Genetic characterization of Aedes aegypti (Diptera: Culicidae) in Sri Lanka based on COI gene.

BACKGROUND & OBJECTIVES: Aedes aegypti is the most prominent vector for dengue virus worldwide. Accurate identification of the species and understanding its colonization pattern are essential prerequisites in vector control. Thus, the present study was aimed to genetically characterize Ae. aegypti mosquitoes collected from different regions of Sri Lanka based on mitochondrial COI gene. METHODS: Thirty-three Ae. aegypti larval samples were collected from 19 districts. A 735bp region of the mitochondrial COI gene was amplified and analyzed for genetic diversity indices. Phylogenetic trees were constructed using Sri Lankan samples and also including mosquito samples reported from other parts of the world. RESULTS: High genetic diversity was observed within the samples analysed (gene diversity: 0.949; average number of nucleotide differences: 6.371). There were 20 haplotypes presented within the 19 localities investigated. The phylogenetic tree derived two main clades. However, no distinguishable clustering pattern was observed in the phylogenetic tree except for the districts in the northern corner indicating extensive admixing among different populations. When samples from other countries were included in the phylogenetic tree, Anuradhapura, and Mannar samples were clustered together with samples from India, Venezuela, USA, Portugal and Cambodia while Rathnapura was clustered with Bolivia and France. INTERPRETATION & CONCLUSION: Our results suggest that Sri Lanka has undergone multiple invasions of Ae. aegypti from various parts of the world over an extensive period. Further, the mosquito control campaigns had not caused a significant effect on the Ae. aegypti populations which is existing in mutation-drift equilibrium.

Anti-inflammatory and analgesic activities of mature fresh leaves of Vitex negundo.

This study confirmed the oral anti-inflammatory, analgesic and antihistamine properties of mature fresh leaves (MFL) of Vitex negundo L. (Verbenaceae) claimed in the Ayurveda medicine by orally treating a water extract of the leaves to rats. The early phase (2h) of carrageenan-induced rat paw oedema was significantly (P<0.01) suppressed in an inversely does-dependent (r(2)=1, P<0.01) manner by MFL. The EC(50) was 2g/kg of MFL. In the formaldehyde-induced rat paw oedema test, the 2.5 and 5g/kg leaves significantly (P<0.05) suppressed the inflammation on days 4-6 of the test. In the hot plate test, 2.5 and 5g/kg of MFL showed a significant (P<0.05) and directly dose-dependent analgesic activity at 1h of treatment while the activity was absent in the tail flick test in rats. The EC(50) for the analgesic activity was 4.1g/kg. In the formalin test, 1.25, 2.5 and 5g/kg of MFL significantly (P<0.05) suppressed the pain in both the phases of the test like aspirin. The leaves showed an inversely dose-dependent in vivo antihistamine and in vitro prostaglandin (PG) synthesis inhibition, membrane stabilising and antioxidant activities. Naloxone did not abolish the analgesic activity in the hot plate test. A 5g/kg of MFL did not impair muscle strength and co-ordination and did not induce sedation. The treatment of 5g/kg of MFL did not show signs of acute toxicity or stress. Fourteen-day oral treatment of 5g/kg of MFL significantly increased the serum activity of AST. Flowering of the tree did not abolish the analgesic and anti-inflammatory activities of the leaves. These observations revealed that the fresh leaves of Vitex negundo have anti-inflammatory and pain suppressing activities possibly mediated via PG synthesis inhibition, antihistamine, membrane stabilising and antioxidant activities. The antihistamine activity can produce the anti-itching effect claimed in Ayurveda medicine.