International Postgraduate Research Conference (IPRC)
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Item Susceptibility among Dengue Vectors to Malathion and Deltamethrin Insecticides in Three Selected Medical Officer of Health (MOH) Areas in the District of Gampaha, Sri Lanka(International Postgraduate Research Conference 2019, Faculty of Graduate Studies, University of Kelaniya, Sri Lanka, 2019) Dalpadado, C.P.R.D.; Amarasinghe, L.D.; Gunathilaka, P.A.D.H.N.Dengue has become the most important public health problem in Sri Lanka at present. As there is no effective vaccine or drug up to now, vector control is the main arm of disease control. Of these approaches, insecticide based control is used widely. However, excess use of chemicals and inappropriate application without determining the discriminative doses has given rise to the insecticide resistance among vector mosquitoes. Entomological surveys (Ovitap) were conducted in Negombo, Attanagalla and Dompe Medical Officer of Health (MOH) areas representing Urban, Sub urban and rural conditions, respectively from December 2016 December to June 2019. A total of 100 ovitraps each was placed outdoor and indoor in randomly followed by the collection after 5 - 7days. Eggs collected from ovitraps were reared under laboratory condition the laboratory (Temperature; 27 ± 10C: Relative Humidity 70 - 80%). The reared larvae were identified using standard taxonomic keys. Identification was further confirmed by identification of adults emerged from larvae . Adult mosquitoes were reared under optimized insectary conditions provided with sugar feeding. Batch of 140 mosquitoes from Aedes aegypti (Linnaeus) and Ae. albopictus (Skuse) was taken separately from the colony 24 hour after a sugar feeding. Each batch of mosquito was exposed to malathion (0.8%) and Deltamethrin (0.03%) insecticide susceptibility following the World Health Organization (WHO) defined procedures. The experiment setup was repeated for 2 times. Mortality and corrected morality was calculated using Abbortt’s formula after 24 hours’ exposure period. The results indicate that both dengue vectors have evolved high level of insecticide resistance to malathion (0.8%) and deltamethrin (0.03%) in the District of Gampaha except Ae. albopictus mosquitoes in rural areas. Ae. albopictus mosquitoes were found fully susceptible for 0.03% deltamethrin in Dompe (rural) MOH area and emergence of resistance (95.9%) was observed in Attanagalla (suburban) area. It was found resistant in Negombo area (83.8%) which is highly populated. Ae. albopictus mosquitoes were found fully resistant for 0.8% malathion in both urban (71.5%) and Sub urban (85.3%) areas while emergence of resistance was observed in Dompe MOH area (90.8%). Ae. aegypti was found resistance to both 0.03% deltamethrin and 0.8% malathion in Negombo area (88.1%, 53.5% respectively) and resistance was suggested for 0.03% deltamethrin in Attanagalla MOH area (91.8%) while it was found resistant for 0.8% malathion (89.8%) in the same area. Ae. aegypti was not recorded in Dompe MOH area during the study period. Since both the dengue vectors have developed resistance to WHO diagnostic discriminative doses for Malathion and deltamethrin intensity bioassays (five times and ten times of discriminative dose) are needed to be done to confirm the level of resistance in these vectors. Therefore, results highlight the requirement of continuous insecticide resistance monitoring incorporated with the surveillance and control efforts. Hence, use of evidence based discriminative doses of insecticide for each area should be vital to delay the development of resistance among natural populationItem Assessment of Possible Risk Factors Affecting Transmission of Dengue in the District of Gampaha Based on Reported Dengue Cases(International Postgraduate Research Conference 2019, Faculty of Graduate Studies, University of Kelaniya, Sri Lanka, 2019) Perera, E.H.L.; Viswakula, S.; Gunawardene, Y.I.N.S.; Subasinghe, U.; Hapugoda, M.D.Dengue is a fast spreading arboviral infection transmitted by the bite of infected females of Aedes aegypti (Linnaeus) and Ae. albopictus (Skuse). According to the Epidemiology Unit, the second highest number of dengue cases is reported in the District of Gampaha, Sri Lanka over past ten years. Objective of this study was to investigate the entomological and socio-economic risk factors affecting transmission of dengue in laboratory-confirmed dengue case reported stations in the District of Gampaha. Laboratory confirmed positive dengue patients (n=100) by dengue NS1 antigen test during the period of June, 2018-August, 2019 were selected. Entomological surveillance was conducted by visiting to each patient within one week of notification of a positive case. For the collection of socio-economic data, an interviewer-administrated questionnaire was used. Adult Aedes mosquito samples collected using a back-pack aspirator showed, 98.64% (73/74) of Ae. albopictus and 1.35% (1/74) of Ae. aegypti mosquitoes. Larval collection using standard larval surveillance techniques showed 92.96% (185/199) and 7.04% (14/199) of Ae. albopictus and Ae.aegypti larvae respectively. The highest House Index (55.17%-16/29), Container Index (28.89%-13/45) and Breteau Index (44.83%-13/29) were reported in the month of June, 2019. The major Aedes breeding place was identified as plastic buckets/barrels (48.6%-84/173) that being used to discard waste. Piped borne water (88%-88/100) was the major water source of the house-holds. Water source of tube well (9%-9/100) was the next popular water source and 66.67%(6/9) of tube wells were positive breeding places for Aedes larvae. Average homestead of the premises of dengue patients was 16.14 perches. From the 100 dengue cases, 67 cases were from middle of town areas, while 2 were from rural areas. Vegetation coverage of the 78% (78/100) house-holds were grass, bushes and small trees and 3% (3/100) house-holds didn’t have any vegetation coverage. The major mosquito prevention method was usage of mosquito nets (54%-54/100) and among dengue patients 7% (7/100) of dengue patients weren’t using any mosquito prevention method. High density of Ae. albopictus mosquitoes, was reported although Ae. aegypti is the major vector of dengue. Therefore, it is required to draw more attention about the Ae. albopictus breeding sites in dengue control programmes. Participants from the study sites were well aware about the disease but still there is a lack of knowledge on breeding sites and vector control methods. Drawbacks in the waste disposal methods, lack of cleanliness in gardens, unplanned water sources and neglecting preventive actions could be considered as the possible risk factors.Item Antagonist Potential of Vorticella microstoma on the larval development of Culex gelidus and Aedes aegypti(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Ranasinghe, H.A.K.; Amarasinghe, L.D.Selection of effective biological agents which can suppress the growth of immature stages of mosquito vectors has drawn a wide attention in disease control programmes. Vorticella microstoma is a parasitic ciliate which has an ability to parsitize on larval stages of mosquitoes and inhibit their growth. However, the potential antagonist activity has not been evaluated in detail in Sri Lanka. Fifteen each of first, second, and third instar larvae of Cx. gelidus and Ae. aegypti were placed in separate V. microstoma culture bottles at room temperature (27± 2 ºC) and mortality rate of mosquito larvae was observed after 48 h. Bioassay was done in completely randomized design with controls. The first, second, and third instar larvae of Cx. gelidus were allowed to be infested with the trophont stage of V. microstoma. Heavily infested larvae with V. microstoma were identified by the presence of parasites attached all over the body surface. V. microstoma trophonts did not usually attach to siphon region of mosquito larvae when they are alive. None of the instar levels of Ae. aegypti showed susceptible to infection with V. microstoma. In the first instar larvae of Cx. gelidus, thorax and the abdominal segments had the attachment of 30–35 trophonts of V. microstoma, whereas 2–4 trophonts had attached to the anal papillae. Higher densities of V. microstoma were found in abdominal segments of second instar larvae of Cx. gelidus, whereas total of 45–50 trophonts had attached to thoracic region and abdominal segments. Fifty to fifty-five trophonts of V. microstoma had attached to third instar larvae of Cx. gelidus, with higher densities in the anal papillae followed by abdominal segments; in the anal papillae, 20–25 trophonts were found to be attached. V. microstoma infection was positively correlated with the body size of mosquito larvae. The percentage mortality of the first, second, and third instar larvae of Cx. gelidus did not significantly differ from each other (One-Way ANOVA: IBM SPSS Statistic Software, P<0.05). Therefore, the study results would be a new avenue to work on environmentally agreeable manner in reducing the Cx. gelidus vector mosquito populations.Item Isolation of a Potential Microbial Agent for Controlling Dengue Vector Mosquitoes in Sri Lanka(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Induwara, R.; Fernando, M.; Ranathunge, T.; Parakrama, G.; Hapugoda, M.Controlling dengue vector mosquito is the most appropriate controlling method for dengue in the absence of an effective drug or a vaccine for dengue viruses. Vector control can be performed using variety of approaches such as environmental management, chemical and biological control etc.; which have been used in recent vector control activities. Due to the development of resistant varieties against chemical insecticides, the present study attempted to identify larvicidal activity of bacteria collected from the natural environment, as an ideal environmental friendly and effective strategy for controlling Aedesaegypti (Linnaeus), a dengue vector mosquito species. Zero dengue infection was reported in Pudumurippu area in the District of Kilinochchi, Northern Province of Sri Lanka in a preliminary survey. Spore forming microbes were isolated through spread plate technique using water and sediment samples collected from a reservoir in Pudumurippu. Larvicidal activity of each isolate was tested by Ae. aegypti third instar Larvae (L3) in vitro. The highest larvicidal activity was observed in an isolated bacterium from a reservoir water sample under laboratory and field conditions. This bacterium was presumptively identified and subjected to 16s-rRNA sequence analysis. Larvicidal activity of this bacterium was compared with a currently used Bacillus thuringiensisisraelensis (Bti). As well as optimum physiological characteristic features of isolated strain was determined by growing the bacteria strain on nutrient agar supplemented with different NaCl concentrations and different pH values. Isolated bacterium was confirmed as a new strain of Bacillus cereus (SL001; MG827268). This bacterial strain showed the highest larvicidal activity at 5% (1×105 CFU/ml), with mean cumulative mortality rate 92±4.1% and 84.2 ±5.3% at 48 hours’ post challenged under laboratory and field conditions respectively. When compared with Bti, this novel strain showed significantly higher (p < 0.05) larvicidal activity. B. cereus (SL001) displayed high growth rate while tolerating wide range of salinity (0-30gL-1) and pH (6-10). Based on the findings, B. cereus (SL001) with the highest larvicide efficiencies could be an ideal candidate for biological controlling of Ae. aegypti dengue vector mosquitoes in Sri Lanka. Further analysis of this bacterium is on going at present.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.