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In recent decades, the consumption of tea has increased due to its various associated health benefits, prompting growing concerns regarding the safety and quality of tea products. Nevertheless, there has been a significant dearth of scientific information regarding the microbiological status of black tea leaves. This study sought to bridge this knowledge gap by investigating the microflora present in tea leaves at various processing stages of black tea production. Samples were meticulously collected during distinct processing steps, and their culturable microorganisms were identified through sequence‐based methods. The results revealed that the predominant bacterial genus throughout the tea manufacturing process was Bacillus, constituting a substantial 77% of the identified bacterial population. Other leading genera included Klebsiella (8%), Lysinibacillus (4%), Escherichia (2%) with the remaining 9% comprising various other genera. Among the fungal community, Aspergillus and Penicillium species exhibited a significantly higher relative abundance, each comprising 24%. Furthermore, yeast communities included Debaryomyces, Candida, Hyphopichia, Rhodosporidiobolus, and Wickerhamomyces species. Lactobacillus fermentum was identified in the fermented tea leaves, highlighting its role in the fermentation process. Yeasts and molds were also present in the final tea product, indicating potential post‐processing contamination. The study did not detect any mycotoxins in any of the samples. These findings had indicated that presence of some microorganisms is extremely common in different processing stages while alien microorganisms are being introduced during manufacturing. Thus, emphasizing the need for stringent regulations and quality assurance practices within the tea industry to ensure the safety and quality. [ABSTRACT FROM AUTHOR]

Background: Aedes aegypti and Ae. albopictus are important vectors of human diseases such as dengue, chikungunya, and zika. In Sri Lanka, they have been responsible for transmitting dengue virus. One of the most important parameters influencing the likelihood of arbovirus transmission is the age structure of the mosquito population. However, mosquito age is difficult to measure with accuracy. This study aims to construct multivariate calibration models using the transcriptional abundance of three age-responsive genes: Ae15848 (calcium-binding protein), Ae8505 (structural component of cuticle), and Ae4274 (fizzy cell cycle/cell division cycle 20). Methods: The transcriptional age-grading technique was applied to determine the chronological age of Ae. aegypti and Ae. albopictus female mosquito populations from Sri Lanka using the age-responsive genes Ae15848, Ae8505, and Ae4274. Furthermore, Ae. aegypti samples obtained from colonies reared at two temperatures (23 and 27 °C) were used to investigate the influence of temperature on this age-grading technique. Expression levels of these three genes were quantified using reverse transcription qualitative PCR (qRT-PCR), and results were normalized against the housekeeping gene ribosomal gene S17 (RpS17). Results: The expression of Ae15848 and Ae8505 decreased with the age of mosquitoes and showed the most significant and consistent change while expression of Ae4274 increased with age. The multivariate calibration models showed > 80% correlation between expression of these age-responsive genes and the age of female mosquitoes at both temperatures. At 27 °C the accuracy of age predictions using the models was 2.19 (± 1.66) days and 2.58 (± 2.06) days for Ae. aegypti and Ae. albopictus females, respectively. The accuracy of the model for Ae. aegypti at 23 °C was 3.42 (± 2.74) days. Conclusions: An adult rearing temperature difference of 4 °C (23–27 °C) did not significantly affect the age predictions. The calibration models created during this study could be successfully used to estimate the age of wild Ae. aegypti and Ae. albopictus mosquitoes from Sri Lanka. [ABSTRACT FROM AUTHOR]

Background: Leishmania donovani-induced and sand fly-transmitted leishmaniasis is a growing health problem in Sri Lanka. Limited knowledge on biological and behavioral characteristics of probable vector Phlebotomus argentipes hinders disease control. Here, insecticide susceptibility patterns of P. argentipes were investigated with exploration of probable underlying resistance mechanisms. Methods: Adult sand flies were collected using standard cattle baited net traps and CDC light traps from selected sites in four districts. Adult F1 progeny of P. argentipes were exposed to different concentrations of DDT, malathion, deltamethrin and propoxur using WHO susceptibility bioassay kits. Post-1-h knockdown and post-24-h mortality were recorded and analyzed. Metabolic enzyme activity and the sensitivity of the acetylcholinesterase target-site were determined by biochemical assays using wild-caught flies. Extracted fly DNA samples were tested for the presence of knockdown-resistance (kdr) type mutations. Results: The LC100 values for DDT, malathion, propoxur and deltamethrin were 0.8–1.5%, 0.9–2.0%, 0.017–0.03% and 0.007% respectively. Insecticide-susceptibility levels were higher than the discriminating dosages established for Aedes mosquitoes, except for malathion. The lowest susceptibility levels (except for deltamethrin) were detected in the Mamadala population, whereas the highest levels were detected in the Mirigama population. The percentage of knocked-down sand flies was < 75% at any tested concentration, including those, which exhibited 100% mortality after 24 h. Elevated activity levels of glutathione S-transferase (3%, 7%, 12.5% and 14%) and esterase (2%, 5%, 5.5% and 6.5%) were detected in flies that originated from Mirigama, Pannala, Thalawa and Mamadala respectively, while monooxygenase quantities remained below the cut-off level. Ten to 34.5% of flies were heterozygous for acetylcholinesterases target-site insensitivity, associated with organophosphate and carbamate resistance. Pyrethroid-resistance-associated L1014F kdr-type mutation in the voltage gated sodium channel gene was detected in 30/53 flies. Conclusions: Populations of P. argentipes in Sri Lanka are largely susceptible to common insecticides, except for malathion (used extensively in the past for malaria control). Their insecticide susceptibility appears negatively associated with past malaria endemicity of the study sites, with signs of early insecticide tolerance. Presence of insecticide target site insensitivity in a notable proportion of flies and enhanced insecticide metabolizing enzyme activities imply potential future challenges for leishmaniasis control, with a call for urgent proactive measures for its containment. [ABSTRACT FROM AUTHOR]

Cryptosporidium is a protozoan parasite that causes gastroenteritis in both humans and animals. Cryptosporidium infections in humans and animals have been reported from many parts of Sri Lanka. During the present study, five international electronic databases were extensively searched for peer-reviewed research papers published on the prevalence of Cryptosporidium infections and their occurrence in probable sources in Sri Lanka, within the time frame from 1987 to 2023. Collected information revealed that Cryptosporidium oocysts are more commonly found in surface water compared to well water, and shallow wells have a higher occurrence than deep wells. Contamination of river water with Cryptosporidium is mainly from the faecal matter coming from various domestic, agricultural and wildlife animal species. In Sri Lanka, Cryptosporidium infections in captive non-human primates is possibly due to contaminated food and human interaction. In wild animals, infections are more common in species that feed on the ground, suggesting contaminated soil and water as the source of infection. Most of the infected animals are asymptomatic, and many had co-infections with other enteric parasites. Molecular analysis of Cryptosporidium samples from infected primates revealed the presence of four major clades of C. parvum, with some isolates closely related to zoonotic C. parvum genotypes. Information on the habitats of the infected primates suggests that livestock is the primary source of infection. Several studies have been conducted to investigate the prevalence of Cryptosporidium infection in various livestock animals, particularly in goats, cattle, buffaloes, and swine in Sri Lanka. These studies reveal high prevalence of asymptomatic Cryptosporidium infection in goats and the potential for the transmission of zoonotic C. parvum from goats, cattle and buffaloes to humans. Molecular epidemiological analysis identified new genotypes of Cryptosporidium in domestic bovids (cattle and water buffalo), with no evidence of the commonly reported zoonotic species C. parvum. Waterborne transmission is the most common mode of infection of Cryptosporidium that affect both humans and animals. Studies conducted in human populations are primarily based in hospitals and pre-schools, and conclude that Cryptosporidium infections are a common cause of diarrhoea in children under the age of five. In conclusion, Cryptosporidium infections occur in different species of animals and humans in Sri Lanka and the oocysts have been detected in surface water, which might be an important source of infection for animals and humans in the country. [ABSTRACT FROM AUTHOR]

Vector resistance to insecticides is a significant challenge in the control of mosquito-borne diseases such as dengue. Analysis of the growth rate and survivorship of the immature stages of larvae has been important in formulating effective vector control strategies. The present study aimed to assess insecticide resistance in both larvae and adults of Aedes aegypti and Ae. albopictus, and the life tables of their immature stages. Mosquito eggs collected from Kurunegala and Colombo districts of Sri Lanka from January to March 2023 were reared under laboratory conditions. Larval bioassays were carried out with temephos (0.0125 ppm, 0.0625ppm and 0.125ppm) and, the adult bioassays were with pyrethroids (0.03% deltamethrin, 0.05% and 0.08% lambdacyhalothrin, 0.04% permethrin) and malathion (1%and 5%). Biochemical assays were used to evaluate the activity of insecticide-detoxifying enzymes and the insensitivity of the organophosphate target site acetylcholinesterase. Survivorship of immature stages under laboratory conditions and semi-field conditions i.e., bamboo stumps, tyres and clay pots, were studied using Kaplan-Meier survival analysis. Larvae and adults of both species from Kurunegala were susceptible to all the insecticides tested, whereas Colombo populations were resistant. Acetylcholinesterases were largely sensitive in all vector populations. Higher activities of esterases, glutathione s-transferases and monooxygenases were found in Colombo populations. Results indicated that Ae. aegypti larvae were more resistant to insecticides than Ae. albopictus larvae. Also, the mechanisms of insecticide resistance were well developed in adults than in larvae. Aedes aegypti larval development time (5 days) was significantly shorter than that of Ae. albopictus (9 days) under laboratory conditions. Survival probabilities of them were 76-80%. Egg hatching to adult emergence period was 8 days for Ae. aegypti and 13 days for Ae. albopictus. This period, under semifield conditions were; bamboo stumps 7-7.2 days, tyres 10-11.3 days, and clay pots 13.8-14 days with survival rates varied from 28.8 to 63% for both species. This study highlights stage specific response of dengue vectors to insecticides and survivorship of their immature stages in different habitats. This knowledge can be effectively used in future dengue vector control programmes. [ABSTRACT FROM AUTHOR]

Summary Culex tritaeniorhynchus Giles and Cx. gelidus Theobald (Diptera: Culicidae), both vectors of Japanese encephalitis, were collected in 1984 and 1998 from two disease endemic localities in Sri Lanka: Anaradhapura and Kandy. Using wild-caught adult mosquitoes from light traps, log dosage-probit mortality curves for insecticide bioassays were obtained for three insecticides: malathion (organophosphate), propoxur (carbamate) and permethrin (pyrethroid). LD50 values showed that, in 1998, Cx. tritaeniorhynchus was ∼100-fold more resistant to malathion and 10-fold more resistant to propoxur than was Cx. gelidus. This difference was attributed to Cx. tritaeniorhynchus breeding mostly in irrigated rice paddy fields, where it would have been exposed to pesticide selection pressure, whereas Cx. gelidus breeds in other types of aquatic habitats less prone to pesticide applications. Resistance in Cx. tritaeniorhynchus increased between 1984 and 1998, whereas Cx. gelidus remained predominantly susceptible. Propoxur inhibition of acetylcholinesterase (AChE) activity (the target site of organophosphates and carbamates) indicated that in 1998, frequencies of insensitive AChE-based resistance were 9% in Cx. gelidus and 2–23% in Cx. tritaeniorhynchus, whereas in 1984 this resistance mechanism was detected only in 2% of the latter species from Anaradhapura. The AChE inhibition coefficient (ki) with propoxur was 1.86 ± 0.24 × 105 m-1 min-1 for Cx. tritaeniorhynchus from Anaradhapura in 1998. Both species were tested for activity levels of detoxifying glutathione S-trans- ferases (GSTs) and malathion-specific as well as general carboxylesterases. High activities of GSTs and carboxylesterases were detected in Cx. tritaeniorhynchus but not Cx. gelidus. Malathion-specific carboxylesterase was absent from both species. Native polyacrylamide gel electrophoresis resolved two elevated general carboxylesterases, CtrEstβ1 and CtrEstα1, from Cx. tritaeniorhynchus and none from Cx. gelidus. CtrEstβ1 was the most intensely staining band. Gel inhibition experiments showed that both elevated esterases were inhibited by organophosphates and carbamates but not by pyrethroids. The major elevated esterase CtrEstβ1 was partially purified (15-fold) by sequential Q-Sepharose and phenyl Sepharose column chromatography. The bimolecular rate constant (ka) and the deacylation rate constant (k3) for the malaoxon/ enzyme interaction were 9.9 ± 1.1 × 103 m-1 min-1 and 3.5 ± 0.05 × 10-4 m-1 min-1, respectively, demonstrating that the role of this enzyme in organophosphorus insecticide resistance is sequestration. [ABSTRACT FROM AUTHOR]

Insecticide-resistant strains of the brown planthopper, Nilaparvata lugens , from four locations all had a single diffuse elevated esterase band on native polyacrylamide gel electrophoresis. The white backed planthopper Sogatella furcifera had two elevated esterases with lower relative mobilities than the N. lugens esterase, while the insecticide-susceptible N. bakeri and the grass-associated sibling species of N. lugens sensu lato all had a single low intensity staining esterase with a lower relative mobility than the resistance associated N. lugens esterase. All the esterases were inhibited by pre-incubation with 0.1 mM paraoxon, but were not affected by permethrin up to its solubility limit, indicating their possible role in organophosphorus, but not pyrethroid insecticide, resistance. Partial purification of the elevated esterase from insecticide resistant Sri Lankan N. lugens showed that despite its diffuse nature on gels it purified as a single isoform, with a specific activity of 5.85 mumol min- 1 mg- 1 and an estimated molecular weight of approximately 60 kDa. Insecticide resistance was conferred by this elevated esterase through rapid binding and slow turnover of the carbamate or the insecticidally active oxon analogues of the phosphorothioates, i.e. sequestration rather than metabolism is the primary resistance mechanism. In contrast to earlier studies on the elevated esterases of N. lugens from Japan and the Philippines, we were unable to detect any malathion metabolism by this esterase, nor did malathion inhibit the esterase, although it was sensitive to inhibition by malaoxon. The elevated N. lugens esterase did not cross-react with antisera raised to the elevated mosquito or aphid esterases. The efficacy of the partially purified N. lugens esterase in insecticide sequestration and turnover is compared with other well characterized amplified esterases from mosquitoes and aphids. [ABSTRACT FROM AUTHOR]

Two sibling species of Nilaparvata lugens sensu lato were collected from cultivated rice (Oryza sativa) and two from the wild grass Leersia hexandra . Strains of the rice-associated species of N. lugens were from populations exposed to insecticide selection pressure over an extended period, while the Leersia -associated species had little prior insecticide exposure. The rice-associated species from Batalagoda, Sri Lanka and Sulawesi, Indonesia both showed low (2- 5-fold) levels of resistance to malathion and propoxur, as measured by increased times to knock-down, and 2- 17-fold resistance to permethrin at the LT50 level compared with the Leersia -associated species from Bogor, Indonesia. The Goan Leersia -associated species had similar levels of susceptibility to malathion, but significantly lower susceptibility to permethrin than the Bogor one. Resistance in the Sri Lankan rice strain of N. lugens increased significantly after malathion selection in the laboratory. The major insecticide resistance mechanism in the two rice-associated strains was based on elevation of esterase activity. A highly elevated esterase electromorph was visible on native polyacrylamide gels of crude homogenates of these strains compared with those from Leersia . The diffuse nature of the esterase band suggested that it may contain more than one electromorph, but a range of electrophoretic techniques failed to differentiate more than one variant. Esterase activity increased after insecticide selection of the Sri Lankan rice-associated strain, while no effect on other possible resistance-associated enzyme systems was observed. The increased level of permethrin tolerance in the Goan Leersia strain was synergized by the monooxygenase inhibitor piperonyl butoxide. This synergist had no effect on the toxicity of permethrin or propoxur to the Sri Lankan rice-associated strain of N. lugens . There was no evidence of altered acetylcholinesterase or glutathione S -transferase based resistance mechanisms in any of the strains tested. [ABSTRACT FROM AUTHOR]

SummaryElevated esterase Estβ1 was purified from larvae of newly isolated strains of the mosquito Culex quinquefasciatus from Colombia (COL) and Trinidad (TRI) with resistance to organophosphate (OP) insecticides. Insecticide interactions were compared with those of elevated Estβ12 from the OP-resistant Habana strain and the non-elevated Estβ13 from the susceptible PelSS strain. On the basis of insecticide binding efficiency, all elevated Estβ1 esterases were readily distinguishable. Differences between the EcoRI restriction fragment patterns of the amplified estβ1 gene in COL and TRI strains compared with each other, and between amplified estβ11, estβ12 and the non-amplified estβ13, suggest differences in their nucleotide sequence. Considering their variable insecticide binding efficiencies, these genetic differences would imply that, in contrast to estα2 and estβ2, amplification of estβ1 has occurred several times independently. Generally, the elevated Estβ1s were more reactive with insecticides than the non-elevated Estβ13. This supports the hypothesis that the elevated esterase-based mechanism confers resistance through amplification of alleles coding for esterases which have a greater specificity for the insecticides they sequester than the esterases coded by their non-amplified counterparts. [ABSTRACT FROM AUTHOR]

Elimination of vector mosquito larvae and their breeding environments is an effective strategy in dengue disease control. Present study examined larval density and water quality in breeding habitats and container preference of dengue vectors Ae. aegypti and Ae. albopictus. Larval surveys were conducted monthly in urban, semiurban, and rural sites in Kurunegala, Sri Lanka, from January 2019 to December 2021. Larval densities were recorded under the following three categories: type of container (16 types), type of material (6 types), and location (indoor/outdoor). Breeding preference ratios (BPRs) were calculated using Index of Available Containers and the Index of Contribution to Breeding Sites. Out of 19,234 wet containers examined, larval stages were found in 1,043 habitats. Ae. albopictus larvae were in all three areas whereas Ae. aegypti larvae were restricted to urban areas. Highest number of wet containers and highest positivity were reported from urban followed by semiurban. In general, discarded nondegradable items were the most frequent and mostly positive breeding sites. For Ae. aegypti, the most preferred breeding sites were gutters and concrete slabs. Ae. albopictus mostly preferred concrete slabs in urban areas and tyres in semiurban and rural areas. Material types such as rubber and concrete were mostly preferred by Ae. aegypti whereas ceramic was preferred by Ae. albopictus. Although plastic was the most available material type in all study sites, preference to plastic was low except for urban Ae. albopictus. Both species preferred urban indoor breeding habitats although outdoor breeding was preferred by Ae. albopictus in rural areas. Larval densities of Ae. aegypti and semiurban Ae. albopictus significantly correlated with the BPR of the container type and material type. Dengue vector larvae were found in a 6.7–9.4 pH range. Total dissolved solids and alkalinity positively correlated with preference. Information generated can be successfully used in waste management and public education for effective vector control. [ABSTRACT FROM AUTHOR]

Insect Growth Regulator (IGR) novaluron is an alternative to synthetic neuro-inhibitory insecticides. Present study was designed to assess appropriate dosages of novaluron for dengue vector control. Larvae of Aedes aegypti and Ae. albopictus were exposed to a concentration series of novaluron (Rimon EC10) for two fixed exposure periods of 7-days and 14-days to determined LC50 and LC99 values. Inhibition of adult emergence (IE50 and IE99) was determined by a 14-day exposure. Semi-field experiments were conducted by exposing cohorts of Ae. aegypti larvae to IE99, 2 × IE99 and 10 × IE99 novaluron concentrations in water storage buckets (10 L) and plastic barrels (200 L). For the 7-day exposure, LC50 values were 0.047–0.049 ppm and LC99 were 0.144–0.151 ppm. For 14-day exposure, these values were 0.002–0.005 ppm and 0.006–0.01 ppm respectively. For both species, IE99 was 0.001 ppb under semi-field conditions, and was effective for nearly 2 months. Novaluron concentration 0.01 ppb was effective up to 3 months, with an IE of 89–95%. Authorities should critically review a reduction of the presently recommended field dosage of 200 ppm novaluron by × 100 or more. This would provide the same efficacy but mitigate environmental pollution, development of vector resistance, and financial losses. [ABSTRACT FROM AUTHOR]

We investigated the physicochemical properties and the biotic interactions of breeding sites of tropical mosquito species. Field sampling was done in 12 study areas in Sri Lanka covering areas with secondary natural forests and human settlements. A total of 226 breeding sites were investigated to determine the biotic interactions and physiochemical properties of breeding water (pH, Conductivity, Dissolved Oxygen, Total Dissolved Solids and Temperature). A total of 80.5% of breeding sites from both habitats were positive for mosquito larvae of seven genera and 24 species. Orthopodomyia flavithorax (297) and Aedes albopictus (295) were dominated in tree holes of Alstonia macrophylla, Vateria copallifera and Artocarpus nobilis. Diversity indices showed that the diversity of mosquitoes is high in wet zone habitats of Sri Lanka compared to dry and intermediate zone habitats. Aedes albopictus coexisted with 11 different mosquito species while it avoided larvae of Culex fuscanus, Cx. uniformis and Tripteroides affinis. Strong positive associations were reported between Ae. albopictus and Ar. subalbatus while larvae of Or. flavithorax mosquitoes were not co‐occurred with the larvae of Ae. vittatus, Ae. aegypti, Cx. sitiens, Ar. subalbatus, Anopheles spp and Tr. affinis. The findings identified the breeding adaptability and tolerance to a wide range of physiochemical properties of tropical mosquito communities. [ABSTRACT FROM AUTHOR]

Background: Wolbachia are a group of maternally inherited intracellular bacteria known to be widespread among arthropods. Infections with Wolbachia cause declines of host populations, and also induce host resistance to a wide range of pathogens. Over the past few decades, researchers were curious to use Wolbachia as a biological tool to control mosquito vectors. During the present study, assessment of the prevalence of Wolbachia infections among wild mosquito populations in Sri Lanka where mosquito-borne diseases are a major health concern, was carried out for the first time. DNA was extracted from the abdomens of mosquitoes, collected from seven provinces, and screened for the presence of Wolbachia by PCR using wsp and groE primers. Group-specific and strain-specific primers were used to classify Wolbachia into the supergroups A and B, and into the strains Mel, AlbA and Pip. Results: A total of 330 individual mosquitoes belonging to 22 species and 7 genera were screened. Eighty-seven mosquitoes (26.36%) belonging to four species (i.e. Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Mansonia uniformis) were positive for Wolbachia infections. Primary vector of the dengue fever, Ae. aegypti was negative for Wolbachia infections while the secondary vector, Ae. albopictus, showed a very high infection rate. The filarial vector C. quinquefasciatus had a relatively high rate of infection. Japanese encephalitis vectors C. gelidus and C. triteaneorynchus, and the Anopheles vectors of malaria were negative for Wolbachia infections. Nine sequences of Wolbachia-positive PCR products were deposited in the GenBank and compared with other available data. Aedes albopictus was infected with both Wolbachia strains A (AlbA) and B (Pip) supergroups. Phylogenetic analysis of the wsp sequences showed two major branches confirming identities obtained from the PCR screening with strainspecific primers. Conclusion: Wolbachia infections were found only among four mosquito species in Sri Lanka: Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Mansonia uniformis. Sequence data showed high haplotype diversity among the Wolbachia strains. [ABSTRACT FROM AUTHOR]

Dengue is an important vector-borne disease transmitted by the mosquitoes Aedes aegypti and Ae. albopictus. In the absence of an effective vaccine, vector control has become the key intervention tool in controlling the disease. Vector densities are significantly affected by the changing weather patterns of a region. The present study was conducted in three selected localities, i.e., urban Bandaranayakapura, semiurban Galgamuwa, and rural Buluwala in the Kurunegala district of Sri Lanka to assess spatial and temporal distribution of dengue vector mosquitoes and to predict vector prevalence with respect to changing weather parameters. Monthly ovitrap surveys and larval surveys were conducted from January to December 2019 and continued further in the urban area up to December 2021. Aedes aegypti was found moderately in the urban area and to a lesser extent in semiurban but not in the rural area. Aedes albopictus had the preference for rural over urban areas. Aedes aegypti preferred indoor breeding, while Ae. albopictus preferred both indoor and outdoor. For Ae. albopictus, ovitrap index (OVI), premise index (PI), container index (CI), and Breteau index (BI) correlated with both the rainfall (RF) and relative humidity (RH) of the urban site. Correlations were stronger between OVI and RH and also between BI and RF. Linear regression analysis was fitted, and a prediction model was developed using BI and RF with no lag period (R2 (sq) = 86.3%; F = 53.12; R2 (pred) = 63.12%; model: Log10 (BI) = 0.153 + 0.286 ∗ Log10 (RF); RMSE = 1.49). Another prediction model was developed using OVI and RH with one month lag period (R2 (sq) = 70.21%; F = 57.23; model: OVI predicted = 15.1 + 0.528 ∗ Lag 1 month RH; RMSE = 2.01). These two models can be used to monitor the population dynamics of Ae. albopictus in urban settings to predict possible dengue outbreaks. [ABSTRACT FROM AUTHOR]

Background: The current status of insecticide resistance and the underlying resistance mechanisms were studied in the major vector of malaria, Anopheles culicifacies, and the secondary vector, Anopheles subpictus in five districts (Anuradhapura, Kurunegala, Moneragala, Puttalam and Trincomalee) of Sri Lanka. Eight other anophelines, Anopheles annularis, Anopheles barbirostris, Anopheles jamesii, Anopheles nigerrimus, Anopheles peditaeniatus, Anopheles tessellatus, Anopheles vagus and Anopheles varuna from Anuradhapura district were also tested. Methods: Adult females were exposed to the WHO discriminating dosages of DDT, malathion, fenitrothion, propoxur, λ-cyhalothrin, cyfluthrin, cypermethrin, deltamethrin, permethrin and etofenprox. The presence of metabolic resistance by esterase, glutathione S-transferase (GST) and monooxygenase-based mechanisms, and the sensitivity of the acetylcholinesterase target site were assessed using synergists, and biochemical, and metabolic techniques. Results: All the anopheline species had high DDT resistance. All An. culicifacies and An. subpictus populations were resistant to malathion, except An. culicifacies from Kurunegala, where there was no malathion carboxylesterase activity. Kurunegala and Puttalam populations of An. culicifacies were susceptible to fenitrothion. All the An. culicifacies populations were susceptible to carbamates. Both species were susceptible to the discriminating dosages of cypermethrin and cyfluthrin, but had different levels of resistance to other pyrethroids. Of the 8 other anophelines, only An. nigerrimus and An. peditaeniatus were resistant to all the insecticides tested, probably due to their high exposure to the insecticides used in agriculture. An. vagus showed some resistance to permethrin. Esterases, GSTs and monooxygenases were elevated in both An. culicifacies and An. subpictus. AChE was most sensitive to insecticides in Kurunegala and Trincomalee An. culicifacies populations and highly insensitive in the Trincomalee An. subpictus population. Conclusion: The complexity of the resistance segregating in these field populations underlines the need for new molecular tools to identify the genomic diversity, differential upregulation and different binding specificities of resistance conferring genes, and the presence of different subspecies with different vectorial capacities. [ABSTRACT FROM AUTHOR]

Phlebotomus argentipes is the vector of Leishmania donovani which causes the disease leishmaniasis, a neglected tropical disease and a growing health problem in Sri Lanka. A proper understanding of the population genetic structure of sand fly vectors is considered important prior to planning and implementation of a successful vector control program. Thus, the present study was conducted to determine the population genetic structure of sand fly vectors in Sri Lanka. Two mitochondrial genes namely Cytochrome c oxidase subunit 1 (Cox 1) and Cytochrome b (Cytb), and the internal transcribed spacer 2 (ITS2) region from the nuclear ribosomal DNA were used for molecular characterization. Analyses included maximum likelihood method, network analysis and DNA polymorphisms. The outcome revealed unique sequences of all genomic regions studied except the cox 1 gene had a relationship with sand flies isolated previously from Sri Lanka, India and Israel and cytb gene of 4 sand flies that aligned with those isolated earlier from Sri Lanka and 3 from Madagascar. Furthermore, cox 1 gene and ITS 2 region analyses based on FST values indicated a possible gene flow between the study sites whereas cytb gene analysis favoured the existence of genetically distinct populations of P. argentipes in each of the study sites. Poor population differentiation of P. argentipes, a possible consequence of a gene flow, is indeed of concern due to the risk imposed by promoting the spread of functionally important phenotypes such as insecticide resistance across the country, making future vector control efforts challenging. [ABSTRACT FROM AUTHOR]

Pyrethroid resistance has imposed a severe threat to the control programmes of dengue vectors Aedes aegypti and Ae. albopictus in Sri Lanka. Present study investigated the resistance status and underlying mechanisms of these mosquitoes in six provinces of Sri Lanka. Mosquitoes were collected from Badulla (Uva Province), Batticaloa (Eastern), Colombo (Western), Jaffna (Northern), Kandy (Central) and Kurunegala (Northwestern). Adults were exposed to discriminating dosages of permethrin, deltamethrin, λ-cyhalothrin, cyfluthrin and DDT according to WHO guidelines. Survivals from bioassays were tested for kdr type mutations. Elevation of insecticide metabolizing enzymes and insensitivity of acetylcholinesterases, the target site of organophosphates and carbamates, were also tested. A total of 4875 mosquitoes (2687 Ae. aegypti, 2188 Ae. albopictus) were subjected to bioassays. All the populations had high resistance to DDT though it has not been used in the country for more than four decades. In general, Ae. aegypti populations showed more resistance towards pyrethroids. Glutathione S-transferase (GST) activities were enhanced in most of the populations with no enhancement of carboxylesterase and monooxygenase activities. Acetylcholinesterases were not altered in both species. Aedes aegypti showed high resistance to pyrethroids, mainly due to three kdr type mutations; V1016I, S989P and F1534C found in the voltage gated sodium channel which is the target site of pyrethroids and DDT. Both species were highly resistant to DDT mainly through a GST based mechanism. In the absence of elevated esterase-based and insensitive AChE mechanisms, organophosphates and carbamates can be recommended as alternatives for future dengue vector control programmes in Sri Lanka. [ABSTRACT FROM AUTHOR]

Avian blood parasites have been shown to have significant health effects on avifauna worldwide. Sri Lanka, a tropical island rich with resident and migratory birds, has not been properly evaluated for avian blood parasites or their vectors. We investigated the presence of avian haemoparasites in Sri Lankan birds and the potential mosquito vectors of those pathogens. Blood samples were collected from local/migratory birds captured by standard mist nets from Anawilundawa bird sanctuary, Hanthana mountain range, and the University of Peradeniya park. Mosquitoes were collected from Halgolla forest reserve and the forest patches in Kurunegala and Gampola areas in addition to the above mist-netting localities. Part of the mitochondrial cytochrome b (cytb) gene was amplified and sequenced to detect the presence of haemoparasites from avian blood samples (86) and mosquito samples (480). Blood parasites of the two genera, i.e., Haemoproteus (4 species; Haemoproteus sp. 1-4) and Plasmodium (5 species; Plasmodium sp. 1-5) were identified from seven bird species (four resident and three migratory). Among these, three bird species (Red-vented bulbul (3/16), Asian Brown flycatcher (1/1), and India pitta (1/1)) were positive for Plasmodium spp., while four (Yellow-browed bulbul (1/4), oriental white-eye (1/4), brown-headed Barbet (1/4), and Indian blue robin (1/1)) were positive for Haemoproteus spp. Two mosquito species were also positive for Plasmodium (3) and Haemoproteus (1) species. Phylogenetic analysis and haplotype networks created using positive sequences of haemoparasites showed that a Plasmodium clade was shared by Cx nigropunctatus mosquitoes and the migratory bird, Indian pitta. The majority (85%) of the Plasmodium and Haemoproteus sequences of this study were not linked to the well-characterized species suggesting the distinct nature of the lineages. Associations between mosquito species and blood parasites of birds suggest the possible vector status of these mosquitoes. [ABSTRACT FROM AUTHOR]

1. Resource partitioning is a critical component in competing species that coexist in a community. The biting behaviour of coexisting frog‐biting mosquito species associated with a tropical anuran community is investigated. 2. Monthly samplings were taken for 2 years at two study sites in central Sri Lanka to collect frog‐biting mosquitoes, anuran abundance, environmental data, and interactions between mosquitoes and anuran hosts. Mosquitoes were collected using mouth‐operated aspirators and sound traps broadcasting anuran calls. Mosquitoes were identified using taxonomic keys and DNA barcodes. 3. A total of 1079 frog‐biting mosquitoes from four species belonging to two genera were collected [Uranotaenia rutherfordi (5%), Ur. morphotype 1 (67%), Ur. morphotype 2 (21%), and Mansonia uniformis (7%)]. Species‐specific interactions between Uranotaenia mosquitoes and their anuran host were found. Uranotaenia morphotype 1, the most common species, was mainly attracted (99%) to Duttaphrynus melanostictus. Uranotaenia rutherfordi was mainly attracted (95%) to Pseudophilautus rus, while Ur. morphotype 2 was attracted (97%) to Polypedates cruciger. These Uranotaenia species are active at different hours at night that correspond to the peak calling activity of their anuran host. Each Uranotaenia species was active at heights that coincide with the calling sites of their host. In contrast, Ma. uniformis was non‐specific in host choice and was equally distributed in space and time with respect to host feeding. 4. Here, previously unknown feeding patterns of co‐occurring frog‐biting mosquito species and their interactions with anurans present in their community are reported, highlighting the existence of complex behavioural patterns of these mosquito communities. [ABSTRACT FROM AUTHOR]

Tropical agricultural soils have been claimed as a source of carbon. As agricultural systems in the tropics are highly diverse, it is useful to study soil organic C (SOC) of different agricultural systems. We quantified the SOC fractions, available nutrients, and aggregate stability in eight different tropical agricultural systems, including annual crops under different management scenarios, such as organic, inorganic, and combined fertilizer applications. Annual crops treated with organic fertilizer only (A–OF), inorganic fertilizer only (A–IF), both organic and inorganic fertilizers (A–O/IF), perennial crops (PC), home gardens (HG), and abandoned home gardens (AHG) in Eutrustox soils and annual crops with organic fertilizer only (A–OFS) and uncultivated land on Quartzipsamments soil (USR) were studied. The links between SOC fractions, available nutrients, and aggregate stability in these soils were analyzed. Regression models were fitted for SOC fractions and available nutrients. Our results indicated that the different land use types exhibited significant variations in organic carbon fractions, aggregate stability, and available nutrients in soils. The available macro and micronutrients, except for nitrogen, showed a significant positive correlation with either total organic C (TOC) or carbon fractions indicating the synergy between them. The differences in soil C stocks clearly reflected the differences in litter fall and soil disturbance, as indicated by the highest C stocks in AHG. The dry weight of collected litter showed that AHG accumulated the highest litter content (97.38 g/m2) compared to the lowest (37.63 g/m2) in A–I/F. Organic matter addition to soil also increased the C stocks, even in annual crops. Aggregate stability showed a positive correlation with C fractions. The regression models developed in this study can be used to predict available nutrients by measuring TOC or C fractions in similar land use types in the tropics. This study confirmed that tropical agricultural systems that include annual crops have potential for storing and maintaining C in soils, if appropriately managed. The beneficial influence of SOC on available nutrients and aggregate stability could be a driving force to increase carbon stock in tropical agricultural systems. [ABSTRACT FROM AUTHOR]

Background: Vectors of mosquito-borne diseases in Sri Lanka, except for malaria, belong to the subfamily Culicinae, which includes nearly 84% of the mosquito fauna of the country. Hence, accurate and precise species identification of culicine mosquitoes is a crucial factor in implementing effective vector control strategies. During the present study, a combined effort using morphology and DNA barcoding was made to characterize mosquitoes of the subfamily Culicinae for the first time from nine districts of Sri Lanka. Cytochrome c oxidase subunit 1 (cox1) gene from the mitochondrial genome and the internal transcribed spacer 2 (ITS2) region from the nuclear ribosomal DNA were used for molecular characterization. Results: According to morphological identification, the field collected adult mosquitoes belonged to 5 genera and 14 species, i.e. Aedes aegypti, Ae. albopictus, Ae. pallidostriatus, Aedes sp. 1, Armigeres sp. 1, Culex bitaeniorhynchus, Cx. fuscocephala, Cx. gelidus, Cx. pseudovishnui, Cx. quinquefasciatus, Cx. tritaeniorhynchus, Cx. whitmorei, Mansonia uniformis and Mimomyia chamberlaini. Molecular analyses of 62 cox1 and 36 ITS2 sequences were exclusively comparable with the morphological identifications of all the species except for Ae. pallidostriatus and Aedes sp. 1. Although the species identification of Armigeres sp. 1 specimens using morphological features was not possible during this study, DNA barcodes of the specimens matched 100% with the publicly available Ar. subalbatus sequences, giving their species status. Analysis of all the cox1 sequences (14 clades supported by strong bootstrap value in the Neighbor-Joining tree and interspecific distances of > 3%) showed the presence of 14 different species. This is the first available DNA sequence in the GenBank records for morphologically identified Ae. pallidostriatus. Aedes sp. 1 could not be identified morphologically or by publicly available sequences. Aedes aegypti, Ae. albopictus and all Culex species reported during the current study are vectors of human diseases. All these vector species showed comparatively high diversity. Conclusions: The current study reflects the significance of integrated systematic approach and use of cox1 and ITS genetic markers in mosquito taxonomy. Results of DNA barcoding were comparable with morphological identifications and, more importantly, DNA barcoding could accurately identify the species in the instances where the traditional morphological identification failed due to indistinguishable characters of damaged specimens and the presence of subspecies. [ABSTRACT FROM AUTHOR]

Background: Anopheles stephensi, the major vector of urban malaria in India, was recently detected for the first time in Sri Lanka in Mannar Island on the northwestern coast. Since there are different biotypes of An. stephensi with different vector capacities in India, a study was undertaken to further characterise the genotype and biotype of An. stephensi in Mannar Island. Methods: Mosquito larvae were collected in Pesalai village in Mannar and maintained in the insectary until adulthood. Adult An. stephensi were identified morphologically using published keys. Identified adult An. stephensi were molecularly characterized using two mitochondrial (cox1 and cytb) and one nuclear (ITS2) markers. Their PCR-amplified target fragments were sequenced and checked against available sequences in GenBank for phylogenetic analysis. The average spiracular and thoracic lengths and the spiracular index were determined to identify biotypes based on corresponding indices for Indian An. stephensi. Results: All DNA sequences for the Mannar samples matched reported sequences for An. stephensi from the Middle East and India. However, a single nucleotide variation in the cox1 sequence suggested an amino acid change from valine to methionine in the cox1 protein in Sri Lankan An. stephensi. Morphological data was consistent with the presence of the Indian urban vector An. stephensi type-form in Sri Lanka. Conclusions: The present study provides a more detailed molecular characterization of An. stephensi and suggests the presence of the type-form of the vector for the first time in Sri Lanka. The single mutation in the cox1 genemay be indicative of a founder effect causing the initial diversification of An. stephensi in Sri Lanka from the Indian form. The distribution of the potent urban vector An. stephensi type-form needs to be established by studies throughout the island as its spread adds to the challenge of maintaining the country's malaria-free status. [ABSTRACT FROM AUTHOR]

Background: Genus Anopheles is a major mosquito group of interest in Sri Lanka as it includes vectors of malaria and its members exist as species complexes. Taxonomy of the group is mainly based on morphological features, which are not conclusive and can be easily erased while handling the specimens. A combined effort, using morphology and DNA barcoding (using the markers cytochrome c oxidase subunit I (COI) gene and internal transcribed spacer 2 (ITS2) region, was made during the present study to recognize anophelines collected from eight districts of Sri Lanka for the first time. Methods: Cytochrome c oxidase subunit I and ITS2 regions of morphologically identified anopheline mosquitoes from Sri Lanka were sequenced. These sequences together with GenBank sequences were used in phylogenetic tree construction and molecular characterization of mosquitoes. Results: According to morphological identification, the field-collected adult mosquitoes belonged to 15 species, i.e., Anopheles aconitus, Anopheles annularis, Anopheles barbirostris, Anopheles culicifacies, Anopheles jamesii, Anopheles karwari, Anopheles maculatus, Anopheles nigerrimus, Anopheles pallidus, Anopheles peditaeniatus, Anopheles pseudojamesi, Anopheles subpictus, Anopheles tessellatus, Anopheles vagus, and Anopheles varuna. However, analysis of 123 COI sequences (445 bp) (16 clades supported by strong bootstrap value in the neighbour joining tree and inter-specific distances of >3%) showed that there are 16 distinct species. Identity of the morphologically identified species, except An. subpictus, was comparable with the DNA barcoding results. COI sequence analysis showed that morphologically identified An. subpictus is composed of two genetic entities: An. subpictus species A and species B (inter-specific K2P distance 0.128). All the four haplotypes of An. culicifacies discovered during the present study belonged to a single species. ITS2 sequences (542 bp) were obtained for all the species except for An. barbirostris, An. subpictus species B, An. tessellatus, and An. varuna. Each of these sequences was represented by a single species-specific haplotype. Conclusions: The present study reflects the importance and feasibility of COI and ITS2 genetic markers in identifying anophelines and their sibling species, and the significance of integrated systematic approach in mosquito taxonomy. Wide distribution of malaria vectors in the country perhaps indicates the potential for re-emergence of malaria in the country. [ABSTRACT FROM AUTHOR]

Background: The identification of species B and E in the Anopheles culicifacies complex in the Indian subcontinent has been based on Y-chromosome karyotype. Since no detectable variations were previously found in DNA markers commonly used for sibling species identification, further molecular characterization using cytochrome oxidase subunit I (COI) and microsatellite markers was carried out on Y-chromosome karyotyped Anopheles culicifacies specie B and E from Unnichchai, Kallady and Ranawarunawa in Sri Lanka. Findings: COI sequence analysis (n = 22) revealed the presence of nine unique haplotypes with six in each species. Three haplotypes were shared by both species. The two sibling species had a pairwise FST value of 1.338 (p < 0.05) with the number of migrants (Nm) value <1. The genetic structure analysis resulted in two genetic clusters not 100 % associated with karyotypes. While none of the species B were incorrectly assigned two were inconclusive. Five out of 26 specimens karyotyped as species E were incorrectly assigned, while further 9 were inconclusive. Conclusions: The new molecular data support the existence of two genetically different populations of the Culicifacies Complex in Sri Lanka that are not associated with the Y-chromosome karyotype. Detailed analysis with more microsatellite markers and assortative mating experiments are needed to establish the presence of the two genetically distinct populations and relate them to Y-chromosome morphology. [ABSTRACT FROM AUTHOR]

Unprecedented incidences ofidengue have been reported in Sri Lanka in recent years. The district of Batticaloa, which was devastated by the 2004 Asian tsunami, is one of the districts affected by dengue. One option to curtail this disease is to implement appropriate vector control measures. A nine-month study was carried out within the Batticaloa Municipal Council limit from April to December 2008. Larval collections were conducted fortnightly using conventional ovitraps for nine months covering the dry and wet seasons. Ovitraps (indoor and outdoor) were placed in 15 randomly selected houses. The collected larvae were brought to the laboratory and reared under laboratory conditions. The larval forms and emerged adults were identified on the basis of reported morphological descriptions. The identified adults of 2-3 d old were exposed to common insecticides following the WHO protocol. During the study period, a total of 10,685 Aedes aegypti and Ae. albopictus mosquitoes were collected, with the former constituting 57% of the total sample. Both species were collected from indoor and outdoor ovitraps, and their prevalence was recorded throughout the study period. A seasonal shift was observed in the density, with Ae. aegypti predominating during the dry season and Ae. albopictus during the wet season. Both species were highly resistant to 4% DDT and susceptible to 0.25% permethrin. The continuous presence of potential dengue vectors may have contributed to the dengue prevalence in the district. Since both species can oviposit in indoor and outdoor ovitraps, public awareness and participation should be promoted in the vector control programme of the Ministry of Health along with continuous vector surveillance. [ABSTRACT FROM AUTHOR]

DDT and pyrethroid resistance in Culex quinquefasciatus have been previously reported in Sri Lanka, but the mechanisms involved have yet to be characterized. We report the presence of two mutant alleles of the sodium channel gene, the target site for both DDT and pyrethroid insecticides. Both mutations resulted in classic knockdown resistance (kdr) L1014F mutation because of either an A-to-T substitution or an A-to-C substitution. We developed two alternative assays to distinguish between the two mutations and used these to screen 214 individuals from nine geographic locations throughout Sri Lanka. Very high levels of kdr mutations were found throughout the country. A predominance of the A-to-C mutation was observed over the A-to-T with an average allele frequency of 50% and 2%, respectively. In addition to these non-synonymous kdr substitutions, we also found an indel (TCACA) in the intron downstream of the kdr mutation. After genotyping this indel in 136 individuals, we found no evident correlation between kdr genotypes and intronic indel. The presence of two alternative kdr mutations has implications for the reliance on single molecular diagnostics for detection of resistance in field populations. Furthermore, the high levels of these kdr mutations in C. quinquefasciatus populations throughout Sri Lanka are of concern for the future of pyrethroid-based control programmes on this island. [ABSTRACT FROM AUTHOR]

The mosquito Culex quinquefasciatus poses a substantial threat to human and veterinary health as a primary vector of West Nile virus (WNV), the filarial worm Wuchereria bancrofti, and an avian malaria parasite. Comparative phylogenomics revealed an expanded canonical C. quinquefasciatus immune gene repertoire compared with those of Aedes aegypti and Anopheles gambiae. Transcriptomic analysis of C. quinquefasciatus genes responsive to WNV, W. bancrofti, and non-native bacteria facilitated an unprecedented meta-analysis of 25 vector-pathogen interactions involving arboviruses, filarial worms, bacteria, and malaria parasites, revealing common and distinct responses to these pathogen types in three mosquito genera. Our findings provide support for the hypothesis that mosquito-borne pathogens have evolved to evade innate immune responses in three vector mosquito species of major medical importance. [ABSTRACT FROM AUTHOR]

Background: Aedes aegypti were found developing in the water in open public drains (drain-water, DW) in Jaffna city in northern Sri Lanka, a location where the arboviral diseases dengue and chikungunya are endemic. Methods: Susceptibilities to the common insecticides dichlorodiphenyltrichloroethane (DDT), malathion, propoxur, permethrin and deltamethrin and activities of the insecticide-detoxifying enzymes carboxylesterase (EST), glutathione S-transferase (GST) and monooxygenase (MO) were compared in adult Ae. aegypti developing in DW and fresh water (FW). Results: DW Ae. aegypti were resistant to the pyrethroids deltamethrin and permethrin, while FW Ae. aegypti were susceptible to deltamethrin but possibly resistant to permethrin. Both DW and FW Ae. aegypti were resistant to DDT, malathion and propoxur. Greater pyrethroid resistance in DW Ae. aegypti was consistent with higher GST and MO activities. Conclusions: The results demonstrate the potential for insecticide resistance developing in Ae. aegypti adapted to DW. Urbanization in arboviral disease-endemic countries is characterized by a proliferation of open water drains and therefore the findings identify a potential new challenge to global health. [ABSTRACT FROM AUTHOR]

Background: The tropical bed bug, Cimex hemipterus, is a serious indoor public health pest in tropical regions causing intense physical discomfort and mental distress to humans. At present, the application of insecticides is the major control strategy. The present study was designed to evaluate the development of resistance and resistance mechanisms in Cimex hemipterus from Kandy district, Sri Lanka. Methods: The resistance status of the collected bed bugs was determined against the discriminative dosages of DDT, malathion, propoxur, deltamethrin and permethrin by conducting bioassays according to World Health Organization guidelines. Activities of insecticide metabolizing enzymes, i.e. esterases, glutathione S-transferases (GST) and monooxygenases, and the insensitivity of organophosphate/carbamate target site acetylcholinesterase (AChE), were evaluated by biochemical assays. Regions of the gene of the pyrethroid/DDT target site, the voltage-gated sodium channel regulatory protein (VGSC), were sequenced for possible kdr mutations. Results: Survival percentages of bed bug population were 71, 68 and 51% for DDT, malathion and propoxur respectively. KT50 and KT90 values, calculated using log-probit mortality curves for deltamethrin were 62.55 and 123.96 h, respectively. These values were much higher for permethrin where KT50 was 201.10 h and the KT90 was beyond the detectable range. Results were compared with previous values reported for the same population in 2002. Resistance to propoxur has increased significantly from 11 to 51% with about a 20-fold increase in the number of individuals with elevated esterase mechanism. No significant change has occurred in malathion and DDT resistance, in GST and monooxygenase activities, and in AChE sensitivity for the past 14 years. Six kdr associated mutations (Y/L995H, V1010L, I1011F, L1014F, V1016E, L1017F/S) and a non-kdr associated mutation (A1007S mutation) were found from the α-region of the VGSC gene. Out of the kdr type mutations, only L1014F has been reported previously form C. hemipterus while the others have been reported from other insects. Conclusions: The bed bug population has developed high resistance to propoxur with increased esterase activities. KT50 for deltamethrin and permethrin has increased 125- and 20-fold, respectively, over the period 2002 to 2016. To the authors' knowledge, this is the first time that the possible involvement of a kdr type mutation in developing pyrethroid resistance in C. hemipterus has been shown in Sri Lanka. [ABSTRACT FROM AUTHOR]

Background: Sri Lanka has been malaria-free since 2013 but re-introduction of malaria transmission by infected overseas travelers is possible due to a prevalence of potent malaria vectors. Knowledge of the insecticide resistance status among Anopheles vectors is important if vector control has to be reintroduced in the island. The present study investigated the insecticide susceptibility levels and resistance mechanisms of Anopheles sundaicus (sensu lato) (previously classified as Anopheles subpictus species B) an important malaria vector in the Jaffna Peninsula and it surroundings in northern Sri Lanka after indoor residual spraying of insecticides was terminated in 2013. Results: Species-specific PCR assays identified An. sundaicus (s.l.) in four locations in the Jaffna and adjacent Kilinochchi districts. Bioassays confirmed that An. sundaicus (s.l.) collected in Kilinochchi were completely susceptible to 0.05% deltamethrin and 5% malathion and resistant to 4% dichlorodiphenyltrichloroethane (DDT), whereas those from Jaffna were relatively susceptible to all three insecticides. Kilinochchi populations of An. sundaicus (s.l.) showed significantly higher glutathione S-transferase activity than population from Jaffna. However, Jaffna An. sundaicus (s.l.) had significantly higher Propoxur-resistant acetylcholinesterase activity. Activities of non-specific esterases and monooxygenases were not significantly elevated in An. sundaicus (s.l.) collected in both districts. Conclusions: The susceptibility to malathion and deltamethrin in An. sundaicus (s.l.) suggests that they can be still used for controlling this potential malaria vector in the Jaffna Peninsula and adjacent areas. Continuing country-wide studies on other malaria vectors and their insecticide susceptibilities are important in this regard. [ABSTRACT FROM AUTHOR]

Background: Although Sri Lanka is considered as a malaria-free nation, the threat of re-emergence of outbreaks still remains due to the high prevalence and abundance of malaria vectors. Analysis of population genetic structure of malaria vectors is considered to be one of the vital components in implementing successful vector control programmes. The present study was conducted to determine the population genetic structure of three abundant malaria vectors; Anopheles subpictus sensu lato (s.l.) , Anopheles peditaneatus and Anopheles vagus from five administrative districts in two climatic zones; intermediate zone (Badulla and Kurunegala districts) and dry zone (Ampara, Batticoloa and Jaffna districts) of Sri Lanka using the mitochondrial gene, cytochrome c oxidase subunit I ( COI ). Methods: Adult mosquitoes of An. subpictus s.l. , An. peditaeniatus, and An. vagus were collected from five study sites located in five districts using cattle baited traps and backpack aspirators. Representative samples of each species that were morphologically confirmed were selected from each locality in generating COI sequences (> 6 good quality sequences per species per locality). Results: Anopheles subpictus s.l. specimens collected during the study belonged to two sibling species; An. subpictus 'A' (from all study sites except from Jaffna) and An. subpictus 'B' (only from Jaffna). The results of haplotype and nucleotide diversity indices showed that all the three species are having high genetic diversity. Although a high significant pairwise difference was observed between An. subpictus ' A' and 'B' ( F st > 0.950, p < 0.05), there were no significant genetic population structures within An. peditaeniatus , An. vagus and An. subpictus species A ( p > 0.05), indicating possible gene flow between these populations. Conclusions: Gene flow among the populations of An. peditaeniatus , An. vagus and An. subpictus species A was evident. Application of vector control measures against all mosquito species must be done with close monitoring since gene flow can assist the spread of insecticide resistance genes over a vast geographical area. [ABSTRACT FROM AUTHOR]