International Postgraduate Research Conference (IPRC)
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Item 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 Detection of Dengue Viral Migration to Sri Lanka(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Withanage, G.P.; Hapuarachchi, H.C.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.Dengue is one of the most important mosquito-borne viral infectionsin Sri Lanka.The causative agent is Dengue Viruses (DENV) and the primary vector of the virus is Aedesaegypti(Linnaeus) while Ae. albopictus (Skuse) is the subsidiary vector. The current research was focused on the detection of DENV serotypes and genotypes circulating in mosquitoes during the dengue epidemic in June and July, 2017 in the EriyawetiyaGramaNiladhari division, where one of the dengue high-risk area in Kelaniya Medical Officer of Health (MOH) area in the District of Gampaha, Sri Lanka. Aedesmosquitoes were collected following WHO guidelinesandthe field-caught mosquitoes were transported to the laboratory for species identification and subsequent analysis. Head and thorax of each mosquito was removed and mosquito samples were pooled separately. Total RNA was extracted from mosquito samples and semi-nested Polymerase Chain Reaction (PCR) was performed to identify DENV serotypes present in the mosquito samples. The results of the PCR indicated the presence of DENV2 in both Ae. aegypti (1/5) and Ae. albopictus (1/27) mosquitoes. Then complete Envelope (E) gene was amplified with DENV2 specific primers for genotyping of the virus which is required to identify the molecular evolution of the DENV2. Prior to sequencing the PCR products were purified and sequencing results were analyzed usingLaserGene software. The generated sequences were aligned with retrieved DENV2 sequences available at NCBI database and the phylogenetic trees were developed using MEGA7 software with General Time Reversible (GTR) substitution model with gamma distributed rates. The robustness of clades was determined by using bootstrap analysis of 500 replicates. The result of the phylogenetic analysis illustrates that the E gene sequences of DENV2 obtained from two DENVpositive mosquito poolsbelong to DENV2 Cosmopolitan Clade Ib, which has been the dominant strain in South-East Asia, specially Singapore, Indonesia, Malaysia, and China since August, 2015.The evidence suggests recent introduction of this DENV strain into Sri LankaItem Larvivorous Potential of Four Fish Species Against Dengue Vectors in The Kandy District of Sri Lanka(Faculty of Graduate Studies, University of Kelaniya, 2015) Kusumawathie, P.H.D.; Jayasooriya, G.A.J.S.K.; Gunathilaka, P.A.D.H.N.; Hapugoda, M.D.; Abeyewickreme, W.There is a growing concern on the effects of insecticides that are used for the control of mosquito vectors of human diseases. Introducing a self propagating predator into the vector breeding habitats may present continual biological control of vector population. In order to control dengue vectors, namely, Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) (Diptera: Culicidae), larval control is one of the major strategies. The objective of this study was to evaluate the larvivorous potential of fish species against dengue vectors. Four fish species, namely, Poecilia reticulata (guppy), Aplocheilus dayi (Udahandaya), Oreochromis niloticus (Tilapia) and Puntius bimaculatus were collected from natural breeding habitats. They were placed in fish stock cement tanks at the Regional Office of the Anti Malaria Campaign in Kandy till they were used for the study that was conducted from January – June 2015. In order to determine the larvivorous potential of the fish species, five fish of each species were placed in separate enamel trays containing de-chlorinated water and allowed to acclimatize for half an hour. Afterwards, a batch of 25 Ae. aegypti larvae was introduced to each tray and counted the number of larvae remaining after 5, 10, 30 minutes, 1 and 24 hours. When all 25 larvae are consumed, another batch of 25 larvae was introduced to those trays and this was continued till the study was completed. At the end of study period (24 hours), the number of larvae consumed per fish for each fish species was determined. This study was carried out in the laboratory with 6 replicates. The experiment was repeated for Ae. albopictus. Overall, Oreochromis niloticus, Poecilia reticulata, Aplocheilus dayi and Puntius bimaculatus consumed 97.6, 66.9, 49.3 and 47.5 Ae. aegypti and 98.4, 62.3, 51.2 and 45.4 Ae. albopictus larvae respectively. All four fish species showed larvivorous activity and Oreochromis niloticus showed the highest larvivorous potential followed by Poecilia reticulata. Since Ae. aegypti and Ae. albopictus breed in water storage containers such as cement tanks, barrels and ornamental ponds, the fish species that showed high larvivorous potential may be considered for dengue vector larval control in such containers. In order to achieve good community participation for the use of larvivorous fish for dengue vector control, effective health education is of utmost importance.Item IGamma- ray irradiation as a method for sterilization of males of Aedes albopictus (skuse) for its control through Sterile Insect Technique (SIT)(Faculty of Graduate Studies, University of Kelaniya, 2015) Harishchandra, J.; Abeyewickreme, W.; Hapugoda, M.D.; Premaratne, R.G.; Gilles, J.R.L.Sterile Insect Technique (SIT) can be integrated with other conventional control methods to suppress population of Aedes albopictus, a vector of dengue in Sri Lanka. A study was conducted to determine the effective dose of gamma radiation for producing sterile males of Ae. albopictus mosquitoes. A batch of male pupae (n=32) aged 24-48 hours in F1 was irradiated using Gamma 220, (Co60) irradiator with 25, 30, 40, 50, 60 and 70Gy in duplicates. Adult emergence rate and male longevity were recorded. Virgin females from the same cohort (F1) were introduced into each cage for mating and fed blood starting 5 days after emergence. Then, females were isolated in individual tubes and hatching rate of individual egg batches was calculated after two weeks of egg maturation in hatching solution. Spermacheca of female mosquitoes were dissected and insemination rates were calculated after egg laying. Male mosquito pupae in F1 showed low mortality (0-3.12%) immediately after exposing to radiation (0-48 hours). After 14 days of the irradiation, survival rate of male mosquitoes in F1 were 82.7%, 79.7%, 56.3%, 73.4%, 76.6%, 76.1% and 56.3% when irradiated at 0Gy, 25Gy, 30Gy, 40Gy, 50Gy, 60Gy and 70Gy respectively. No significant difference was observed among survival of males in different doses except in 30 Gy and 70 Gy (Kaplan-Meier survival analysis, Log Rank test). Low survival rate at 30 Gy could be due to mosquito handling errors and need to be retested. Based on the hatching rate of the F2 progeny, males (F1) showed 89%, 93%, 97%, 99%, 99% and 100% sterility when irradiated at 25Gy, 30Gy, 40Gy, 50Gy, 60Gy and 70Gy respectively. Insemination rate of the irradiated males among different doses and controls were above 90% in F1 based on spermatheca dissection of bloodfed females (F1). 50 Gy is recommended as the most suitable radiation dose to produce 99% sterility in Ae. albopictus males which has 76% survival after 14 days of irradiation. This dose can be used to produce sterile males of Ae. albopictus for a SIT programme.Item Characterization of Anopheline larval habitats and species composition of aquatic macro-invertebrates in Trincomalee District, Sri Lanka(Faculty of Graduate Studies, University of Kelaniya, 2015) Ranathunga, R.M.T.B.; Gunathilaka, P.A.D.H.N.; Kannangara, D.N.; Abeyewickreme, W.; Hapugoda, M.D.Malaria control methods that aim to reduce adult vector populations by targeting their aquatic immature stages. A better fundamental understanding of the biology and ecology of these essential stages could contribute to the implementation of current control methods and to the development of novel strategies. Objective of this study was to examine breeding habitat diversity and analysis of richness, diversity and geographical distribution of Anopheline larvae and the species composition of aquatic macro-invertebrates in their oviposition sites in Trincomalee District. Fifteen major permanent breeding places in five possible malaria sensitive sites (Gomarankadawala, Ichchallampaththu, Mollipothana, Padavisiripura and Thoppur) in Trincomalee District were selected. Anopheles larvae and macro-invertebrates were collected using standard methods for 16 months (April, 2013-July, 2014) and they were identified microscopically. The Shannon diversity index (H') was used to characterize species diversity at the five study sites by its abundance and evenness of the species present. ANOVA were used to analyze the correlation between macro-invertebrates and mosquito larval abundance. In total, 4478 including 11 species of Anopheles larvae were identified. An. subpictus, An. nigerrimus and An. peditaeniatus (71%) were the most abundant and widely-distributed species. Anopheline larval diversity was highest in Mollipothana (H‘=1.986). Whereas in Gomarankadawala, Ichchallampaththu and Thoppur where H‘=1.721, H‘= 0.857 and H‘=0.762 respectively. In total, 28 species of aquatic macro-invertibrates were identified and highest diversity was recorded in Mollipothana, Gomarankadawala and Ichchallampaththu (H‘=3.14-2.56). This deduces that the species richness and diversity of Anopheles mosquitoes and macro-invertebrates are higher in these areas. The presence of permanent breeding places may be the factors for this phenomenon. This study represents the first systematic update to the distribution of macro-invertebrates associated with Anopheles mosquito oviposition sites in Trincomalee District. Knowledge generated on the ecology of Anopheles mosquitoes will help to eliminate malaria vectors in the country.Item Assessment of possible social and entomological risk factors affecting transmission of dengue in the District of Gampaha(Faculty of Graduate Studies, University of Kelaniya, 2015) Withanage, G.P.W.K.; Abeyewickreme, W.; Hapugoda, M.D.Dengue is the most important mosquito-borne viral infection transmitted to humans in Sri Lanka causing more than 30,000 cases annually. The objective of the study was to identify possible social and entomological risk factors affecting transmission of dengue in Gampaha district where the second highest number of dengue cases recorded during last ten years. Four Medical Officer of Health (MOH) areas were selected based on annual number of dengue cases greater than 250 for last ten years. One GramaNiladhari (GN) division was selected from each MOH area as a study area where the highest number of dengue incidents reported. In each study area, a cluster of 150 households was selected and household and entomological surveys were conducted. The selected areas were Eriyawatiya (Kelaniya MOH), Welikadamulla (Wattala MOH), Akbar town (Mahara MOH), and 3-Kurana (Negombo MOH) GN divisions. There were 2577 population in 600 households in the study areas and 53.5% were females. Average size of a homestead was 17 perches and most of the households (98%-588/600) were individual house. Main vegetations in the homesteads were grass and bushes (97%-583/600) and potential breeding places of dengue vector mosquitoes were observed in 96.1% (577/600) households. Main dengue vector mosquito present in the captured Aedes adult mosquitoes was Aedes albopictusis (92.9%-183/197). Most prominent breeding places were discarded bottles and tins (4.0%-15/371), plant axils (9.7%-36/371), plastic containers (26.4%-98/371), and roof gutters (4.3%-16/371) in the observed premises and 9.2% (34/371) were Aedes larvae positive. Most frequent mosquito preventive measure was bed-nets (30.3%-182/600), but mosquito coils (30.8%-185/600) and vaporizers (17.6%- 106/600) were frequently used. Participants have adequate knowledge about the disease, but they neglect preventive measures and highly depend on government vector control programs. Unplanned households, crowded conditions, poor waste management systems, and negligence to pursue preventive measures may be possible risk factors and therefore, frequent encouragement is needed to control dengue transmission.