BackgroundDengue, chikungunya, malaria, filariasis and Japanese encephalitis are common mosquito-borne diseases endemic to Sri Lanka. Aedes aegypti and Aedes albopictus, the major vectors of dengue, were recently shown to undergo pre-imaginal development in brackish water bodies in the island. A limited survey of selected coastal localities of the Jaffna district in northern Sri Lanka was carried out to identify mosquito species undergoing pre-imaginal development in brackish and saline waters. The effect of salinity on the toxicity of Bacillus thuringiensis israelensis larvicide to Ae. aegypti larvae at salinity levels naturally tolerated by Ae. aegypti was examined.MethodsLarvae collected at the selected sites along the Jaffna coast were identified and salinity of habitat water determined in the laboratory. The LC50 and LC90 of B. thuringiensis toxin, the active ingredient of a commercial formulation of the larvicide BACTIVEC®, were determined with Ae. aegypti larvae. Bioassays were also carried out at salinities varying from 0 to18 ppt to determine the toxicity of Bacillus thuringiensis to fresh and brackish water-derived larvae of Ae. aegypti.ResultsLarvae of four Anopheles, two Aedes, one Culex and one Lutzia species were collected from brackish and saline sites with salinity in the range 2 to 68 ppt. The LC50 and LC90 of B. thuringiensis toxin for the second instar larvae of Ae. aegypti in fresh water were 0.006 ppm and 0.013 ppm respectively, with corresponding values for brackish water populations of 0.008 and 0.012 ppm respectively. One hundred percent survival of second instar fresh water and brackish water-derived Ae. aegypti larvae was recorded at salinity up to 10 and 12 ppt and 100% mortality at 16 and 18 ppt, yielding an LC 50 for salinity of 13.9 ppt and 15.4 ppt at 24 h post-treatment respectively for the two populations. Statistical analysis showed significantly reduced toxicity of B. thuringiensis to fresh and brackish water-derived Ae. aegypti larvae at high salinities.ConclusionA variety of mosquito vectors of human diseases undergo pre-imaginal development in brackish or saline waters in coastal areas of the Jaffna district in northern Sri Lanka. Salinity has a small but significant negative impact on the toxicity of B. thuringiensis toxin to Ae. aegypti larvae at salinity levels where Ae. aegypti larvae are found in the environment. This has implications for the use of B. thuringiensis toxin as a larvicide in brackish waters.

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BackgroundAnopheles annularis s.l. is a wide spread malaria vector in South and Southeast Asia, including Sri Lanka. The taxon An. annularis is a complex of two sibling species viz. A and B, that are differentiated by chromosome banding patterns and ribosomal gene sequences in India. Only species A is reported to be a malaria vector in India while the occurrence of sibling species in Sri Lanka has not been documented previously.FindingsAnopheline larvae were collected at a site in the Jaffna district, which lies within the dry zone of Sri Lanka, and reared in the laboratory. Emerged adults were identified using standard keys. DNA sequences of the D3 domain of 28S ribosomal DNA (rDNA) and the internal transcribed spacer-2 (ITS-2) of the morphologically identified An. annularis were determined. BLASTn searches against corresponding An. annularis sequences in GenBank and construction of phylogenetic trees from D3 and ITS-2 rDNA sequences showed that the Sri Lankan specimens, and An. annularis s.l. specimens from several Southeast Asian countries were closely related to species A of the Indian An. annularis complex.ConclusionsThe results show the presence of the malaria vector An. annularis species A in Sri Lanka and Southeast Asia. Because An. annularis vectors have been long associated with malaria transmission in irrigated agricultural areas in the Sri Lankan dry zone, continued monitoring of An. annularis populations, and their sibling species status, in these areas need to be integral to malaria control and eradication efforts in the island.

The correct identification of sand fly vectors of leishmaniasis is important for controlling the disease. Genetic, particularly DNA sequence data, has lately become an important adjunct to the use of morphological criteria for this purpose. A recent DNA sequencing study revealed the presence of two cryptic species in the Sergentomyia bailyi species complex in India. The present study was undertaken to ascertain the presence of cryptic species in the Se. bailyi complex in Sri Lanka using morphological characteristics and DNA sequences from cytochrome c oxidase subunits. Sand flies were collected from leishmaniasis endemic and non-endemic dry zone districts of Sri Lanka. A total of 175 Se. bailyi specimens were initially screened for morphological variations and the identified samples formed two groups, tentatively termed as Se. bailyi species A and B, based on the relative length of the sensilla chaeticum and antennal flagellomere. DNA sequences from the mitochondrial cytochrome c oxidase subunit I (COI) and subunit II (COII) genes of morphologically identified Se. bailyi species A and B were subsequently analyzed. The two species showed differences in the COI and COII gene sequences and were placed in two separate clades by phylogenetic analysis. An allele specific polymerase chain reaction assay based on sequence variation in the COI gene accurately differentiated species A and B. The study therefore describes the first morphological and genetic evidence for the presence of two cryptic species within the Se. bailyi complex in Sri Lanka and a DNA-based laboratory technique for differentiating them.

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