Biology, bionomics and life-table studies of Anopheles stephensi (Diptera: Culicidae) in Sri Lanka and estimating the vectorial potential using mathematical approximations.

BACKGROUND: Anopheles stephensi is an invasive mosquito in Sri Lanka that can potentially transmit malaria. The transmission intensity is linked with biology, bionomic and behavioral aspects of a vector that are associated with the Vectorial Capacity (VC). However, the influence of larval conditions eventually affects the vectorial potential of An. stephensi are not well understood. METHODS: A colony of An. stephensi was established at the Regional Centre of the Open University of Sri Lanka, Jaffna District. The colony was maintained under confined conditions according to standard protocols. Biotypes of An. stephensi were characterized by referring to the number of egg ridges. Information on (a) biological aspects of eggs (duration for egg hatching, egg development and hatchability), (b) larval development time, larval survivorship pupation success, resting depth of larvae), (c) pupae (adult emergence rate, average time for adult emergence) and (d) adults (biting frequency, mating success gonotrophic cycle, fecundity, duration for egg-laying, percentage of sexes, adult survival/longevity) were evaluated under life-table analysis. Further, selected morphometric characters of each life cycle stage were recorded from the eggs (length and breadth), larvae (head length, width of head, length of thorax, width of thorax, length of abdomen, width of abdomen, and the total length of larvae), pupae (cephalothoracic length and width) and adults (length & width of wing, thorax and abdomen). The VC was calculated using a mathematical-based approach. Descriptive statistics, General Linear Model (GLM) and independent-sample t-test were used for the statistical analysis. RESULTS: All three biotypes were identified based on egg morphology. Mysorensis biotype (47%; n = 470) was predominant followed by type (38.1%; n = 381) and intermediate (14.9%; n = 149). The mean egg length (F(2,997) = 3.56; P = 0.029) and breadth (F(2,997) = 4.57; P = 0.011) denoted significant differences among the three biotypes. The mating success of females observed was 80.7 ± 4.45%. The mean hatching period was 1.9 ± 0.03 days, with a hatching rate of 86.2 ± 0.77%. Overall, 8.0 ± 0.14 days were required for larval development and 30.3 ± 0.14 h were spent in the pupal stage. The pupation success was 94.5 ± 0.37%, and the majority were males (53.1 ± 0.73%). The mean fecundity was 106.5 ± 6.38 eggs and a gonotrophic cycle of 3.4 ± 0.06 days. The female survival rate was 43.2 ± 2.4%, with a mean biting frequency of 66.6 ± 3.5%. The average VC of adult An. stephensi was estimated to be 18.7. CONCLUSIONS: The type biotype, which is an effective vector in the Indian subcontinent is present in Sri Lanka. According to the mathematical approximation, An. stephensi found locally has a vectorial capacity of over 18. Therefore, this study warrants the health authorities and vector control programmes to continue the entomological surveys, monitoring of vector densities and implementing appropriate vector control interventions based on biology and bionomic information of vectors.

Comparative analysis of the larvicidal activity of temephos (EC50) and novaluaron (EC10) to control Anopheles stephensi in Sri Lanka.

BACKGROUND: Anopheles stephensi was first recorded in the coastal area of Mannar District, Sri Lanka, in December 2016. Since then, this vector has been isolated from other districts in the Northern and Eastern Provinces of Sri Lanka. Chemical control is the main arm of vector control that can be used to reduce the vector densities within a short period. Thus, the present study aimed at evaluating the efficacy of using selected insecticides for the control of An. stephensi larvae. METHOD: The third and fourth instar larval stages of An. stephensi (F2 generation) of field mosquitoes that were caught using cattle baited net trap collections from Columbuthurai, Kurunagar, and Navanthurai areas in Jaffna District, Sri Lanka, were obtained from the laboratory colony established at Jaffna. Batches of 100 larvae were taken for experiments and introduced separately to a concentration series of temephos and novaluron (0.04-400 ppm). A control test was also performed at each setup without introducing insecticides. The mortality rates of An. stephensi larvae exposed to different concentrations of larvicides were recorded at 1, 24 and 48-h intervals. The experiment was replicated five times at individual concentrations for each selected chemical. Data were analyzed using the General Linear Model (GLM) and Probit analysis. RESULTS: The highest mortality rate (100%) at a 1-h exposure period was observed from temephos at >100 ppm. The mortality rates varied significantly for different concentrations and larvicides (p < 0.05). At 24-h of the exposure period, the 100% mortality of An. stephensi larvae were observed from both temephos and novaluron even at 0.04 ppm. CONCLUSION: Both temephos and novaluron reported 100% mortality rates in An. stephensi larvae at 1-h and 24-h exposure periods. Based on the findings, temephos and novaluron can be recommended as effective larvicides for chemical-based control of An. stephensi in Jaffna, Sri Lanka. Further, it is recommended to conduct a field-based study, where habitat types and water quality are highly heterogeneous and may affect the residual activity.

Developmental responses and survival of Anopheles stephensi larval stages at different salinity levels.

BACKGROUND: Anopheles stephensi is a newly invaded vector in Sri Lanka. It has been identified in coastal areas in the northern and eastern parts of the country and evidences the ability to breed in brackish water environments. METHODS: Laboratory investigations were conducted with batches of 100 first and third instar larvae exposed to a salinity gradient (0-40 ppt). Survival rates at 1 h, 24 h and until pupation were recorded for first and third instar larvae at different salinity levels. The experiment was repeated four times for both instars. Data were analysed using the general linear model and probit analysis. RESULTS: Significant variations in adult emergence were observed from both larval stages at different salinity levels (p<0.05). The highest pupation rates were observed at 2.5 ppt salinity. The survival rate of first instar larvae after 24 h of salinity exposure was >80% up to 12.5 ppt, while 100% mortality was observed for from the ≥25 ppt level of salinity. More than 90% of the third instar larvae pupated from salinity levels <15 ppt. The lowest survival rate was reported as 15.8±2.47% at 25 ppt. CONCLUSIONS: This indicates a high potential of increasing density of A. stephensi in coastal ecosystems in lagoons and other saline water bodies. Hence it is high time to redesign vector control interventions for vector breeding in coastal ecosystems.