Browsing by Author "Abeyewickreme, W."
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Item Aedes albopictus the “underrated” Asian Tiger(University of Kelaniya, 2010) Jayasooriya, D.H.S.W.; Gunawardene, Y.I.N.S.; Manamperi, A.; de Silva, H.J.; Abeyewickreme, W.Introduction The mosquito Aedes aegypti was thought to be the main vector responsible for virtually all dengue epidemics; while Aedes albopictus was considered a vector in which the virus is maintained but does not cause epidemics. Objective The study was conducted covering three endemic districts in Sri Lanka to determine the role of genus Aedes during dengue transmission. Methods and Material Mosquitoes were collected within a 350m radius from the location of the positive patients. Heads and abdomens of 63 pools were tested for DENV RNA with and RT-PCR-LH-(P32) assays Results Discussion Ae. albopictus was present in majority of the locations in all districts surveyed. Ae. albopictus was found in 13/17 (76.47%), 24/25 (96%)and 19/22 (86.36%) sites in Colombo, Gampaha and Kurunegala respectively. The RT-PCR-LH-(P32) assays indicated that 5/25 (20%) sites in Gampaha, 2/17 (11.76%) in Colombo and 6/22 (27.27%) in Kurunegala were positive for DENV. In Gampaha and Colombo there were 3 and 1 of DEN-2 positive pools respectively, while there were 2 and 1 of DEN-3 positive pools respectively. A higher number of positive pools (4/1or 21.05%) for DEN-1 and 1/1(5.26 %) for DEN-4 were found in Kurunegala. In Kurunegala one pool was positive for both DEN-2 and DEN-4 indicating the circulation of multiple serotypes within close proximity. Moreover one of the three DEN-2 positive pools in Gampaha consisting of only male Ae. albopictus mosquitoes is supportive of the belief of vertical transmission of DENV. In a DEN-4 positive location in Kurunegala HI was found to be10%, BI= 1and CI= 5.88 %while anotherDEN-2 positive site in Wattala showed HI of 5.55%and a BI of 5.55 suggesting active transmission. The abundance of Ae. albopictus in all districts and the findings indicating that100% of the positive pools were made of Ae. albopictus in this study highlights the importance of Ae. albopictus in the transmission dynamics dengue. The ability of Ae. albopictus to be infected with low viremia and the degree to which it permits replication within the mosquito itself could have an impact on the transmission and these verity of the disease. Co-circulation of two or more serotypes in a single pool or in different pools of mosquitoes within the same district is suggestive of hyper endemic transmission dengue in the three districts. The greater susceptibility of Ae. albopictus to infection by DENV is said to lead to greater virus adaptation. Sri Lanka as a whole would be at serious risks for multiple outbreaks in future. Our results indicate that Ae. albopictus is more efficient in dengue transmission than previously thought. The results shed light on the efficiency of Ae. albopictus as a vector in transmitting DENV in the absence or low abundance of Ae. aegypti in Sri Lanka. The present study suggests that Ae. albopictus sp is underrated in terms of transmission potential during peak transmission periods of dengue in Sri Lanka. Key words: RT-PCR-LH-(P32) RT-PCR-Liquid Hybridization with P32 radio isotope, HI-House hold Index, BI- Breteau Index, CI-Container Index,DENV-Dengue Virus Authors wish to acknowledge the financial assistance rendered by the NSF Sri Lanka (GrantNo:SIDA/2006/BT/02)and the IAEA (Grant NoTC SRL 6/028).Item Alteration in Aedes togoi susceptibility to Brugia pahangi microfilariae induced by Aedes albopictus thoracic homogenate(SEAMEO Regional Tropical Medicine and Public Health Project, 1989) Abeyewickreme, W.; Sucharit, S.; Choochote, W.; Chaicumpa, W.; Tumrasavin, W.Refractoriness to Brugia pahangi microfilarial infection could be induced in the normally susceptible Aedes togoi mosquitoes by intrathoracic injection with crude thoracic homogenate of the refractory Aedes albopictus mosquitoes. The crude thoracic homogenate contained three Sephadex G-200 protein profiles of which the first profile showed strongest inhibition to the parasite development.Item Anopheles (Cellia) culicifacies: a potential vector of bancroftian filariasis in Sri Lanka(Museum and Reference Centre, SEAMEO-TROPMED National Centre of Thailand, 1987) Abeyewickreme, W.; Hewadikaram, K.A.; Weerasena, K.H.; Premaratne, U.N.; Ismail, M.M.Item Anopheles (Cellia) jamesii: a potential natural vector of Bancroftian filariasis in Sri Lanka(Oxford University Press, 1991) Abeyewickreme, W.; Wanniarachchi, P.No Abstract AvailableItem Anopheles (Cellia) tessellatus; an efficient laboratory vector of Bancroftian filariasis in Sri Lanka(Museum and Reference Centre, SEAMEO-TROPMED National Centre of Thailand, 1985) Abeyewickreme, W.; Ismail, M.M.Item Anopheles culicifacies breeding in polluted water bodies in Trincomalee district of Sri Lanka(BioMed Central, 2013) Gunathilaka, N.; Fernando, T.; Hapugoda, M.; Wickremasinghe, R.; Wijeyerathne, P.; Abeyewickreme, W.BACKGROUND: Anopheles culicifacies, the major vector of malaria in Sri Lanka, is known to breed in clean and clear water. The main objective of the study was to detect the breeding habitat diversity of An. culicifacies. METHODS:Potential larval habitats for Anopheles mosquitoes were surveyed on a monthly basis for 17 months (January 2011--June 2012) in four different selected sampling sites (Murthankulam, Kommnaimottai, Paranamadawachchiya and Kokmotawewa) in Trincomalee District of Sri Lanka. RESULTS: A total of 2,996 larval specimens representing 13 Anopheles species were reported from 16 different breeding habitats. According to density criterion, An. culicifacies, Anopheles subpictus, Anopheles barbirostris, Anopheles peditaeniatus and Anopheles nigerrimus were dominant. Anopheles nigerrimus, An. subpictus and An. peditaeniatus were observed as constant in relation to their distribution. The most productive breeding site for An. culicifacies was drains filled with waste water in remote areas; the second highest productivity was found in built wells. CONCLUSIONS: These results indicate that An. culicifacies has adapted to breed in a wide range of water bodies including waste water collections although they were earlier considered to breed only in clean and clear water. Copyright © 2013 Gunathilaka et al.; licensee BioMed Central Ltd.Item Anopheles subpictus s.l. breeding in polluted water bodies in Vankalai area in the Mannar District(Sri Lanka Association for the Advancement of Science, 2014) Ranathunge, R.M.T.B.; Gunathilaka, P.A.D.H.N.; Kannangara, D.N.; Abeyewickreme, W.; Hapugoda, M.D.Item Appearance of Anopheles jeyporiensis James from Sri Lanka; a new species to the mosquito checklist(The Japan Society of Medical Entomology and Zoology, 2015) Gunathilaka, N.; Abeyewickreme, W.; Hapugoda, M.; Wickremasinghe, A.R.Previous records of Sri Lankan Anopheles were most imperfect, and even the number and names of the species present were very doubtful. There was no systematic study conducted for Sri Lankan adult anophelines since 1990. Therefore, the objective of this study was to explore the species abundance and morphological variations of anopheline mosquitoes in Sri Lanka. Entomological surveys were conducted on a monthly basis from June 2010 to December 2013 in Trincomalee District, using five entomological techniques. Entomological surveys identified a total of 131,804 mosquito specimens belong to 18 anopheline species. One of which was An. jeyporiensis, a species that was not in the checklist in Sri Lanka. Its basic morphological features are similar to the members in Myzomyia series under the subgenus Cellia. Following characteristics were used to confirm the species as An. jeyporiensis; Centre of the scutum covered with short oblong white scales extending back to scutellum; Vein R 1 usually with accessory pale spot on preapical dark (PD) area; Foretarsomere 1 with apical pale band nearly 2.0 width of tarsomere diameter.Item Application of a real time Polymerase Chain Reaction (PCR) assay for the early diagnosis of human leptospirosis in Sri Lanka(Academic Press, Elsevier, 2016) Denipitiya, D.T.H.; Chandrasekharan, N.V.; Abeyewickreme, W.; Hartskeerl, C.M.; Hartskeerl, R.A.; Jiffrey, A.M.; Hapugoda, M.D.Leptospirosis has a major impact on health in Sri Lanka but is probably grossly under-recognized due to difficulties in clinical diagnosis and lack of diagnostic laboratory services. The objective of this study was to establish and evaluate a SYBR Green-based real-time Polymerase Chain Reaction (rt-PCR) assay for early, rapid and definitive laboratory diagnosis of leptospirosis in Sri Lanka. The rt-PCR assay was established and analytical specificity and sensitivity were determined using reference DNA samples. Evaluation of the assay for diagnosis of clinical samples was performed using two panels of serum samples obtained from 111 clinically suspected adult patients. Patients were confirmed as leptospirosis (n = 65) and non-leptospirosis (n = 30) by the Patoc - MAT. Other 16 samples gave ambiguous results. The analytical sensitivity of the rt-PCR was approximately 60 genome copies and no cross-reactivity was observed with saprophytic Leptospira spp. and other pathogenic microorganisms. Based on confirmation with Patoc-MAT on paired samples this corresponds to a diagnostic sensitivity and specificity of 67.7% (44/65) and 90.0% (27/30), respectively. This study showed that rt-PCR has the potential to facilitate rapid and definitive diagnosis of leptospirosis during early phase of infection in Sri Lanka.Item Application of a Real Time Polymerase Chain Reaction (PCR) for Detection of Pathogenic Leptospira in Clinical Samples(University of Kelaniya, 2012) Denipitiya, D.T.H.; Jiffrey, A.M.; Abeyewickreme, W.; Wellawaththge, C.; Hapugoda, M.D.Leptospirosis, is a zoonotic disease with worldwide distribution, caused by pathogenic species of the genus Leptospira. It has the greatest impact on health in developing countries where it is often grossly under-recognized. Clinical features are similar to a range of other infectious diseases that occur in the same environmental and climatologic conditions. Therefore, laboratory confirmation is essential for proper management of leptospirosis patients. Molecular assays offer definitive laboratory confirmation of leptospirosis at the early phase of infection (1-5 days of fever) within a few hours. The objective of this study was to establish and evaluate potential use of a real time- PCR assay for early, definitive laboratory confirmation of leptospirosis patients. A SYBR green-based real time PCR assay targeting a 203 bp fragment on the secY gene which is conserved among pathogenic serovars of Leptospira was established using a reference DNA sample (Leptospira interrogans strain RGA). Analytical specificity of the assay was tested with the DNA from pathogenic and non-pathogenic Leptospira spp. and five other micro organisms. Analytical sensitivity of the assay was tested using serial dilutions of the reference sample. A panel of acute blood samples (n=150) collected during early phase of infection (1-5 days of fever) from leptospirosis suspected patients was used for evaluation of real time PCR vs qualitative PCR. The results show, real time PCR assay with high analytical specificity (100%) was established and the assay shows 100 times higher sensitivity over qualitative PCR assay (1.3 pg/ml). Real time PCR and qualitative PCR could diagnose current leptospirosis infection in 37.3% (56/150) and 19.3% (29/150) suspected patients respectively. These results indicate high sensitivity of real time PCR over qualitative PCR for diagnosis of leptospirosis patients. In conclusion, this study shows that real time PCR has the potential to facilitate rapid and sensitive diagnosis of acute leptospirosis during early phase of infection.Item Application of nucleic acid technology (NAT) in the diagnosis of active viral replication in HBV and HCV infections and evidence for HBV surface antigen mutants(Sri Lanka Association for the Advancement of Science, 2008) Manamperi, A.; Gunawardene, Y.I.N.S.; Hapuarachchi, C.; Bandara, A.; Wellawaththage, C.; Abeyewickreme, W.; de Silva, J.Introduction: The community prevalence of Hepatitis B (HBV) and hepatitis C (HCV) infections, although considered low (< 1%) in Sri Lanka based on serological markers, pose a significant health threat to patients in high risk groups. The early diagnosis of active viral infection is crucial in such situations to prevent further transmission and to enable the clinicians to initiate successful therapeutic interventions. Objective: This study was carried out to investigate the usefulness of polymerase chain reaction (PCR) in the diagnosis of active viral replication in HBV and HCV infections. Methodology: All specimens from patients with serological evidence of hepatitis B (HBV surface antigen and/or antibodies for HBV core protein) or hepatitis C (antibodies for hepatitis C core protein-Anti-HCV) and referred to the Molecular Medicine Unit from May 2005 to May 2008 were analyzed by PCR and reverse-transcription PCR (RT-PCR) for HBV DNA (n=130) and HCV RNA (n=95) respectively. Results: Of the 130 patients tested, 57 (44%) were positive for HBV DNA. The positive group of patients included 10 renal transplant patients, 4 multiply transfused patients, 4 paediatric patients with lymphoma, and 1 patient with cirrhosis. Six HBV DNA positive patients had negative HBsAg serology profiles indicating the possibility of surface antigen mutant strains. The HBV DNA negative patients with positive serology profiles indicate sero-converted/ patients with resolved infections or false positive serology results. Of the 95 patients tested, 14 (15%) were positive for HCV RNA and included 3 paediatric patients with thalassaemia. HCV RNA negative, anti-HCV positive profiles reflect either false positive serology results (due to less specific antibody assays) or donors who have been exposed to HCV previously and subsequently resolved their infections. Conclusions: A major proportion of patients with serological markers for HBV have active viral infection whereas only relatively a minor proportion of patients with serological markers for HCV have active viral replication. We have also found the first possible evidence of hepatitis B surface antigen mutant strains. This underlines the importance of the nucleic acid based technology in the diagnosis and assessment of infection with or suspected to have hepatitis B or C infections. We also emphasize the importance of introducing NAT for screening donors for HBV DNA and HCV RNA to substantially lower the risk of acquiring HBV/HCV infection from a transfusion.Item Assessment of artificial blood feeding methods and efficacy of different blood meal sources in mass rearing of Aedes aegypti for innovative vector controlling strategies(Faculty of Graduate Studies, University of Kelaniya, Sri Lanka, 2016) Gunathilaka, P.A.D.H.N.; Ranathunga, R.M.T.B.; Udayanga, N.W.B.A.L.; Abeyewickreme, W.Background Mass rearing of vector mosquitoes of dengue is a crucial requirement for implementation of innovative vector controlling strategies such as Sterile Insect Technique (SIT), Incompatible Insect Technique (IIT) and transgenic mosquitoes. Selection of the artificial membrane-feeding technique and blood meal source have been recognized as key considerations in mass rearing of vectors. Methodology Three artificial membrane feeding techniques (Glass plate method, Metal plate and Hemotek membrane feeding method) and three blood sources (human, cattle and chicken) were evaluated based on feeding rates, fecundity and hatching rates of Ae. aegypti, under laboratory conditions. One-way ANOVA, cluster analysis, analysis of variance (ANOSIM) and principal coordinates (PCO) analysis, were used to investigate the significance in the variations among blood feeding. Results Feeding rates of Ae. aegypti significantly differed among the membrane feeding techniques as suggested by one-way ANOVA (p <0.05). The metal plate method was identified as the most efficient and cost effective feeding technique. Blood feeding rate of Ae. aegypti was higher with human blood followed by cattle and chicken blood, respectively. However, no significant difference was observed among mosquitoes fed with cattle blood and human blood, in terms of fecundity, oviposition rate and fertility. Conclusions Metal plate feeding technique could be recommended as the most effective membrane feeding technique for mass rearing of Ae. aegypti, due to its high feeding rate and cost effectiveness. Since, obtaining human blood for insectary colony maintenance is problematic due certain ethical issues, cattle blood could be recommended for mass rearing Ae. aegypti.Item Assessment of possible social and entomological risk factors affecting transmission of dengue in the District of Gampaha(Faculty of Graduate Studies, University of Kelaniya, 2015) Withanage, G.P.W.K.; Abeyewickreme, W.; Hapugoda, M.D.Dengue is the most important mosquito-borne viral infection transmitted to humans in Sri Lanka causing more than 30,000 cases annually. The objective of the study was to identify possible social and entomological risk factors affecting transmission of dengue in Gampaha district where the second highest number of dengue cases recorded during last ten years. Four Medical Officer of Health (MOH) areas were selected based on annual number of dengue cases greater than 250 for last ten years. One GramaNiladhari (GN) division was selected from each MOH area as a study area where the highest number of dengue incidents reported. In each study area, a cluster of 150 households was selected and household and entomological surveys were conducted. The selected areas were Eriyawatiya (Kelaniya MOH), Welikadamulla (Wattala MOH), Akbar town (Mahara MOH), and 3-Kurana (Negombo MOH) GN divisions. There were 2577 population in 600 households in the study areas and 53.5% were females. Average size of a homestead was 17 perches and most of the households (98%-588/600) were individual house. Main vegetations in the homesteads were grass and bushes (97%-583/600) and potential breeding places of dengue vector mosquitoes were observed in 96.1% (577/600) households. Main dengue vector mosquito present in the captured Aedes adult mosquitoes was Aedes albopictusis (92.9%-183/197). Most prominent breeding places were discarded bottles and tins (4.0%-15/371), plant axils (9.7%-36/371), plastic containers (26.4%-98/371), and roof gutters (4.3%-16/371) in the observed premises and 9.2% (34/371) were Aedes larvae positive. Most frequent mosquito preventive measure was bed-nets (30.3%-182/600), but mosquito coils (30.8%-185/600) and vaporizers (17.6%- 106/600) were frequently used. Participants have adequate knowledge about the disease, but they neglect preventive measures and highly depend on government vector control programs. Unplanned households, crowded conditions, poor waste management systems, and negligence to pursue preventive measures may be possible risk factors and therefore, frequent encouragement is needed to control dengue transmission.Item Awareness on dengue and solid waste management among selected school students in Gampaha District of Sri Lanka(Moleclar Medicine Unit, Faculty of Medicine, University of Kelaniya, Sri Lanka, 2015) Radhika, N.M.L.; Gunathilaka, P.A.D.H.N.; Abeyewickreme, W.; Kasturiratne, K.T.A.A.BACKGROUND: Community mobilization is often respected as a powerful tool in ensuring the effective controlling of dengue via promoting effective and sustainable dengue vector control strategies. The current study attempts to investigate the level of awareness on dengue and solid waste management, among selected school students in Gampaha District which is the second highest dengue reported district in Sri Lanka. METHODOLOGY: Awareness programmes on dengue (transmission, vector ecology and preventive measures) and solid waste management were conducted in 10 schools belong to Kelaniya Medical Officer of Health (MOH) area selecting a minimum group of 200 students from each school within the age of 13 - 15 years. Tools such audio-visual presentations, leaflets and videos were used for the awareness programmes. A questionnaire was given to each student prior to the awareness and after the awareness programs as pre and post evaluations respectively in order to assess the general awareness on dengue and solid waste management before and after the awareness programme. One-Way ANOVA, paired t-test and multiple regression techniques were utilized in evaluation of the degree of awareness and key factors affecting the level of awareness, prior and after the conducting the awareness programmes. RESULTS: The degree of initial awareness on dengue and solid waste management of the target school community was recognized to be significantly characterized by the grade of the students and the Education Zonal Category [EZU] (into which their school belongs to) in accordance with One-Way ANOVA (p < 0.05). A significant increment in post awareness of the target school community was recognized due to the conducting of awareness programmes in accordance with the statistics of paired t-test (p < 0.05). Furthermore, based on the multiple linear regression modelling, the post awareness was significantly influenced by the grade of the students and the initial awareness (p < 0.05), regardless of the EZU (p > 0.05). CONCLUSIONS: The awareness of school students on dengue and solid waste management is significantly affected by the grade of the students and EZCs. Awareness programmes are an effective media of raising the awareness of school communities on dengue and solid waste management, while motivating them to actively contribute for the controlling of dengue, via establishing a strong inter sectorial network among schools and other relevant institutions within the Gampaha District.Item Biocontrol potential of six locally available fish species as predators of Aedes aegypti in Sri Lanka(Elsevier Ltd, 2021) Ranathunge, T.; Kusumawathie, P.H.D.; Abeyewickreme, W.; Udayanga, L.; Fernando, T.; Hapugoda, M.ABSTRACT: This study was conducted to evaluate the potential of six locally abundant fish species to control Aedes mosquito larvae and thereby manage dengue epidemics in a sustainable, cost-effective and environmentally friendly manner. The biocontrol efficacy of six larvivorous fish species, namely, Poecilia reticulata, Rasbora daniconius, Aplocheilus dayi, Oriochromis mossambicus, O. niloticus and Puntius bimaculatus, was evaluated under laboratory and field conditions. Five size-matched fish (of the same species) were introduced into separate tanks (replicates) containing 2 L of dechlorinated water and 200 third instar larvae of Aedes aegypti (L.). The number of larvae consumed by each fish species was recorded at three-hour intervals for 24 h. Acclimatized fish were introduced into a total of eighteen artificial breeding habitats located in the Gampola Medical Officer of Health (MOH) area at the species level with three replications. In addition, three breeding sites without fish were monitored as controls. Aedes larvae were monitored by dipping and siphoning methods in each breeding habitat at weekly intervals for three months and the number of fish surviving in each habitat was tallied. Over 24 h under laboratory conditions, O. mossambicus showed the highest predation rate, consuming 320.2 ± 14.5 larvae per day, with a predatory efficiency of 87.5 ± 3.5%. In comparison, O. niloticus consumed 264.6 ± 12.2 larvae per day with consumption efficiciency of 78.1 ± 3.7%, whereas R. daniconius had the lowest larval consumption (33.2 ± 2.7 larvae per day) and predatory efficiency (33.2 ± 3.2%). Over 12 weeks of observation under field conditions, breeding sites with Ap. dayi had the lowest Aedes larval counts, followed by Po. reticulata. Considering predation efficiency and survival under field conditions, Ap. dayi and Po. reticulata were considered to be the best potential candidates for biological control of Ae. aegypti. Further studies under field settings are warranted to evaluate the survival and predatory potential of the selected candidates under more varied environmental conditions.Item Biology, bionomics and life-table studies of Anopheles stephensi (Diptera: Culicidae) in Sri Lanka and estimating the vectorial potential using mathematical approximations(Elsevier, 2023) Jude, J.; Gunathilaka, N.; Udayanaga, L.; Fernando, D.; Premarathne, P.; Wickremasinghe, R.; Abeyewickreme, W.Background: Anopheles stephensi is an invasive mosquito in Sri Lanka that can potentially transmit malaria. The transmission intensity is linked with biology, bionomic and behavioral aspects of a vector that are associated with the Vectorial Capacity (VC). However, the influence of larval conditions eventually affects the vectorial potential of An. stephensi are not well understood. Methods: A colony of An. stephensi was established at the Regional Centre of the Open University of Sri Lanka, Jaffna District. The colony was maintained under confined conditions according to standard protocols. Biotypes of An. stephensi were characterized by referring to the number of egg ridges. Information on (a) biological aspects of eggs (duration for egg hatching, egg development and hatchability), (b) larval development time, larval survivorship pupation success, resting depth of larvae), (c) pupae (adult emergence rate, average time for adult emergence) and (d) adults (biting frequency, mating success gonotrophic cycle, fecundity, duration for egg-laying, percentage of sexes, adult survival/longevity) were evaluated under life-table analysis. Further, selected morphometric characters of each life cycle stage were recorded from the eggs (length and breadth), larvae (head length, width of head, length of thorax, width of thorax, length of abdomen, width of abdomen, and the total length of larvae), pupae (cephalothoracic length and width) and adults (length & width of wing, thorax and abdomen). The VC was calculated using a mathematical-based approach. Descriptive statistics, General Linear Model (GLM) and independent-sample t-test were used for the statistical analysis. Results: All three biotypes were identified based on egg morphology. Mysorensis biotype (47%; n = 470) was predominant followed by type (38.1%; n = 381) and intermediate (14.9%; n = 149). The mean egg length (F(2,997) = 3.56; P = 0.029) and breadth (F(2,997) = 4.57; P = 0.011) denoted significant differences among the three biotypes. The mating success of females observed was 80.7 ± 4.45%. The mean hatching period was 1.9 ± 0.03 days, with a hatching rate of 86.2 ± 0.77%. Overall, 8.0 ± 0.14 days were required for larval development and 30.3 ± 0.14 h were spent in the pupal stage. The pupation success was 94.5 ± 0.37%, and the majority were males (53.1 ± 0.73%). The mean fecundity was 106.5 ± 6.38 eggs and a gonotrophic cycle of 3.4 ± 0.06 days. The female survival rate was 43.2 ± 2.4%, with a mean biting frequency of 66.6 ± 3.5%. The average VC of adult An. stephensi was estimated to be 18.7. Conclusions: The type biotype, which is an effective vector in the Indian subcontinent is present in Sri Lanka. According to the mathematical approximation, An. stephensi found locally has a vectorial capacity of over 18. Therefore, this study warrants the health authorities and vector control programmes to continue the entomological surveys, monitoring of vector densities and implementing appropriate vector control interventions based on biology and bionomic information of vectors.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.Item Breeding habitat diversity and species composition of Anopheles mosquitoes in Trincomalee district, Sri Lanaka(HABITATS Conservation Initiative, 2014) Gunathilaka, P.A.D.H.N.; Fernando, M.A.S.T.; Hapugoda, M.D.; Wijeyerathne, P.; Wickremasinghe, A.R.; Abeyewickreme, W.Entomological studies on the abundance of malaria vector Anopheles mosquitoes have not been studied in some malaria endemic areas of Sri Lanka over past 30 years in view of the security situation. The aim of this study was to explore the habitat diversity and distribution of anopheline species in Trincomalee District in order to prioritize vector breeding habitats for developing timely and cost effective larval controlling measures. Potential larval habitats for Anopheles mosquitoes were surveyed from June 2010 - December 2010, in selected sampling sites in the Trincomalee District; Gomarankadawala, Echchallampaththu, Mollipothana, Thoppur and Padavisiripura, within a radius about 20 km on weekly basis. The species distribution and density were calculated. A total of 3,701 larval specimens representing twelve Anopheles species were reported form 19 breeding habitats (Tank margin, main canal, paddy field, vegetative canal, lake, built well, burrow pit, distribution canal, pond, rock pool, canal, un-built well, common well, river margin, sand pool, animal foot print, rain water collection, quarry pit and marshy land). Ten habitats were categorized under structurally complex group based on the presence of biotic communities. Only An. subpictus can be regarded as constant according to Distribution (C) (C= 80.1-100%).An. nigerrimus, An. peditaeniatus, An. pallidus and An. vagus were frequent (C= 60.1 – 80%). An. varuna, An. barbirostris, An. annularis and An. barbumbrosus were shown as infrequent species (C= 20.1 – 40%) and other namely An. aconitus, An. culicifacies and An. jamesii can be categorized under sporadic appearance (C= 0 – 20%). According to Density (D) criterion, five species (An. subpictus, An. nigerrimus, An. varuna, An. pallidus, An. barbumbrosus) were within the dominant class (D > 5%). Four species (An. vagus, An. peditaeniatus, An. annularis, An. aconitus) were in the subdominant class (1< D <5%). Only An. jamesii and An. culicifacies were the satellite species (D < 1%).Item Breeding of aedes Aegypti and Aedes albopictus in some dengue endemic areas.(Sri Lanka College of Microbiologists, 2000) Hapugoda, G.P.G.M.D.; de Silva, N.R.; Abeyewickreme, W.Dengue fever (DF)/Dengue haemorrnagic fever (DHF) is now- the most important and rapidly spreading vector borne disease in the world. Since 1956, over 350 000 patients have been hospitalized and nearly 12 000 deaths have been reported. In Sri Lanka the incidence of DF/DHF has increased cyclically since the first outbreak in Sri Lanka during which 26 deaths were reported. Aedes aegypti is classified as the predominant vector of dengue in Sri Lanka. Ae, albopictus is considered as an important vector in the absence of Ae. aegypti. In this study, larval surveillance was carried out in fixed monitoring stations / hot-spots and random monitoring stations. Fixed monitoring stations were selected based on high incidence of DF/DHF recorded since 1996 in Kurunegala district. Ten premises within one fixed monitoring station were checked for mosquito breeding weekly using ovitraps and the average monthly ovitrap index (%) was calculated. During outbreaks larval surveillance was conducted in fifteen random monitoring stations including 66 houses which were selected based on serologically confirmed DPI DHF cases in and around Kurunegala and Ragama. Observations on average monthly ovitrap index (%) in the fixed monitoring stations showed that the highest ovitrap index was in Kurunegala town area, Ovitrap index of Ae. albopictus was higher than of Ae. aegypti all localities in and around Kurunegala throughout the study period. Data obtained from random monitoring-stations in and around Kurunegala and Ragama revealed that only Ae. albopictus larvae were present in seven stations. There were no stations in which only Ae.aegypti larvae were present. House index of Ae. albopictus was 28% whereas it was 10.6% for both species in random monitoring stations. Results suggest that Ae.albopictus may play a major role in transmitting dengue in some localities in Sri Lanka. This investigation received financial support from University of Kelaniya (Research Grant no-97/1-23) and from the IAEA (Technical Corporation Grant no-SRL/06/024).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.