IPRC - 2018
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Item A Statistical Approach to Define Thresholds for Dengue Epidemic Management in Akurana Medical Officer of Health Area, Kandy District of Sri Lanka(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Udayanga, N.W.B.A.L.; Gunathilaka, P.A.D.H.N.; Iqbal, M.C.M.; Fernando, M.A.S.T.; Abeyewickreme, W.Stegomyia indices, namely; Premise Index (PI), Breteau Index (BI) and Container Index (CI) are used forvector management approaches in Sri Lanka. Properly defined threshold values for larval indices are of higher importance to provide forecasts on dengue epidemics and also for effective larval management of dengue vectors. However, such critical thresholds are poorly defined for Sri Lanka. The present study aimed to define threshold values forabove larval indices for dengue epidemic management in the Akurana Medical Officer of Health (MOH) in the Kandy District. Larval surveys were conducted on a monthly basis from January, 2016 to June, 2018. Four larval indices, namely BI for Aedesaegypti (BIA) and Aedesalbopictus (BIB), PI and CI were calculated. Further, monthly larval indices of AkuranaMOH area from January, 2012 to December, 2015, were obtained from the MOH office, along with monthly reported dengue cases for the entire study period. Receiver Operating Characteristic (ROC) curves in SPSS (version 23) were used to assess the discriminative power of the larval indices in determiningdengue epidemics and thresholds based on larval indices. As indicated by the area of ROC curve (AUC), the BIA (0.661) and PI (0.637) were having a notable discriminative power to forecast dengue epidemics at a two-month lag period. Both BIB (0.397) and CI (0.526) were non-informative influencers at one and two-month lag periods. The BIA and PI were better predictors of dengue incidence than BIB and CI. Based on the ROC curve, three risk thresholds were defined for BIA as Low Risk (BIA≤2.1), Moderate Risk (3.9≤BIA<4.85), and High Risk (BIA≥4.85), with respect to Ae. aegypti. According to the PI, thresholds were defined as Low Risk (PI≤6.2), Moderate Risk (7.7≤ PI<9.9), and High Risk (PI≥ 9.9). Threshold values defined for BI of Ae. aegypti and PI, could be recommended to be considered in implementing vector control efforts in the above study area for effective dengue epidemic management, through pre planned entomological management of dengue vectors.Item Laboratory Evaluation of the Bio-control Efficacy of Selected Copepods on Dengue Vectors of Sri Lanka(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Udayanga, N.W.B.A.L.; Ranathunge, R.M.T.; Iqbal, M.C.M.; Abeyewickreme, W.; Hapugoda, M.Among variety of alternatives for management of dengue vectors, biological control remains as a promising approach, due to its high efficacy, sustainability and low impacts on human and ecosystem health. Copepods are considered asa leading predator of mosquito larvae including Aedes vectors. However, the predatory potential of different copepods on bio-control of dengue vectors has been less studied in Sri Lanka. Therefore, the current study aimed to evaluate the predatory success of five locally abundant copepods on both AedesaegyptiandAe. albopictus. Copepod collections were made from water bodies and rock pools located in the Kandy and Gampaha districts by using a plankton net. After morphological identification, single gravid copepods of different species were used to establish copepod cultures under standard laboratory conditions. Five adult copepods of each species were transferred into containers and groups of 200 first instar Ae. Aegypti larvae were introduced in to each container, separately. The number of surviving larvae in each container was recorded at 3 hour intervals up to 24 hours. Five replicates were conducted for each copepod species. Same experimental design was followed for Ae. albopictus. General Linear Modelling technique (GLM) followed by Tukey’s pair-wise comparison was used to make statistical inferences on the significance of average larvalconsumption rates by studied copepod species. SPSS (version 23) was used for the statistical analysis. Five species of copepods, namely Cyclops languides, C. varicans, C. vernalis, Mesocyclopleuckarti and M. scrassus were considered during the study. Average predation rates of 34.9±1.80 and 33.5±1.06 for Ae. aegypti and Ae. albopictus, respectively, were indicated by M. leuckartias the highest predation rates. Meanwhile, M. scrassus showed the second highest predation rates for both Aedes vectors. On the other hand, the lowest predatory efficacy was shown by C. languides with 10.6+1.60 and 8.4+1.10 for Ae. aegypti and Ae. albopictus, respectively. The average consumption rates of different copepod species varied significantly (p<0.05 at 5% level of significance). The type of Aedes species had a significant influence on the predatory efficacy of studied copepods (p< 0.05) under laboratory conditions. In conclusion, M. leuckartiand M. scrassus that reported the highest predation efficacies on both Aedes larvae could be suggested as potential bio-control agents for dengue vector management approaches in Sri Lanka after semi field and field settings.Item Sterile Insect Technique for Controlling Aedes albopictus (Skuse); Sex Separation and Mating Competitiveness(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Harishchandra, J.; Hapugoda, M.; Premaratne, R.G.; Abeyewickreme, W.Sterile Insect Technique (SIT) introduced by Edward F. Knipling in 1950s has been shown as an effective and sustainable genetic approach to control populations of selected pest insects in area wide integrated pest management programmes. Successfulness of this approach depends on the availability of an efficient sexing method and production of sterile males competitive enough to compete with wild males. In the context of developing SIT for the control of dengue vector Aedes albopictus (Skuse) in Sri Lanka, a method for separation of male pupae was tested and mating competitiveness of irradiated males against wild males was evaluated under laboratory conditions. Male pupae of the F42 generation of Ae. albopictus (Gampaha strain) were separated from female pupae using mechanical sieving method. A set of three stainless steel sieves of mesh sizes 1.4, 1.25 and 1.12mm were placed one on the other in descending order of size towards the bottom, forming a column immersed in a water jar. All pupae were poured on to the top sieve and shaken gently for seven minutes. Pupae were collected from each sieve and the male and female counts were taken after emergence of adults. Male and female pupae used for the competitiveness assessment were separated by the above sieving method. Male pupae (F42, age 28-40hours) were exposed to Gamma radiation (Gamma 220, Atomic Energy of Canada Ltd., Co60) at a dose of 50Gy by placing 300 male pupae in each plastic vial with 10ml of water. Dosimetry was performed using Frickey method. Wild fertile males and females reared from field-collected eggs from Gampaha (F1) were used to compete and cross with irradiated males respectively in different ratios. Two days after the emergence, the adult mosquitoes were caged (Bug Dorm 30X30X30cm) with ratios of irradiated males: wild males: wild virgin females 1:1:1, 3:1:1 and 5:1:1 (females, n = 100). Fertile control was set with wild males and wild virgin females 1:1. and an irradiated control was set with irradiated males with wild virgin females 1:1. Mosquitoes were allowed to mate for 3 days with access to 10% sugar solution. Then the mosquitoes were fed with cattle blood and engorged females were isolated in oviposition tubes. After 4 days, females were returned into cages and the filter papers containing eggs were allowed to dry. Eggs were hatched after one week and the hatching rates were compared. From the pupae retained in the sieve of mesh size 1.12mm, 98.3% were males. The mean hatch rates of fertile and irradiated controls and with ratios of irradiated males: wild fertile males: wild virgin females 1:1:1, 3:1:1, 5:1:1 were 80.24%±1.63SEM, 2.42%±0.57SEM, 56.74%±5.79SEM, 37.33%±4.76SEM and 34.23%±4.73SEM respectively. Fried Competitive Index (C) of irradiated males compared to controls was 0.43. Results show sieving method can be used for separating male pupae for irradiation with 98% accuracy. Irradiated males were less than 50% competitive as wild males under laboratory conditionsItem Potential Use of a Combined Sterile Insect Technique (SIT) and Wolbachia-Based Approach for the Control of the Dengue Vector Aedes albopictus in Sri Lanka(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Wijegunawardana, N.D.A.D.; Chandrasena, T.G.A.N.; Abeyewickreme, W.; Dassanayake, R.S.; Xi, Z.; Bourtzis, K.; Gunawardene, Y. I. N.S.This study addresses the possibility of using a novel and environmental friendly strategy of combined sterile insect technique (SIT) and a Wolbachia-based approach in Aedes albopictus population control efforts in Sri Lanka. The above strategy combines irradiation and the intracellular endosymbiont bacteria Wolbachia, in insects which are vertically transmitted from mother to offspring. Releasing sterile male mosquitoes in the wild will introduce sterility for suppression of Ae. albopictus wild mosquito population. The strategy aims at the releasing of sexually incompatible Wolbachia triple infected and irradiated male mosquitoes on a large scale to the natural environment. By integrating irradiation, any accidentally released female mosquito will be made fully sterile, and hence the risk of population replacement of the target population with triple infected mosquitoes is eliminated. Therefore, the combined SIT/IIT approach offers a safe and bio-secure strategy for the population suppression of Ae. albopictus in Sri Lanka. The overall work involved assessment of the density and species abundance of wild mosquito populations, introgression, and characterization of Wolbachia triple infected Ae. albopictus line in a Sri Lankan genetic background and determination of minimum sterility dose for the males and females of the introgressed line. Results revealed that Ae. albopictus mosquito was the dominant vector species in the study area (Narangodapaluwa PHI area) with an average ovitrap index of 75.25% (range; 56.9% - 94.7%) over the study year. The frequency of Wolbachia infection among the mosquito population was ~17% (13/78). The most prominent Wolbachia super group infecting the wild mosquito species was the B group and Ae. albopictus was naturally infected with both strains A and B. Eighty-five new alleles and 8 complete allelic profiles were submitted to the Wolbachia MLST database. All sequencing data were deposited in the Genebank under the accession numbers MH447376-MH470292 and MH756095-MH777430. Successful transfer of Wolbachia triple infection to Ae. albopictus into Sri Lankan genetic background was achieved through an introgression experiment and characterization of the newly developed Ae. albopictus line expressed partial CI and revealed competitive fitness cost for fecundity, fertility, and longevity compared with wild-type while there was no cost for pupation rate, adult emergence and sex ratio. Therefore, integration of an irradiation (dose; 30Gy) is recommended to achieve complete male and female sterility prior to mosquito release (minimum sterility dose for males - 28 Gy and females - 30 Gy). This minimum sterility dose is lower than the irradiation dose applied for SIT and had not resulted in a significant effect on male competitiveness. Therefore, a combined SIT and Wolbachia based approach in Ae. albopictus dengue vector control is recommended as an additional tool in integrated vector management in Sri Lanka.