Medicine
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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty
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Item Impact of repeated annual Diethylcarbamazine-Albendazole mass treatment on transmission of Wuchereria bancrofti in the Gampaha district.(Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, 2007) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Abeyewickreme, W.Entomological and parasitological surveys are critical for the baseline evaluation of impact of repeated annual. Diethylcarbamazine-Albendazole mass treatment on the reduction of microfilaria density and level transmission of Wuchereria bancrofti in Culex quinquefaciatus. The objective of this study was to assess the current situation, following the mass drug administration programme in the Gampaha district, with regard to lymphatic filariasis, using entomological and parasitological data. A pilot survey was carried out using parasitological, 'clinical and entomological indicators in 21 sites in 7 Medical Officer of Health areas of Gampaha district to assess the current filariasis situation. The localities were selected from. the Medical Officer of Health areas based on previous data obtained from the regional Anti Filariasis Campaign office. Results indicate that 76.19% (16/21) sites were infested with mosquitoes positive for Wuchereria bancroft and the positivity of 1.44% (31/2157) was observed among the mosquitoes caught from households in the selected sites. The microfilariae wasa determined to be 15.5 per positive mosquito. The parasitological result was indicated 0.017 % prevalence of lymphatic fllariasis in the selected population. Data recorded by the Anti Filariasis Campaign Gampaha in 1994, suggested that significant decrease of infective rate, positivity of mosquito and microfilaria density, which are respectively 90%, 3.05% and 23. Study confirms that active transmission of Wuchereria bancrofti is currently taking place in the Gampaha district, despite, the mass drug administration Programme been implemented since 2002. This study highlights the urgent requirement of a proper screening programme combined with anti filarial treatment and vector control programme to minimize filarial morbidity and interrupt filarial transmission within the country.Item Entomological assesment of Wuchereria bancrofti transmission following mass treatment in Gampaha District, Sri Lanka.(Faculty of Tropical Medicine, Mahidol University, 2007) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Abeyewickreme, W.Global program to eliminate Lymphatic filariasis (Lf) relies mainly on Mass Drug Administration (MDA). Success of filariasis control programs depends on careful monitoring of infection levels in human populations as well as vectors following the introduction of the drug intervention. This study was conducted to assess the current level of Lf transmission following mass distribution of Diethylcarbamazine-Albendazole in the Gampaha district, Sri Lanka. Field study was conducted in 45 sites in all Medical Officer of Health (MOH) areas of the Gampaha district identified by the Anti Filariasis Campaign (AFC) as high risk for bancroftian filariasis. Investigation revealed 42.22% (19/45) of the sites were infested with mosquitoes positive for Wuchereria bancrofti. Only the sites in urban and semi urban areas were positively infested while rural areas were free of infection. An infection rate of 5.26% was observed among the mosquitoes caught from households and the larval density was 8.7 per positive mosquito. According to a study conducted by the AFC in a sample population (14 sites), the prevalence of Lf was 0.038%. Data recorded by the AFC in 1994 indicated that the infective rate, positivity of mosquitoes and microfilaria density were 90%, 3.05% and 23 respectively. The present study confirms that the level of transmission of W. bancrofti has not decreased in the Gampaha district, despite the MDA Programme been implemented since 2002. Therefore, a proper screening-programme combined with anti filarial treatment and. vector control programme is urgently required to minimize filarial morbidity and interrupt filarial transmission within the country.Item Patterns of monthly Culex mosquito density variation in Gampaha district, Sri Lanka.(Faculty of Tropical Medicine, Mahidol University, 2009) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Manamperi, A.; Abeyewickreme, W.BACKGROUND: The ecology, development, behavior, and survival of mosquitoes and the transmission dynamics of the diseases they transmit are strongly influenced by climatic factors. OBJECTIVE: The objective of this study was to identify the population density variation of Culex mosquitoes within a period of one year in Gampaha district, Sri Lanka together with potential climatic factors that influenced the Culex population density variation. METHODOLOGY: Culex mosquitoes were routinely collected on monthly basis from 9 sites in Gampaha district. Climate data was obtained from the Department of Meteorology. RESULTS: An exponential growth of Culex population densities was observed in all sites starting in December to February during the study period. The maximum density occurred in January and decreased from March until July. It again increased during August and thereafter decreased until December. Among the study sites the maximum Culex density (mosquitoes/man-hour) was observed in Hekiththa ranging between 89 to 22 and the minimum was from Kurukulawa ranging from 6 to 1. Climatic data suggest that temperature is a limiting factor for the Culex population growth while it was strongly influenced by the rain fall pattern. DISCUSSION: Similar Culex population density variation pattern was observed in all sites but exhibited enormous variation between sites, probably due to different local conditions. Also it was suggested that estimation of W, bancrofti transmission levels in Culex mosquitoes should be practiced in field settings where high mosquito density was observed. Since mosquito density appears difficult to be analyzed by individual dissection use of pool-screen PCR-ELISA would be a better method.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 Night blood survey of a selected high-risk population for lymphatic filariasis(Sri Lanka Association for the Advancement of Science, 2007) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Abeyewickreme, W.; Gunawardena, N.K.; Hapuarachchi, H.A.C.; Abeysundara, S.Human infection with Wuchereria bancrofti causes a disabling parasitic disease known as lymphatic filariasis, which is a major public health and socio-economic problem in many parts of the world. Little is known about the prevalence of filariasis among high-risk populations for filariasis. Objective of this study was to determine such prevalence of lymphatic filariasis among Mahara prison inmates whom the Anti Filaria Campaign (AFC) has identified as a high-risk group. All inmates of Mahara Prison were screened for Microfilariae (Mf) except those in special cells, by night blood film microscopy to determine the prevalence of infection from February to May 2007. All inmates were males of greater than 15 years. Of the 423 inmates screened, 15 were positive for Mf, giving a Mf positive rate of 3.55% in the study population and a mean Mf density of 5 Mf/60 æl blood, ranging between 4 to 9.2 Mf /60 æl of blood with a standard deviation of 2.49. The highest number of infected inmates was residents of Colombo and Gampaha districts where transmission is currently taking place. This is one of the few studies undertaken to date to determine the prevalence of bancroftian filariasis among inmates of a prison, a neglected population in Sri Lanka. This study indicates that the Mf rate of bancroftian filariasis in this study population is far greater than the 0.18% currently reported in the country. Therefore, an intensive programme is recommended to contain the spread of infection within this study population. For this, a proper screening programme combined with antifilarial treatment and vector control programme is urgently required. Acknowledgements: Authors wish to acknowledge the financial assistance received from WHO/SEARO/TDR (grant no. SN 1152) and University of Kelaniya (Research grant no. RP/03/04/06/01/2006). Authors wish to thank Dr. Ravi Mudaliage, Senior Medical Officer, Prison's Hospital, Mahara, Ragama for his support and encouragement during field study activities. Authors also wish to thank Mr. M. Y. D. Dayanath, Ms. N.M. Ashoka Malanie, Mr. M.I.M.Peris, Mr. Y.L.Rassapana and other staff members of the Molecular Medicine Unit and Department of Parasitology, Faculty of Medicne, University of Kelaniya, Ragama for their assistanceItem Large-scale entomological assessment of Wuchereria bancrofti transmission by dissection and PCR-ELISA in Gampaha district, Sri Lanka(Sri Lanka Association for the Advancement of Science, 2008) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Manamperi, A.; Hapuarachchi, H.A.C.; Bandara, K.B.A.T.; Abeyewickreme, W.Entomological surveys are important tools for monitoring progress of lymphatic filariasis (Lf) eradication programs. In this study, dissection of Culex quinquefasciatus was compared with a Polymerase Chain Reaction - Enzyme Linked Immunosorbent Assay (PCR-ELISA) for pooled mosquitoes to assess filarial infection levels in the major vector of Wuchereria bancrofti in Gampaha district, following mass-treatment programme with diethylcarbamazine (DEC) and albendazole. Mosquitoes were collected in 30 sentinel and 15 non-sentinel sites in 15 Medical Officer of Health (MOH) areas of Gampaha district known for the presence of W. bancrofti transmission. Captured mosquitoes were dissected to determine the W. bancrofti larvae (L1, L2, L3). PCR was carried out using Deoxyribonucleic acid (DNA) extracted from mosquito pools (15 body parts/pool) utilizing primers specific for the 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 (3pools/site) was estimated using the PoolScreenTM algorithm and a novel probability-based method. The prevalence of infected 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% - 25.4%. Mosquitoes collected from all MOH areas (80%, N = 12), except for Minuwangoda, Dompe and Ragama, were positive for W. bancrofti larvae, with a prevalence rate ranging from 0.78% to 16.97% in both methods. Of 30 sentinel sites, 43.3% (N = 13) were positive for W. bancrofti transmission whereas it was evident in 40% (N = 6) of non-sentinel sites. The proportion of positive pools detected by the PCR-ELISA assay was higher than that obtained by the dissection indicating that PCR-ELISA assay is more sensitive than the dissection method in detecting infected/infective mosquitoes. Also results of this study showed that autochthonous transmission of W. bancrofti continues in the Gampaha district despite completion of the 5 year mass drug administration (MDA) programme. Therefore, we emphasize the use of more sensitive tools such as PCR-ELISA to monitor the impact of the MDA programme on disease transmission. This study also emphasizes that control measures should be further continued until the microfilareamic population is reduced to a level which could interrupt transmission in the area. Financial assistance received from WHO/SEARO/TDR (grant no. SN 1152) and University of Kelaniya (Grant no. RP/03/04/06/01/2006) is acknowledgedItem Comparison of five DNA extraction methods from human blood for the detection of Wuchereria bancrofti by polymerase chain reaction assays(Sri Lanka Association for the Advancement of Science, 2008) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Manamperi, A.; Hapuarachchi, H.A.C.; Gunawardena, N.K.; Abeysundara, S.; Abeyewickreme, W.Introduction: Lymphatic filariasis (Lf) is the second most common vector-borne disease globally. Approximately 90% of global burden of Lf is caused by Wuchereria bancrofti. W. bancrofti is routinely diagnosed by morphological identification of microfilariae (Mf) by microscopy which is a labour intense, low sensitive and time consuming method. Detection of W. bancrofti Deoxyribonucleic acid (DNA) using polymerase chain reaction (PCR) technique has become popular today, because of its high sensitivity and specificity. The overall success of the PCR strategy in detecting a filarial parasite in human blood varies between sample preparation methods. The objective of this study was to compare five DNA extraction methods (Lysis + centrifugation, Chelex method, Mf pellet method, Q1Aamp DNA Mini Kit commercial system, and Phenol-chloroform) with regard to duration of completion, labor involvement and PCR analytical sensitivity in-relation to DNA quality and quantity for the detection of W. bancrofti in human blood. Five blood samples positive for mf of W. bancrofti were tested for each DNA extraction method and were compared with respect to the sensitivity, time and quality/quantity of DNA and also by PCR analysis. Of the 5 methods tested, Mf pellet method was found to be the most simple and effective technique for the isolation of W. bancrofti Mf in human blood. This method was quick (15 min to complete), simple (5 min of manual labor), and very economical. It does not require any organic solvents, and the entire extraction procedure uses only two steps requiring supernatant transfer between tubes, hence minimizing the possibility of cross-contamination. Moreover, the PCR analytical sensitivity of the Mf pellet method was comparable to that of the commercial kit used. No PCR inhibitors were detected, independently of Mf count in the blood. Same method (optimal DNA extraction method) can be also used for the detection of parasite DNA from the field collected Mf positive mosquitoes using a PCR. Therefore, we recommend the Mf pellet method for processing large numbers of blood samples in community surveys aimed at determining the prevalence of W. bancrofti infection.Item Prolonged clearence of microfilaraemia in patients with bancroftian filariasis after multiple high doses of ivermectin of diethylacarbamizine(Oxford University Press, 1996) Ismail, M.M.; Weil, G.J.; Jayasinghe, K.S.A.; Premaratne, U.N.; Abeyewickreme, W.; Rajaratnam, H.N.; Sheriff, M.M.R.; Perera, C.S.; Dissanaike, A.S.In a double-blind trial on 37 asymptomatic microfilaraemic subjects (minimum 400 microfilariae [mf] per mL) with Wuchereria bancrofti infection, the safety, tolerability and macrofilaricidal efficacy of 12 fortnightly doses of ivermectin, 400 microg/kg (ivermectin group), was compared with 12 fortnightly doses of diethylcarbamazine (DEC), 10 mg/kg (DEC group), over a period of 129 weeks after treatment. A control group (LDIC group) was treated with low dose ivermectin to clear microfilaraemia, for ethical reasons. Both ivermectin and DEC in high multiple doses were well tolerated and clinically safe. Macrofilaricidal efficacy was assessed by prolonged clearance of microfilaraemia, appearance of local lesions, and reduction of circulating W. bancrofti adult antigen detected by an antigen capture enzyme-linked immunoassay based on the monoclonal antibody AD12. Mf counts fell more rapidly after ivermectin than after DEC, but low residual mf levels were equivalent in these groups after week 4. Conversely, filarial antigen levels fell more rapidly after DEC than after ivermectin, but low residual antigen levels in these groups were statistically equivalent at all times beyond 12 weeks. Mild, self-limited systemic reactions to therapy were observed in all 3 treatment groups. Local reactions, such as development of scrotal nodules, were observed in several subjects in the DEC and ivermectin groups. These results suggested that high dose ivermectin and DEC both had significant macrofilaricidal activity against W. bancrofti, but neither of these intensive therapeutic regimens consistently produced complete cures. Thus, new drugs or dosing schedules are needed to achieve the goal of killing all filarial parasites in the majority of patients.Item Cloning and characterization of a repetitive DNA sequence specific for Wuchereria bancrofti(American Society of Tropical Medicine and Hygiene, 1994) Siridewa, K.; Karunanayake, E.H.; Chandrasekharan, N.V.; Abeyewickreme, W.; Franzen, L.; Aslund, L.; Pettersson, U.A genomic library constructed in a bacteriophage lambda replacement vector (EMBL3) with Wuchereria bancrofti DNA partially digested with Sau 3A I was screened with 32P-labeled W. bancrofti total DNA, and a strongly reactive recombinant, EMBL3Wb34, was isolated. This clone contained an approximately 16-kb insert that showed some cross-hybridization with Brugia malayi and B. pahangi DNA. However, a 969-bp subclone from EMBL3Wb34, designated pWb12, hybridized only with W. bancrofti DNA and was able to detect as little as 300 pg. Furthermore, pWb12 could detect DNA from a single infective larva or one microfilaria. It has a moderate copy number (450-700) and appears to be interspersed within the parasite genome. The nucleotide sequence contains 66% A+T and 34% G+C and shows no notable internal repeats.Item Effect of ivermectin on the development of Wuchereria bancrofti in the vector(Malaysian Society of Parasitology and Tropical Medicine, 1992) Abeyewickreme, W.; Ismail, M.M.; Premaratne, U.N.; Dissanaike, A.S.