gene. PCR products were digested with CviQI and DdeI enzymes to identify sand fly species by Restriction Fragment Length Polymorphism (RFLP) analysis. Thus, the restriction pattern of each caught species was defined according to morphological identification. A total of 1501 females, belonging to four sand fly species, were captured. Nyssomyia neivai (1347/1501) was the most abundant species, followed by Migonemyia migonei (90/ 1501). From the total, 801 females were morphologically and molecularly identified, while 700 females were characterized only molecularly. For those females analyzed by both methods, there was total coincidence in the achieved result. Besides, the 5% (38/801) of females that could not be determined morphologically due to inadequate mounting were molecularly identified. All the females characterized just by PCR-RFLP, were successfully identified. Our results indicate that the explored method is capable of identifying the sand fly species that circulate in an ATL endemic area. Since this method is based on the analysis of markedly different patterns, the identification process might be more easily reproduced, as the bias introduced by the technician's lack of experience is removed.

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DNA barcode for identification of immature stages of sand flies (Diptera: Psychodidae) collected from natural breeding sites

Cited by 15 publications

References 30 publications

Single‐locus DNA barcoding and species delimitation of the sandfly subgenus Evandromyia (Aldamyia)

Single‐locus DNA barcoding and species delimitation of the sandfly subgenus Evandromyia (Aldamyia)

Improving theCOIDNA barcoding library for Neotropical phlebotomine sand flies (Diptera, Psychodidae)

Sand fly typing: a simple and morphologically-supported method based on polymorphism of 18S rRNA gene in a Leishmaniasis endemic area of Argentina

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