Conference Papers

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This collection contains abstracts of conference papers, presented at local and international conferences by the staff of the Faculty of Medicine

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Establishment and maintenance of laboratory colonies of Aedes albopictus mosquitoes
(University of Peradeniya, 2015) Wijegunawardana, A.D.; Gunathilaka, H.N.; Dassanayake, R.; Gunawardene, Y.I.N.S.; Abeyewickreme, W.
With a mission of "providing authenticated, high-quality Aedes albopictus mosquito rearing information to the research community" maintenance of a Ae. albopictus mosquito colony was started. All environmental facilities inside the insectary were carefully maintained to better suit the Ae. albopictus mosquito colonization. The mean temperature of 27°C (± 0.5°C) was constantly maintained inside the insectary. Wet towels on adult mosquito cage racks were used for proper maintenance of humidity. Lighting was using fluorescent light and regulated with 16:8 hour continuous dark and light period. Pest insect was controlled to ensure essential absence of ants and cockroaches. This was achieved without any harm to the mosquito colonies either directly or by contamination with toxicants transported by pests. An adult mosquito trap placed inside the insectary was used to monitor released mosquitoes. Consistent effort was also made to improve the level of cleanliness inside the insectary. Written guidelines were given to each person responsible for a task. Insectary operations included egg counting, preparation of hatching bottles with boiled distilled water following cooling to room temperature, egg hatching, larvae rearing with International Atomic Energy Agency (IAEA) recommended diet of tuna meal, bovine liver powder, brewery yeast and vitamin complex in a ratio of 37.5:27:10.5:2 g in 1L up to one week, pupae counting and putting into adult emergency cages, adult male feeding with 10% sugar solution with Vitamin complex, adult female blood feeding from 4th day onwards with bovine blood, placing egg laying cups and collecting egg laying cups, drying egg papers and starting next generation from the dried eggs. Adult mosquito cages were blood fed every 4th day after emergence from pupa and for quality control reasons each adult cage was blood fed only 3 times and there after only 10% sugar solution with vitamin syrup was supplemented until all adult mosquitoes died. Documentation for maintenance and data record was maintained and updated daily. Records included larvae feeding records, larvae tray maintenance and cleaning charts, adult feeding records with both sugar solution and blood, insectary cleaning records with time and dates. Number of eggs and percentage of egg hatching, larvae death, pupation, adult emergence, egg laying and adult mosquito death with respect of the sex and time difference were recorded. For bio-safety reasons all discarded material from larvae trays, egg laying cups and adult cages were boiled thoroughly to facilitate total destruction of the contaminated mosquito eggs. All other infectious material were incinerated. Finally, all above conditions facilitated achievement of 100% egg hatching rate within maximum of 24 hours, 100% survival of larvae to pupa (~ 7 days), 100% survival of pupated larvae to adult emergence (~ 2 days) and 95.5% adult survival up to 12 days. No difference was observed on adult longevity between males and females within first 12 days of adult emergence. However, approximate life span for males (-17 days) was lower than the females (~ 25 days) and the mortality was regular through all generations (Fl to F21).
Male isolation of dengue vector Aedes mosquitoes for sterile insect technique by spiking blood with ivermectin
(Sri Lanka Association for the Advancement of Science, 2015) Abeyewickreme, W.; Wijegunawardana, N.D.A.D.; Ranathunge, R.M.T.B.
There is an increasing demand for exploration of the potential for applying the Sterile Insect Technique (SIT) in area-wide integrated vector management (AW-IVM) in many countries. However, because female mosquitoes, unlike male mosquitoes, can transmit disease, means to eliminate them from the mass production process are a critical pre-requisite. In addition, the efficiency of the SIT programme could be increased by not releasing female mosquitoes due to the fact that sterile males can then only focus on wild females to achieve mating. Therefore, mosquito SIT programmes success will depend on exclusive release of sterile males, which is impossible on a large scale without efficient sex separation methods. Many different sex separations for different stages of mosquitoes are currently being attempted to successfully establish a sexing mechanism for the above purpose. Among them, the mostly attempted methods broadly include genetic sexing methods, molecular methods, mechanical methods and behavioral methods. For all blood feeding mosquitoes, sex separation could occur at the adult stage by spiking blood with insecticides (malathion, dieldrin) or other mosquito toxins (ivermectin, spinosad), as behavioral tools. In this study, a veterinary preparation of ivermectin (Ivotec,1% w/v) which is used to treat dogs infested with the filarial worm Dirofilaria repens in Sri Lanka was used as a tool for sex separation of adult Aedes albopictus and Aedes aegypti mosquitoes by feeding on spiked blood. In an initial attempt by spiking blood with 5 parts per million (5 ppm) ivermectin, 40% of the blood–fed females could survive beyond 24 hours post feeding. However, when the dosage of ivermectin was increased to 7 ppm the survival rate decreased below 40%. Since there had been high mortality of both females and males even in the control cages in which mosquitoes were blood-fed without ivermectin it was suggested that the age of the mosquitoes used may be a factor for high mortality of mosquitoes in the control experiment. When the experiment was repeated with 7 ppm ivermectin in blood using 4-5 day old mosquitoes more than 65% mortality could be observed within 12 hours post feeding in the experimental cage with a significant difference in mortality compared to the control
Entomological investigations on malaria vector studies in earlier conflict areas of Sri lanka after 30 years
(University of Peradeniya, 2014) Gunathilaka, P.A.D.H.N.; Fernando, M.A.S.T.; Hapugoda, M.D.; Wijeyerathne, P.; Wickremasinghe, A.R.; Abeyewickreme, W.
Entomological investigations on the abundance of malaria vector mosquitoes have not been studied in northern and eastern parts of the Sri Lanka over the past 30 years due to the separatist war. The main aim of this study was to explore diversity and abundance of Anopheles mosquitoes in earlier conflict areas in Sri Lanka. Monthly entomological monitoring was carried out at 60 possible malaria sensitive localities situated approximately 12 km apart in 15 selected sentinel sites in Ampara (4), Batticaloa (3), Mannar (3) and Trincomalee (5) districts for 32 months (June 2010 to August 2013). Adult mosquitoes were collected by WHO recommended techniques. Out of 701,356 anophelines collected, An. culicifacies was noted only in Ampara, Batticaloa and Trincomalee Districts. Although the main vector An. culicifacies (n= 1,876) was low in numbers, the presence of secondary vectors including An. subpictus (n= 205,594) were high in these areas. An. nigerrimus (n= 227,057), An. barbirostris (n= 35,150), An. vagus (n= 21,161), An. pallidus (n= 17,403), An. annularis (n= 4,882), An. varuna (n= 3151), An. tessellatus (n= 718) and An. aconitus (n= 591) were the other species reported. There was a change in breeding habitats of An. culicifacies and An. subpictus. They were found more conducive to breeding in built wells, brackish water habitats and waste water collections which were below 3 mg/l of dissolved oxygen (2.85 ± 0.03). These results indicate that particularly An. culicifacies has adapted to breed in wide range of water bodies including waste water collections although they were earlier considered to breed in clean and clear water with high dissolved oxygen. The adaptation of the major and subsidiary vector mosquitoes to widespread water bodies (along with increase in imported cases) could be a potential factor for the increase in the incidence of malaria in the future even though reported cases are low at present. Further, entomological surveillance detected the presence of An. jeyporiensis from the country after 106 years. Hence, more classical entomological studies are required to describe species currently found in the country; revision of morphological identification keys is a step in this direction. Financial assistance given by the Global Fund for Aids, Tuberculosis and Malaria (GFATM-Round 8-SRL809G11M.) through TEDHA malaria elimination program is acknowledged.
Dengue vector surveillance in a dengue hot-spot in Sri Lanka
(Faculty of Tropical Medicine, Mahidol University, 2007) Sumanadasa, S.D.M.; Hapugoda, M.D.; Perera, D.; Bandara, S.; Mansoor, M.A.; Peris, I.; Abeyewickreme, W.
BACKGROUND: In South Asia, dengue has been declared as one of the most, fast-spreading vector-borne diseases. Therefore, mosquito surveillance is important for early detection of outbreaks along with implementation of prompt control activities. OBJECTIVES: To identify entomological risk factors with regard to transmission of dengue in a dengue hot-spot. Seventy five human dwellings in Vehara in the Kurunegala District of the Western Province were selected based on high disease incidence during 2000-2004, high Aedes as well as human population density and increased building activities. Entomological surveillance was done during May-August, 2007. RESULTS: The house Index ranged from 2.67% to 5.33% for Aedes aegypti while it for Aedes albopictus was 1.33% to 6.60%. The container index ranged from 23.67% to 29.33% for Ae. aegypti and from 1.33% to 18% for Ae. aibopictus. Man biting rates of 0.43-5.78 bites/man/hour were estimated for Ae, aegypti, while it ranged between 0.49 and 1.33 for Ae. aibopictus. The most common breeding place for Aedes species was plastic baskets (16%, n=12). DISCUSSIONS: Vector surveillance showed that the predominant vector species present in the study area was Ae. ageypti. Aedes mosquito larval densities and adult biting rates were sufficient to promote outbreaks of dengue in this study area. Community must be educated regarding effective measures to protect them from dengue. Their cooperation should be elicited in the early detection and elimination of vector species by source reduction, environmental management and personal protection measures.
Potential use of IGR Pyriproxifen for control of dengue vector Aedes albopictus
(Sri Lanka College of Microbiologists, 1999) Gunawardene, Y.I.N.S.; de Silva, N.R.; Abeyewickreme, W.
Dengue Fever (DF) Dengue Haernorrhagic Fever (DHF) is now a significant problem in Sri Lanka. The incidence of DHF has increased cyclically since the first recognized outbreak in 1989. Of the 203 suspected DHF cases recorded in 1989, 87 were confirmed and 26 deaths were reported. By 1997, 5882 clinical cases, 1558 serologically confirmed cases and over 300 deaths have been reported. Without an effective vaccine against dengue, and considering the clinical difficulty in managing DHF cases, vector control has become an important com¬ponent in the integrated attempts for dengue con¬trol. Aedes aegypti and Aedes albopictus not only transmit dengue but are also a nuisance and cause annoyance by their day biting behaviour. In this study an attempt was made to control Ae. albopictus by exploiting its oviposition behaviour, us¬ing an Insect Growth Regulator (IGR), Pyriproxifen. The IGR is known to interfere with the synthesis and deposition of chitin in insects and thereby prevent growth and development. Different concentrations of the IGR (0.01 g/1 -0.08g/1) were tested against Ae.albopictus, in laboratory experiments. The most effective larvisidal concentration of Pyriproxifen for Ae.albopictus was determined to be 0.03g/1. Field trials were also carried out to determine the feasi¬bility of employing used automobile tyres contain¬ing Pyriproxifen as ovitraps against Ae.albopictus. Tyres containing water were treated with Pyriproxifen (0.03g/1) and the emergence of adults was recorded. Pyriproxifen at a concentration of 0.03g/1 was found to inhibit the emergence of adult Aedes mosquitoes up to 6 weeks.
Silent transmission as a risk factor affecting transmission of dengue fever.
(Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, 2007) Hapangama, H.A.D.C.; Hapugoda, M.D.; Gunawardene, Y.I.N.S.; Premaratna, R.; Dayanath, M.Y.D.; Abeyewickreme, W.
The global incidence of dengue fever has increased by more than four-folds over the last 30 years, making it the most threatening mosquito-borne viral disease at present. Objective of this study is to determine the role of silent transmission •on' incidence of dengue. A total of 40 households, living within a 300 m radius of seven selected ‘confirmed dengue cases at different dengue high risk localities in Gampaha District were recruited for this study. A minimum of three inhabitants were tested for anti-dengue antibodies using a commercial kit to determine the prevalence of silent transmission on dengue infection in each households. Entomological surveillance was carried out in all seven localities. Out of 40 households, 26 (65%) houses were positive for dengue viral infection. Total of 148 inhabitants (68 Males); mean age; 35.9 years were enrolled. Of the 148, 41 (27.7%) had evidence of exposure to dengue virus [positive for IgM: 28/41(68.4%), IgM'& IgG: 7/41(17%) and IgG: 6/41(14.6%)]. Out of 28 primary infections, 20(71.4%) were asymptomatic. Of the 7 secondary infections, 1(14.28%) was asymptomatic. Of the 6 previous exposures to dengue 4(66.67%) were asymptomatic. Of the 7 localities investigated, 1 had >50%, 4 had >25% and 2 had <25% clustering of cases. Ae. albopictus found in all seven localities and Ae. aegypti found only in two localities. This study suggests presence of silent transmission of dengue virus with a trend towards clustering around cases and also this shows that the presence of vectors increases the incidence of dengue.
Ovi-trap based surveillance for Aedes in dengue endemic Gampaha District, Sri Lanka during rainy season
(Faculty of Tropical Medicine, Mahidol University, 2008) Hapangama, H.A.D.C.; Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Benjamin, S.; Abeyewickreme, W.
Dengue is endemic in Gampaha District, Sri Lanka. Aedes aegypti and Aedes albopictus have been incriminated as vectors in urban and suburban environments. An ovitrap-based weekly surveillance was carried out to determine the distribution and abundance of Dengue vectors, in six dengue endemic areas; Galudupita, Gongitota, Wanawasala, Kerangapokuna, Mabole and Welikadamulla in the Gampaha District during May and June 2008 in both indoors and outdoors. Galudupita and Gongitota gave the highest ovitrap indices, both indoors and outdoors as 97% and 73% respectively. Of the total positive ovitraps(1320), positivity for Ae. albopictus, Ae. aegypti and both were 63.5%, 32.9% and 3.6% respectively. Highest mean number of Ae. aegypti and Ae. albopictus larvae per ovitrap in both indoor and outdoor ovitraps were observed Galudupita and Gongitota respectively. However, statistical analysis showed that ovitrap index of Ae. aegypti in six study was not significantly different from each other (P > 0.05). Outdoor mean number of Ae. aegypti larvae per ovitrap in Galudupita, was significantly higher than other sites (F=3.587, P=0.012). Indoor Ae. albopictus ovitrap index of Gongitota was significantly higher than of Welikadamula (F=2.59, P=0.046) while outdoor Ae. albopictus ovitrap index of Gongitota and Wanawasala was significantly higher than of Welikadamula F=3.19 P=.0.02). Results irnplicate that Ae. albopictus prefers more vegetation as observed in Gongitota and Wanawasala. In highly populated and urbanized Galudupita, area with less vegetation, Ae. albopictus population was less than Ae. aegypti even outdoors. Study implicates that Ae. albopictus is the predominant indoor and outdoor breeder during rainy season.
GIS mapping of Lymphatic Filariasis endemic areas in Gampaha district, Sri Lanka; based on the epidemiological and entomological screening
(Faculty of Tropical Medicine, Mahidol University, 2009) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Manamperi, A.; Abeyewickreme, W.
BACKGROUND: The health issues related to vector borne diseases appear always to be related to space and time. Therefore it is ideal to link Geographical Information Systems (GIS) with epidemiological and entomological data to monitor spread of infection and target control strategies. OBJECTIVE: The objective of this study was to develop a site directed GIS map for lymphatic filariasis (Lf) dispersed areas in Gampaha district, Sri Lanka as a guide to target control activities. METHODOLOGY: Epidemiological and entomological screening of Lf was done in nine pre-identified endemic areas in Gampaha district, using night blood screening and pool-screening PCR-ELISA protocols respectively. RESULTS: Overall, 1073 (286 children, 787 adults) from 9 sites were examined. Mf-positive cases were detected in 2 sites, with a prevalence rate of 10.5% (Hekiththa) and 3.4% (Peliyagoda) with over 30% Mf prevalence in adult mosquito populations. The overall prevalence of mosquitoes with L1-L2 larvae of W. bancrofti ranged from 0%-8.54% by dissection and point estimates of infection prevalence, as assayed by PCR-ELISA, ranged from 0% - 35.4%. According to geographical data, the highest number of cases was found at altitudes between 2.5-3.5 m and highly populated areas where transmission appears to be taken place. Questionnaires indicated limited community awareness can be a reason for the fairly static infection rate prevalent in Peliyagoda sentinel site. DISCUSSION: The maps derived indicate the substantial extent as well as the marked variability in the geographical distribution of Lf in Gampaha, demonstrating site related trends.
Surveillance for dengue vector mosquito in Kurunegala district, Sri Lanka
(Elsevier, 2008) Sumanadasa, S.D.M.; Hapugoda, M.D.; Peiris, I.; Perera, D.; Bandara, S.; Mansoor, M.A.C.M.; Abeyewickreme, W.
BACKGROUND: Dengue vector surveillance is an important tool to determine the time and area/s to initiate control action. Our objective was to identify entomological risk factors with regard to transmission of dengue in a dengue hot-spot. METHODS: In the study 75 human dwellings in a dengue hot-spot in the District of Kurunegala was selected based on high disease incidence during 2000—2004, high Aedes as well as human population density and increased building activities. House to house mosquito surveillance was carried out from 08.00 am to 12.00 noon during May—August, 2007. Larvae and adult Aedes mosquitoes were collected indoors and outdoors using normal larval surveillance and human landing diurnal collection techniques respectively. Environmental and sociological data were obtained from households by interviewer administered questionnaires and observations. RESULTS: The house index for Aedes aegypti and Aedes albopictus ranged from 1.33%—6.60% and 2.67%—5.33% respectively. The Breatu index ranged between 23.67%—29.33% for Ae. aegypti and 1.33%—18% for Aedes albopictus. Man biting rates of 0.43—5.78 bites/man/hour were estimated for Ae. aegypti, while it ranged from 0.49—1.33 for Ae. albopictus. The most common breeding place for Aedes species was plastic baskets (16%, n = 12). More than half of the households (61%, n = 45) stored water in large cement tanks for their daily activities. Majority of the group (99%, n = 74) had a basic knowledge on dengue mosquito breeding places (97%, n = 73), their life cycle (95%, n = 71) and mode of transmission (99%, n = 74). Domestic waste of majority of households was carried away daily by the local authorities (n = 69). CONCLUSION: These results suggest that a vector control program should be adopted to reduce Aedes population levels below dengue transmission thresholds. Discarded containers of various types were identified as potential mosquitoes breeding habitats. Therefore, community must be educated on effective vector control measures to contain the transmission levels. © 2008 Elsevier Inc.
Species succession of malaria vector mosquitoes and physicochemical factors affecting their abundance in rice ecosystems in Trincomalee, Sri Lanka
(University of Kelaniya, 2013) Gunathilaka, P.A.D.H.N.; Ranathunge, R.M.T.B.; Fernando, M.A.S.T.; Hapugoda, M.D.; Wijeyerathne, P.; Wickremasinghe, A.R.; Abeyewickreme, W.
Background: Mosquito-borne diseases are a major public health threat in Asia. The present investigation was carried out to study the ecology of vector mosquitoes in Sri Lanka, in order to explore effective mosquito control strategies in rice ecosystems from the ecological point of view. Method: As part of investigations on potential linkages between irrigation and malaria transmission, all surface water bodies in and around 20 selected villages along an irrigation distributary in the District of Trincomalee, Sri Lanka, were surveyed for anopheline mosquito larvae (Diptera: Culicidae) from January 2012 - June 2013. Breeding sites were characterized according to exposure to sunlight, substratum, and presence of vegetation, fauna, inorganic matter and physical water condition (clear/turbid). Dissolved Oxygen (DO), conductivity, salinity, pH, temperature, Total Dissolved Solids (TDS) and turbidity were recorded. Results: A total of 3914 Anopheles larvae of twelve morphological types were collected from 685 breeding habitats. Anopheles peditaeniatus (n= 1041), was the most abundant, followed by An. subpictus (n= 893), An. nigerrimus (n= 808), An. barbirostris (n= 581), An. pallidus (n= 184), An. annularis (n=124), An. jamesii (n= 80), An. varuna (n= 76), An. vagus (n= 56), An. barbumbrosus (n= 48). An. culicifacies (n= 22) and An. aconitus (n= 1). The four most abundant species were significantly associated with waterlogged paddy fields. Anopheles culicifacies was noted only from irrigational canals with vegetation. Conductivity, salinity, and TDS were positively correlated with larval densities of An. peditaeniatus (Pearson correlations= 0.04, 0.17, 0.33: P= 0.94, 0.77, 0.58), An. subpictus (Pearson correlations= 0.6, 0.68, 0.56: P= 0.19, 0.20, 0.31), and An. vagus (Pearson correlations= 0.6, 0.42, 0.17: P= 0.28, 0.47, 0.77). However all physicochemical variables were negatively correlated with An. culicifacies densities. Conclusion: Major malaria vector of An. culicifacies complex occurred at relatively low densities, mainly in irrigated and waterlogged fields. These findings support understanding of mosquito ecology and will support future mosquito control strategies in rice ecosystems in Sri Lanka.

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