Graduate Studies
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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 Role of Larval Indices on the Incidence of Dengue Epidemics in Akurana, Medical Officer of Health (MOH) Area, Kandy.(In: Proceedings of the International Postgraduate Research Conference 2017 (IPRC – 2017), Faculty of Graduate Studies, University of Kelaniya, Sri Lanka., 2017) Udayanga, N.W.B.A.L.; Gunathilaka, P.A.D.H.N.; Iqbal, M.C.M.; Fernando, M.A.S.T.; Abeyewickreme, W.As in many other Asian countries, vector control entities in Sri Lanka also depend upon larval indices as the major driving factor of routine entomological surveillance and management of dengue vectors. However, studies aimed at defining the role of larval indices, which reflect the population dynamics of dengue vectors, on the incidence of dengue outbreaks are limited. This is mainly due to restrictions in long-term vector density related data. Hence, the current study attempts to evaluate the role of larval indices on the incidence of dengue epidemics in Akurana, Medical Officer of Health (MOH) area.Item Optimization of Critical Threshold Values for Aedes Mosquitoes based on Breteau Index in Kandy District of Sri Lanka.(In: Proceedings of the International Postgraduate Research Conference 2017 (IPRC – 2017), Faculty of Graduate Studies, University of Kelaniya, Sri Lanka., 2017) Udayanga, N.W.B.A.L.; Gunathilaka, P.A.D.H.N.; Iqbal, M.C.M.; Kusumawathie, P.H.D.; Abeyewickreme, W.Many countries, including Sri Lanka, mainly depend upon Stegomyia indices,such as Premise Index (PI), Breteau Index (BI),Container Index (CI), and Pupal Index (PI) for routine entomological surveillance of dengue. Among them, BI remain as the most widely used larval index for vector management prior to and during epidemic incidences. Properly defined threshold values for BI are of essence, to assist the prediction of dengue epidemics and as a leading indicator for vector control. However, Sri Lanka still lacks effective thresholds for BI to initiate and drive dengue vector management activities. Hence, the present study attempts to define threshold values for BI based on an empirical modelling approach for the Kandy District of Sri Lanka.Item Eliminating female Aedes mosquitoes by spiking blood meals with toxicants as a sex separation method in the context of the sterile insect technique and Incompatible Insect Technique(Faculty of Graduate Studies, University of Kelaniya, Sri Lanka, 2016) Ranathunga, R.M.T.B.; Gunathilaka, P.A.D.H.N.; Udayanga, N.W.B.A.L.; Wijegunawardena, N.D.A.; Abeyewickreme, W.Ivermectin and spinosad have longevity reducing effects in several insect species, including disease transmitting mosquitoes after feeding blood contaminated with above toxicants on hosts. Therefore, this approach could play an enormous role in mosquito control operations by its use in the female elimination process during mass-rearing, enabling the release of only sterile males in the context of the sterile insect technique (SIT) and incompatible insect technique (IIT) with Wolbachia bacterium. Blood meals of cattle origin were spiked with diffeerenct concentrations (2, 4, 6, 8 and 10 ppm) of Ivermectin (Ivotec, 1% w/v) and Spinosad (Spinosyn, 12% w/v) and offered to a batch of adult Ae. aegypti (n = 300) and Ae. albopictus (n = 300) having 1:1 sex ratio in order to obtain an optimal dose for quick and total female elimination. After 24 hours all remaining females and males were aspirated out and transferred in to new cages separately. An additional blood meal with the initially fed concentrations of ivermectin and spinosad was provided after 24 hours, followed by a 48-hour observation of mortality. This experiment was repeated by feeding cattle blood containing a mixture of ivermectin and spinosad in 1:1 ratio from each concentration. Spiked blood with 8 ppm ivermectin and spinosad killed all the fed females of Ae aegypti and Ae. abopictus within 24 to 48 hours. It was observed that the number of fed females gradually increased, when decreasing the toxicity in combination of ivermectin and spinosad (1:1) in blood for both species. Male mortality did not show any significant differences during the study. Mixed feeding of blood with 8 ppm ivermectin and 8 ppm spinosad have shown as the viable treatment to eliminate female Ae aegypti and Ae. abopictus from laboratory colonies.Item Prevalence of dengue vector breeding habitats among selected Schools in Gampaha District of Sri Lanka(Faculty of Graduate Studies, University of Kelaniya, Sri Lanka, 2016) Radhika, N.M.L.; Gunathilaka, P.A.D.H.N.; Kasturiratne, K.T.A.A.; Abeyewickreme, W.Aedes aegypti and Aedes albopictus are the vectors responsible for the transmission of dengue viruses in Sri Lanka. Gampaha District has been ranked as the second most Dengue affected area. Recently, health authorities have identified that over 54 percent of schools have dengue mosquito breeding sites. However, the entomological information on the dengue vectors in School premises are not available. Therefore, the present study was carried out to identify the different breeding sites of dengue vector mosquitoes within school premises of randomly selected schools in Gampaha District of Sri Lanka. The premises of the selected schools (n= 34) were surveyed from May to October in 2016 for the availability of potential breeding habitats and the presence of vector breeding through entomological surveys. Collected vectors were identified morphologically and categorized inn to species level. All data on breeding habitats and number of vectors identified were recorded. According to survey, out of 34 schools surveyed; infested water-holding containers by Ae. albopictus were observed at 16 schools (47, 05%). Tree holes (Bridelia retusa tree), leaf axils of banana tree and fallen leaves were observed as natural breeding sites while, discarded plastic food containers plastic paint containers, broken pet bottles, tire marks on the ground, water retaining flower pots and plastic barrels were observed as artificial breeding sites. Breeding of Ae. aegypti was not identified among the school premises surveyed. Improper management of solid waste has affected positively in creating suitable habitats for vector breeding. Therefore, continuous monitoring of dengue vector breeding habitats and school based vector controlling interventions should be implemented through proper awareness and active participation for eliminating breeding habitats.Item Evaluation the Oviposition Behaviour of Aedes Aegypti and Ae. Albopictus Mosquitoes; Main and Secondary Vector for Dengue Transmission in Sri Lanka under Laboratory Conditions(Faculty of Graduate Studies, University of Kelaniya, Sri Lanka, 2016) Gunathilaka, P.A.D.H.N.; Ranathunga, R.M.T.B.; Udayanga, N.W.B.A.L.; Abeyewickreme, W.Studies on the oviposition behavior of Aedes albopictus and Ae. aegypti are critical for effective controlling of dengue vector breeding. However, less attention has been drawn on such aspects of vector biology and bionomics in Sri Lanka. Therefore, the objective of the current study was to evaluate the oviposition behavior of dengue vectors under laboratory conditions. Ovitraps with different sizes, colours, sources of water, sodium chloride (NaCl) concentration (0.2, 0.5, 1, 2 and 3%) and presence and absence of larvae were kept within separate cages of batches of 1000 Ae. albopictus and Ae. aegypti with (1:1, male: female ratio). Female mosquitoes were allowed to lay eggs and the number of eggs laid in different ovitraps were enumerated. ANOVA and cluster analysis were used to investigate the significance in the variations among oviposition behaviours. The mean number of eggs laid by both Ae. albopictus and Ae. aegypti increased with the increasing size of the ovitraps. Ae. albopictus indicated the highest mean number of eggs in 0.2% of NaCl than that of distilled water, while the egg laying preference reduced with the increasing salinity for both species. Drain water with low dissolved oxygen level (0.43 ± 0.12 mg/l) was the preferred water source for both species and indicated a significantly high oviposition rate in ovitraps with larvae. Black colour ovitraps attracted the majority of the gravid females, while white was least preferred. There were no significant variations among oviposition behaviors of Ae. albopictus and Ae. aegypti under laboratory conditions. Back coloured ovitraps were the preferred breeding place.Item Density of Aedes aegypti and Aedes Albopictus in some dengue endemic areas(University of Kelaniya, 2000) Hapugoda, G.P.G.M.D.; de Silva, N.R.; Abeyewickreme, W.; Rajamanthri, R.Item Aedes albopictus the “underrated” Asian Tiger(University of Kelaniya, 2010) Jayasooriya, D.H.S.W.; Gunawardene, Y.I.N.S.; Manamperi, A.; de Silva, H.J.; Abeyewickreme, W.Introduction The mosquito Aedes aegypti was thought to be the main vector responsible for virtually all dengue epidemics; while Aedes albopictus was considered a vector in which the virus is maintained but does not cause epidemics. Objective The study was conducted covering three endemic districts in Sri Lanka to determine the role of genus Aedes during dengue transmission. Methods and Material Mosquitoes were collected within a 350m radius from the location of the positive patients. Heads and abdomens of 63 pools were tested for DENV RNA with and RT-PCR-LH-(P32) assays Results Discussion Ae. albopictus was present in majority of the locations in all districts surveyed. Ae. albopictus was found in 13/17 (76.47%), 24/25 (96%)and 19/22 (86.36%) sites in Colombo, Gampaha and Kurunegala respectively. The RT-PCR-LH-(P32) assays indicated that 5/25 (20%) sites in Gampaha, 2/17 (11.76%) in Colombo and 6/22 (27.27%) in Kurunegala were positive for DENV. In Gampaha and Colombo there were 3 and 1 of DEN-2 positive pools respectively, while there were 2 and 1 of DEN-3 positive pools respectively. A higher number of positive pools (4/1or 21.05%) for DEN-1 and 1/1(5.26 %) for DEN-4 were found in Kurunegala. In Kurunegala one pool was positive for both DEN-2 and DEN-4 indicating the circulation of multiple serotypes within close proximity. Moreover one of the three DEN-2 positive pools in Gampaha consisting of only male Ae. albopictus mosquitoes is supportive of the belief of vertical transmission of DENV. In a DEN-4 positive location in Kurunegala HI was found to be10%, BI= 1and CI= 5.88 %while anotherDEN-2 positive site in Wattala showed HI of 5.55%and a BI of 5.55 suggesting active transmission. The abundance of Ae. albopictus in all districts and the findings indicating that100% of the positive pools were made of Ae. albopictus in this study highlights the importance of Ae. albopictus in the transmission dynamics dengue. The ability of Ae. albopictus to be infected with low viremia and the degree to which it permits replication within the mosquito itself could have an impact on the transmission and these verity of the disease. Co-circulation of two or more serotypes in a single pool or in different pools of mosquitoes within the same district is suggestive of hyper endemic transmission dengue in the three districts. The greater susceptibility of Ae. albopictus to infection by DENV is said to lead to greater virus adaptation. Sri Lanka as a whole would be at serious risks for multiple outbreaks in future. Our results indicate that Ae. albopictus is more efficient in dengue transmission than previously thought. The results shed light on the efficiency of Ae. albopictus as a vector in transmitting DENV in the absence or low abundance of Ae. aegypti in Sri Lanka. The present study suggests that Ae. albopictus sp is underrated in terms of transmission potential during peak transmission periods of dengue in Sri Lanka. Key words: RT-PCR-LH-(P32) RT-PCR-Liquid Hybridization with P32 radio isotope, HI-House hold Index, BI- Breteau Index, CI-Container Index,DENV-Dengue Virus Authors wish to acknowledge the financial assistance rendered by the NSF Sri Lanka (GrantNo:SIDA/2006/BT/02)and the IAEA (Grant NoTC SRL 6/028).