Browsing by Author "Wijegunawardana, A.D."
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Item A comprehensive assessment of lymphatic filariasis inSri Lanka six years after cessation of mass drug administration(Public Library of Science, 2014) Rao, R.U.; Nagodavithana, K.C.; Samarasekera, S.D.; Wijegunawardana, A.D.; Premakumara, W.D.; Perera, S.N.; Settinayake, S.; Miller, J.P.; Weil, G.J.BACKGROUND: The Sri Lankan Anti-Filariasis Campaign conducted 5 rounds of mass drug administration (MDA) with diethycarbamazine plus albendazole between 2002 and 2006. We now report results of a comprehensive surveillance program that assessed the lymphatic filariasis (LF) situation in Sri Lanka 6 years after cessation of MDA. METHODOLOGY AND PRINCIPAL FINDINGS: Transmission assessment surveys (TAS) were performed per WHO guidelines in primary school children in 11 evaluation units (EUs) in all 8 formerly endemic districts. All EUs easily satisfied WHO criteria for stopping MDA. Comprehensive surveillance was performed in 19 Public Health Inspector (PHI) areas (subdistrict health administrative units). The surveillance package included cross-sectional community surveys for microfilaremia (Mf) and circulating filarial antigenemia (CFA), school surveys for CFA and anti-filarial antibodies, and collection of Culex mosquitoes with gravid traps for detection of filarial DNA (molecular xenomonitoring, MX). Provisional target rates for interruption of LF transmission were community CFA <2%, antibody in school children <2%, and filarial DNA in mosquitoes <0.25%. Community Mf and CFA prevalence rates ranged from 0-0.9% and 0-3.4%, respectively. Infection rates were significantly higher in males and lower in people who denied prior treatment. Antibody rates in school children exceeded 2% in 10 study sites; the area that had the highest community and school CFA rates also had the highest school antibody rate (6.9%). Filarial DNA rates in mosquitoes exceeded 0.25% in 10 PHI areas. CONCLUSIONS: Comprehensive surveillance is feasible for some national filariasis elimination programs. Low-level persistence of LF was present in all study sites; several sites failed to meet provisional endpoint criteria for LF elimination, and follow-up testing will be needed in these areas. TAS was not sensitive for detecting low-level persistence of filariasis in Sri Lanka. We recommend use of antibody and MX testing as tools to complement TAS for post-MDA surveillance.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 Evaluation of PCR-ELISA as a tool for monitoring transmission of Wuchereria bancrofti in District of Gampaha, Sri Lanka(Asian Pacific Organization for Cancer Prevention, 2013) Wijegunawardana, A.D.; Gunawardene, N.S.; Hapuarachchi, C.; Manamperi, A.; Gunawardena, K.; Abeyewickreme, W.; Latif, B.OBJECTIVE: To compare Wuchereria bancrofti (W. bancrofti) infection rates of Culex quinquefasciatus, using dissection and PCR-ELISA in two consecutive time periods (from 2007 to 2008 and from 2008 to 2009). METHODS: Mosquitoes were collected in 30 sentinel and 15 non-sentinel sites in 15 Medical Officer of Health areas of Gampaha Districtknown for the presence of W. bancrofti transmission in two consecutive time period of 2007 to 2008 and 2008 to 2009. Captured mosquitoes were dissected to determine the W. bancrofti larvae (L1, L2, L3). PCR was carried out using DNA extracted from mosquito pools (15 body parts/pool) utilizing the primers specific for Wb-SspI repeat. PCR products were analyzed by hybridization ELISA using fluorescein-labeled wild type specific probes. The prevalence of infected/infective mosquitoes in PCR pools (3 pools/site) was estimated using the PoolScreen™ algorithm and a novel probability-based method. RESULTS: Of 45 batches of mosquitoes dissected, W. bancrofti infected mosquitoes were found in 19 and 13 batches, with an infection rate of 13.29% and 3.10% with mean larval density of 8.7 and 1.0 larvae per mosquito for two study periods in the Gampaha District. Total of 405 pools of head, thorax and abdomen were processed by PCR-ELISA for each year. Of these, 51 and 31 pools were positive for W. bancrofti in the two study periods respectively. The association of dissection based prevalence rates with PCR based rates as determined by the Pearson correlation coefficient were 0.176 and 0.890 respectively for the two periods. CONCLUSIONS: Data indicate that PCR-ELISA is more sensitive than the traditional dissection techniques for monitoring transmission intensity