Abstract:Background
Lutzomyia
longipalpis
(Diptera: Psychodidae) is the major vector of
Leishmania
(Leishmania) infantum and thus plays a crucial role in the epidemiology of American visceral leishmaniasis (AVL). This vector is the best studied species of sand fly in the Neotropical region. Many studies claim that this vector is in fact a species complex; however there is still no consensus regarding the number of species that belong into this complex or the geographical distribution of sibling species. The aim o… Show more
“…A few studies have been carried out on microsatellites Watts et al, 2005;Zhang et al, 2009Zhang et al, , 2013Mahamdallie et al, 2010;Khalid et al, 2012;Santos et al, 2013) and the majority of studies focuses on the amplification of one or several molecular markers.…”
“…A few studies have been carried out on microsatellites Watts et al, 2005;Zhang et al, 2009Zhang et al, , 2013Mahamdallie et al, 2010;Khalid et al, 2012;Santos et al, 2013) and the majority of studies focuses on the amplification of one or several molecular markers.…”
“…2 and Table 1. Notable is the fact that chemotype 1 (9MGB) was also identified in Lutzomyia cruzi (Mangabeira, 1938) [50], a sibling species that may turn out to be a part of the L. longipalpis complex [51]. The possibility that they are isomers has not been investigated by more refined analyses (Hamilton JG, Brazil R., Personal communication, 2015).Fig.…”
Section: Introductionmentioning
confidence: 99%
“…Differential rates of evolution among genes, maintenance of ancestral polymorphisms and introgression are factors that describe discordant evolutionary history, especially for very closely related species such as L. longipalpis complex [59]. Microsatellites and nuclear genes associated with sexual behavior in Drosophila such as cacophony , paralytic and period ( per ) genes are among the markers that have proven useful for identifying clusters of putative species [47, 48, 51, 52, 57, 58, 60–63]. …”
In this paper we review the natural history of pheromone communication and the current diversity of aggregation-sex pheromones in the sand fly Lutzomyia longipalpis. This species complex is the main vector of Leishmania infantum, the agent of visceral leishmaniasis in the Americas. The identification of variation in pheromone chemotypes combined with molecular and sound analyses have all contributed to our understanding of the extent of divergence among cryptic members of this complex. The importance of chemical signals as pre-mating barriers and drivers of speciation is discussed. Moreover, the importance of aggregation-sex pheromones as sexually selected signals is highlighted with evidence from the literature suggesting their potential role in species and mate recognition as well as mate assessment. The distinct evolutionary forces possibly involved are briefly reviewed and discussed in the context of this intriguing insect.
“…The number of studies on the use of molecular markers, based on mitochondrial DNA (Folmer et al, 1994;Zhang and Hewitt, 1997), microsatellites (Queller et al, 1993;Santos et al, 2013) and, more recently, Single Nucleotide Polymorphism (SNPs) (Brumfield et al, 2003;Soria-Carrasco et al, 2014;Vignal et al, 2002), for the identification and evaluation of intra-and interpopulation diversity is increasing. Genotyping by traditional Sanger sequencing has also been used for the analysis of target regions but this approach proves costly for population genetics studies.…”
Standardization of the methods for extraction of DNA from sand flies is essential for obtaining high efficiency during subsequent molecular analyses, such as the new sequencing methods. Information obtained using these methods may contribute substantially to taxonomic, evolutionary, and eco-epidemiological studies. The aim of the present study was to standardize and compare two methods for the extraction of genomic DNA from sand flies for obtaining DNA in sufficient quantities for next-generation sequencing. Sand flies were collected from the municipalities of Campo Grande, Camapuã, Corumbá and Miranda, state of Mato Grosso do Sul, Brazil. Three protocols using a silica column-based commercial kit (ReliaPrep™ Blood gDNA Miniprep System kit, Promega), and three protocols based on the classical phenol-chloroform extraction method (Uliana et al., 1991), were compared with respect to the yield and quality of the extracted DNA. DNA was quantified using a Qubit 2.0 fluorometer. The presence of sand fly DNA was confirmed by PCR amplification of the IVS6 region (constitutive gene), followed by electrophoresis on a 1.5% agarose gel. A total of 144 male specimens were analyzed, 72 per method. Significant differences were observed between the two methods tested. Protocols 2 and 3 of phenol-chloroform extraction presented significantly better performance than all commercial kit extraction protocols tested. For phenol-chloroform extraction, protocol 3 presented significantly better performance than protocols 1 and 2. The IVS6 region was detected in 70 of 72 (97.22%) samples extracted with phenol, including all samples for protocols 2 and 3. This is the first study on the standardization of methods for the extraction of DNA from sand flies for application to next-generation sequencing, which is a promising tool for entomological and molecular studies of sand flies.
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