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).
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.
Dengue as-a public health problem in Sri Lanka
(La Fondation pour l’Université de Lyon, 2009) Hapangama, H.A.D.C.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.; Premaratna, R.; Manamperi, A.; Gunasena, S.; Abeyewickreme, W.
Dengue infection is an important global public health problem and an increasing number of persons from the South Asian region have been directly or indirectly affected by the disease. In Sri Lanka, dengue has become a major threat to public health in many urban and sub-urban' areas during past three decades. Rapid unplanned urbanization and increasing human population has increase the rate of infection and the frequency. The study area, Gampaha District is the second most populous district in the country having a population density of 1 539 persons per km2 and was the district reporting the second highest incidence of dengue in 2008. Therefore, current research efforts are focused on dengue transmission, examining the presence of sub-clinical infections, role of vector mosquitoes and Knowledge, Attitude and Practices (KAP) of the community on dengue infection in an effort to contain the disease. In the present study, dengue antibodies were detected in samples collected from clinically suspected patients and as well as in samples collected from volunteers. Volunteer sera collected around the confirmed cases had a 23.6% sero-positive rate for dengue IgM antibodies. The rate of asymptomatic recent infections was calculated to be 16.9%. In present study we have serologically confirmed the presence of subclinical infections and according to the published data this is the first confirmation of asymptomatic dengue infections in Sri Lanka. According to the entomological investigations carried out, the common breeding places for Aedes vectors were found to be discarded small containers. Even though Ae. Aegypti has been considered as the principal vector transmitting dengue fever, current studies highlighted the predominant ro!e of Ae. albopictus in the disease transmission. A previous study in Sri Lanka also suggested that prevalence and .presence of high-density of Ae. albopictus should be considered as a risk factor for endemic/epidemic dengue. In view of the above, the spread of dengue by Ae. albopictus should be a matter of great concern. Findings of KAP survey revealed that the community possessed substantially higher knowledge on the spread of dengue, vectors, vector breeding and also seriousness of the infection. However it was observed that good knowledge does not necessarily lead to good practices. Since the attitudes of the respondents were found to be good and most of them were supportive of control measures; next effort of the present study is to see how a novel community mobilized solid waste management system will be effective in dengue vector control.
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.
Transmission study of dengue fever in Gampaha District, Sri Lanka
(Faculty of Tropical Medicine, Mahidol University, 2007) Hapangama, H.A.D.C.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.; Dayanath, M.Y.D.; Abeyewickreme, W.
RATIONALE: Dengue is an important flaviviral infection in Sri Lanka.OBJECTIVES; To determine the effect of possible contributing factors on transmission of Dengue METHODS: A total of 40 households situated around 7 selected dengue cases confirmed by both IgM-IgG ELISA and HAI assay were taken. A minimum of 3 inhabitants from each household were tested for anti-dengue antibodies. At each household, mosquito surveillance was carried out. RESULTS: Of the 40 households, 26(65%) were positive for dengue viral infection. Of the 148 inhabitants, 41 had evidence of exposure to dengue. Of the 41. asymptomatic dengue infection was observed in 25(61%). Paired sera were collected from 36/41 volunteers and tested by HAI assay which confirmed dengue virus infection in 4(11.1%) and were confirmed secondary. Three (75%) of them were asymptomatic. Balance 32(88.9%) was diagnosed as recent dengue infections. Of that 21(65.6%) were asymptomatic probable secondary 7(33.3%), recent dengue 14(66.6%)]. Clustering of cases was observed in all localities varying from 56.5%. There was a significant statistical association between the presence of a definitive dengue infection with the presence of Aedes vectors (x2=3.1, p=0.1). Ae. albopictus were found in all localities 'while Ae. aegypti was found only in one. Breteau index for Ae. albopictus varied from 0-260 while- container index varied from 0-84%. Highest clustering of dengue cases were observed in localities with the highest indices for' mosquitoes. CONCLUSIONS: Study confirms that the presence of asymptomatic infections and clustering of cases. The predominant vector was Ae. albopictus.
An Entomological study to plan intervention for prevention of dengue in Gampaha district, Sri Lanka
(Elsevier, 2008) Abeyewickreme, W.; Hapangama, H.A.D.C.; Weerasinghe, I.; Hapugoda, M.D.; Gunawardene, Y.I.N.S.; Gunawardena, N.K.; Wickremasinghe, A.R.
BACKGROUND: Dengue is the most common and fastest spreading human arbo viral disease worldwide. Most control efforts are based not on insecticides but on suppression of vectors by reducing the number of larval breeding habitats. METHODS: Total of 2000 houses in the District of Gampaha in the Western Province which had the second highest transmission of dengue in 2006 was selected using grid sampling technique, based on disease incidences during 2003—2006. Mosquito surveillance was carried out from 0900 hrs to 1400 hrs during June-August, 2007. Larval and pupal surveillances were carried out indoors and outdoors using standard larval surveillance techniques. RESULTS: The House, Container and Breteau indices for Aedes larvae were 49.3%, 26.1% and 26.9% respectively. The common breeding places for Aedes species were discarded containers 27.1% (559/2064), unused bowls 16.2% (334/2064), plant axils 11.7% (242/2064), water storing barrels 11.4% (236/2064) and coconut shells 5.4%(111/2064). Most productive container types for larvae were tyres (44.3%), plant axils (37.6%), ceramic jars (36.6%), discarded containers (29.5%) and fish tanks (25.0%). Most productive container types for pupae were discarded containers (13.4%), fish tanks (10.4%), unused bowls (8.38%), ceramic jars (7.31%) and coconut shells (6.3%). Aedes albopictus (larvae-85.3%, pupae-80.4%) was the most predominant vector in the district. Earthen pots, plant axils, discarded containers and buckets were the main sources for Aedes albopictus while tyres, barrels, tanks and bowls were for Aedes aegypti. Further, mixed breeding was observed in water storage containers and tyres. CONCLUSION: Discarded small bowls/containers were the key containers with the highest pupal index. Natural breeding habitats such plant axils may also significantly contribute to pupal production. Therefore an integrated control effort to include community level awareness programmes, improved solid waste management for small containers and efforts to prevent breeding in plants is suggested for prevention of dengue in the district. © 2008 Elsevier Inc.
Silent transmission of the dengue fever in Gampaha District, Sri Lanka
(Malaysian Society of Parasitology and Tropical Medicine, 2007) Hapangama, H.A.D.C.; Gunawardene, Y.I.N.S.; Gunasena, S.; Hapugoda, M.D.; Premaratna, R.; Wellawaththage, L.C.; Abeyewickreme, W.
Dengue fever is a major infectious disease in Sri Lanka. Silent transmission of dengue virus has been suggested as a possible risk factor for the increasing incidence of dengue. The present study was carried out in the District of Gampaha using cluster investigation method. A cluster consisted of a minimum of 20 volunteers (family members and immediate neighbours) of a hospitalized serologically/molecular biologically confirmed dengue patient. Serum samples were collected from 148 volunteers in 7 clusters. Samples were tested for anti-dengue antibodies using Dengue Duo IgM and IgG Rapid Strip Test. Of these, positives were further tested for anti-dengue IgG antibody by Haemagglutination Inhibition (HAI) assay, the gold standard test for serological diagnosis of virus infection. Of the 148, 41 had evidence of exposure to dengue virus by Dengue Duo IgM and IgG Rapid Strip Test [positive for IgM: 28(68.4%), IgM & IgG: 7(17%) and IgG: 6(14.6%)]. Of that 41, paired sera were collected from 36 volunteers and tested by HAI assay which confirmed dengue virus infection in 4(11.1%) [confirmed secondary-4(100%)]. Additional 32(88.9%) were diagnosed as recent dengue infections [probable secondary-17(53.1%), probable dengue- 15(46.9%)]. Out of 36 volunteers, 12(33.3%) were symptomatic [confirmed secondary-1(8.3%), probable secondary-10(83.4%), probable dengue-1(8.3%)] and 24(66.7%) were asymptomatic [confirmed secondary-3(12.5%), probable secondary-7(29.2%), probable dengue-14(58.3%)]. Presence of dengue vectors, Aedes aegypti and/or Aedes albopictus were detected around all 7 clusters. The present study serologically confirms the persistence of silent transmission of dengue virus with a trend towards clustering around cases. Presence of vector species in the area further supports this phenomenon.
Co-existence of double serotypes of dengue in patients of Gampaha District
(Sri Lanka Association for the Advancement of Science, 2007) Jayasooriya, D.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.; Premaratna, R.; Manamperi, A.; de Silva, H.J.; Abeyewickreme, W.
Dengue virus (DENV) known to cause a productive cytolytic infection in humans exists in four different serotypes Dengue 1 (D1), Dengue 2 (D2), Dengue 3 (D3) and Dengue 4 (D4). Among 4 serotypes of DENV, D 3 thought to be associated with explosive DHF epidemics and severe disease in many countries. Our objective was to determine the prevalence of dengue serotypes in Gampaha District and to correlate them with disease severity. Serum samples were collected from patients who were within 4 days of onset of fever and clinically suspected of dengue according to WHO criteria. Total viral RNA extracted from each serum sample was subjected to RT-PCR followed by a semi-nested PCR using specific primers. Out of 91 samples collected between Nov 2005 and Dec 2006, 16 samples were confirmed positive for DENV RNA by RT-PCR. Our results of multiplex semi-nested PCR indicated that 9/16 (56.25 %) of the positive cases were co-infected with serotype 2 and 3 (D2 & D3), while 4/16 (25%) were infected with D 3 and 3/16 (18.75 %) with D 2. 3/4 of D 3 cases had DHF , 1/3 of D2 cases were DHF while there were no DHF cases among the D2 and D3 co-infected patients. The mean Packed cell Volume (PCV) values of D3, D2 and D2 & D3 co-infected were 53.8 %, 48 % and 39.6% respectively while the mean platelet values of those were 66,000 mm3, 123,000 mm3 and 174.000 mm3 , respectively. Dengue infection by a single serotype is common among patients. Although few cases of co-infection by more than one serotype had been previously reported in a few other countries, this is the first description of simultaneous co-infection by D2 and D3 in Gampaha district. In this limited study we have observed a reduction of disease severity in D2 and D3 simultaneously co-infected patients. Could simultaneous co-infection by more than one serotype or a combination of two particular serotypes have lead to a decrease in disease severity among dengue patients is a matter yet to be studied. Further studies are needed to support these conjectures and to establish the clinical implications of simultaneous co-infection on the prevalence of DHF and disease severity. Acknowledgement: NSF (grant SIDA/2006/BT/02) & IAEA (SRL TC 6/028)
A Comparative field study of novel commercial Antigen Detection Enzyme-Linked Immunosorbent Assay (ELISA) with Reverse Transcription Polymerase Chain Reaction (RT- PCR) assay for early definitive laboratory diagnosis of dengue viral infection in Sri Lanka
(Sri Lanka Association for the Advancement of Science, 2007) Hapugoda, M.D.; Jayasooriya, D.H.S.W.; Gunawardene, Y.I.N.S.; Wellawaththage, C.; Premaratna, R.; Abeyewickreme, W.
Dengue is an important mosquito borne viral infection in South East Asia. Early definitive laboratory diagnosis of infection would help in management of patients and reducing the case fatality rate. The objective of this study was to determine the accuracy of novel commercial Antigen Detection Enzyme-Linked Immunosorbent Assay (ELISA) using Non Structural protein 1 (NS1) (Bio Rad) for early definitive laboratory diagnosis of dengue infection under field conditions in Sri Lanka. A panel of acute serum samples collected from 99 patients clinically suspected of having dengue fever (<5 days) warded at the North Colombo Teaching Hospital, Ragama, Sri Lanka were used for the present study. Serum samples were tested using Antigen Detection ELISA according to the method described by the manufacturer. Results of this novel assay were compared with RT-PCR assay using Chi-squared test. Two variables were analyzed at a 95% confidence interval and P value <0.05 was considered as significant. Twenty two and 65 patients were positive and negative, respectively, for dengue infection by both assays. Nine patients were confirmed as dengue by the Antigen Detection ELISA only. Three patients were confirmed as dengue by RT-PCR assay only. Antigen detection ELISA showed 88% of agreement with the RT-PCR assay. According to the Chi-squared test, there was no significant difference between the two assays for early diagnosis of dengue infection (?2=46, P=0.0000). Novel commercial Antigen Detection ELISA kit (Bio-Rad 72830) can be used for early definitive laboratory diagnosis of dengue infection in Sri Lanka under field conditions. Acknowledgement: the International Atomic Energy Agency (SRL 06/28) for technical co-operation and APCOT Marketing LTD, Sri Lanka for supplying Antigen detection ELISA kits.

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