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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Item 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).Item 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 controlItem 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.Item 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.Item 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.Item 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.Item Resting behavior of malaria vectors in Mannar District of Sri Lanka with reference to zooprophylaxis(University of Kelaniya, 2013) Gunathilaka, P.A.D.H.N.; Fernando, M.A.S.T.; Hapugoda, M.D.; Wijeyerathne, P.; Wickremasinghe, A.R.; Abeyewickreme, W.Background: The most important factors for effective zooprophylaxis in reducing malaria transmission are the presence of predominate populations of a strongly zoophilic mosquitoes and domestic animals for their blood-meal. Methods: Resting behavior of anopheline mosquitoes was evaluated using indoor and outdoor mosquito collection techniques. Mosquitoes were captured using 389 outdoor cattle-baited traps and indoor hand catches in 5672 houses at 12 possible malaria sensitive areas situated 12 km apart; Pesali, Keeri, Erukkallampeddi, Moor, Vankalai, Narawallikulam, Vanchiyankulam, Arvanum, Arrippu, Kokkupadayan, Koolankulam and Pandaraveilli for 12 months (June 2012 to June 2013) on a weekly basis. These areas had mostly homogenous characteristics in vegetations and house type. All collected mosquitoes were identified using standard taxonomic keys. A hypothesis test was conducted to determine whether there is a difference between indoor and outdoor collections. Results: A total of 3986 anopheline mosquitoes were collected from both indoor (n= 3122) and outdoor (n= 864) collection techniques. Anopheles subpictus was found to account for over 97.5% (n= 3887) of Anopheles species collected in the study areas. Other species were; 1.4% An. peditaeniatus (n= 57), 0.6% An. varuna (n= 22), 0.3% An. pallidus (n= 10), 0.2% An. nigerrimus (n= 7), 0.05% An. tessellatus (n= 2), and 0.03% An. aconitus (n= 1). Only 3 species namely, An. subpictus (n= 485), An. tessellatus (n= 2) and An. aconitus (n= 1) were recorded from indoor hand catches. In experimental cattle-baited hut trials, An. subpictus (n= 3026) was found to have the highest exophilic tendency (96.9%) compared to other anophelines encountered. The hypothesis test suggests that there was a significant difference of An. subpictus attracted to cattle-baited traps and human dwellings (P = 0.008). Although the main vector An. culicifacies was absent, the presence of the secondary vector An. subpictus was observed in high numbers in the District. Conclusion: Exophilic and zoophilic tendencies of An. subpictus are conducive for zooprophylaxis. The study further indicates that zooprophylaxis may be a potential strategy for malaria control. It is recommended that in areas with a predominant An. subpictus population, cattle could be kept close to human dwellings to maximize the effects of zooprophylaxis.Item Breeding of Anopheles culicifacies in different waterbodies in the district of Trincomalee(University of Kelaniya, 2012) Gunathilaka, P.A.D.H.N.; Fernando, M.A.S.T.; Hapugoda, M.D.; Wijeyerathne, P.; Wickremasinghe, A.R.; Abeyewickreme, W.Introduction: Anopheles culicifacies (Diptera: Culicidae), the major vector of malaria in Sri Lanka is known to breed in clean and clear water. This study was focused to understand the larval habitats of the major malaria vector with the eco system changes in the Trincomalee district of the Eastern Province. Method: Potential larval habitats for Anopheles mosquitoes were surveyed on a monthly basis for 17 months (January 2011 –June 2012) in 4 different selected sampling sites (Murthankulam, Kommnaimottai, Paranamadawachchiya and Kokmotawewa). Collected larvae were identified using standard taxonomic keys. The species Distribution (C) and Density (D) were calculated. Results: A total of 2996 larval specimens representing 13 Anopheles species were reported from 16 different breeding habitats namely, waste water (n= 635), built well (n= 1229), earth well (n=149), agricultural well (n=9), rain water collection (n=89), animal hoof print (n=17), burrow pit (n=256), rock pool (n=10), canal (n=15), irrigation canal (n=27), lake margin (n=27), tank margin (n=448), pond margin (n=15), marshy land (n=13), paddy field (n=15) and slow moving water (n=42). An. culicifacies was observed as the most predominant species throughout the survey. According to Density criterion, An. culicifacies (44.0%), An. subpictus (19.2%), An. barbirostris (13.2%), An. peditaeniatus (10.28%) and An. nigerrimus (8.7%) were within the dominant class; (D > 5%). Two species (An. vagus, An. pallidus) were in the subdominant class (1< D <5%). Only An. annularis, An. varuna, An. barbumbrosus, An. pseudojamesi, An. jamesii and An. tessellatus were the satellite species (D < 1%). An. nigerrimus, An. subpictus and An. peditaeniatus can be regarded as constant according to distribution (C= 80.1-100%). Only An. vagus was the most frequently reported (C= 60.1 – 80%) species. All other Anopheles including An. culicifacies were observed as infrequent species (C= 20.1 – 40%) and no species was identified as sporadic appearance (C= 0 – 20%). Most productive breeding site for An. culicifacies were drains covered with waste water (Density= 81.57%) in remote areas. Interpretation & conclusion: These results indicate that An. culicifacies has adapted to breed in a wide range of water bodies including waste water collections although they are considered to breed in clean and clear water. The survival of the major vector mosquito in widespread water bodies could be responsible for the increase in the incidence of malaria in the future.Item Susceptibility of malaria vectors to insecticides in Ampara, Batticaloa, Trincomalee & Mannar districts of Sri Lanka(University of Kelaniya, 2012) Gunathilaka, P.A.D.H.N.; Fernando, M.A.S.T.; Hapugoda, M.D.; Wijeyerathne, P.; Wickremasinghe, A.R.; Abeyewickreme, W.Background: The current status of insecticide resistance was studied for potential malaria vectors in four districts namely Ampara, Batticaloa, Mannar and Trincomalee of Sri Lanka. Methods: Insecticide-susceptibility tests were carried out using WHO standard kits against various chemical compounds at ambient room temperature of 27+1oC and relative humidity of 75-80% for adult and larvae separately. A total of 3629 adult female Anopheles mosquitoes belong to nine species (An. subpictus, An. vagus, An. nigerrimus, An. peditaeniatus, An. pallidus, An. annularis, An. jamesii, An. pseudojamesi and An. barbirostris) were exposed to cyfluthrin- 0.15%, etofenofrox- 0.5%, bendiocarb- 0.01%, malathion – 5%, deltamethrin – 0.05%, permethrin – 0.75%, λ- cyhalothrin– 0.05%, fenitrothion – 1.0% and propoxur – 0.1%. A total of 610 larvae belong to five Anopheles species (An. subpictus, An. vagus, An. barbirostris and An. peditaeniatus) were exposed to 0.0025, 0.005, 0.125 and 0.625 mg/l of viz. temephos (Abate). Results: All of the mosquito larvae or adult species used for the study were susceptible for selected insecticides (Mortality 98-100%). Adult Anopheles species showed a possibility of developing resistance to some chemicals (Mortality 97- 80%): An. nigerrimus (lambdacyhalothrin and permethrin), An. peditaeniatus (malathion, deltamethrin and cyfluthrin) An. subpictus (deltamethrin, permethrin, propoxur, cyfluthrin, and etofenprox), An. vagus (deltamethrin, permethrin and etofenprox) and An. pallidus (deltamethrin). Anopheles larvae of An. subpictus, An. barbirostris and An. peditaeniatus showed the potential of developing resistance (Mortality 97-80%). Interpretation & conclusion: As a result of resettlements and expanding commercial agriculture there can be a potentiality of developing insecticide resistance in mosquito individuals. Therefore, proper use of chemicals as pesticides should be adopted in these areas.Item Blood-feeding patterns of Anopheles mosquitoes in malaria-endemic areas of Sri Lanka(University of Kelaniya, 2012) Gunathilaka, P.A.D.H.N.; Fernando, M.A.S.T.; Hapugoda, M.D.; Wijeyerathne, P.; Wickremasinghe, A.R.; Abeyewickreme, W.Background: Studies on host preference patterns in blood-feeding of anopheline mosquitoes are crucial for incriminating them as malaria vectors. However, little information is available on the host preferences of Anopheles mosquitoes in Sri Lanka. Therefore, the objective of the present study was to determine the hematophagic tendency of the anophelines. Methods: Adult Anopheles mosquitoes were collected using Cattle Baited Trap Collection (CBTC), Cattle Baited Net Collection (CBNC), Window Trap Collection (WTC), and Hand Collection (HC) from selected sentinel sites in Mannnar (3) and Trincomalee (5) Districts during June 2011- June 2012. Each blood fed mosquito was processed in to 9 cm whatman filter papers within 24 hours after blood meal has taken. DNA was extracted using the dried blood meal protocol of the QIAmp DNA mini kit. A multiplexed, Real Time Polymerase Reaction (RT- PCR) assay targeting 8 animals was developed for two panels (Panel 1: Bovine, cat, pig, monkey: Panel 2: Human, rat, dog, chicken) to identify the host meal of Anopheles. Human Blood Index (HBI), Forage Ratio (FR) and Host Feeding Index (HFI) were calculated. Results: A total of 216 field caught freshly engorged females mosquitoes belonging to 12 Anopheles species was analyzed. The host preference of anophelines observed in this study was bovine (86.17%), human (1.84%), cat (0.46%) and pig (0.46%). Only 6.91 % was positive for both human and bovine. In addition 5.0 % of the total samples tested were unknown. The overall HBI and HFI in the present study were low indicating the humans were not the preferred host for the tested anopheline species. Nevertheless, a small proportion engorged An. aconitus (0.37), An. culicifacies (0.27), An. barbirostris (0.2), An. annularis (0.125) and An. subpictus (0.12), An. peditaeniatus (0.08), An. pseudojamesi (0.04) and An. barbumbrosus (0.04) contained human blood, The FRs for human were <1.0 for most of the anophelines, except An. aconitus (1.04). Conclusion: The presence of human blood, in mosquito species indicates the possibility of them transmitting malaria. Hence, further studies on vector competence are needed to determine the role of each of the above anopheline species currently as efficient vectors of malaria.