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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty

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Distribution and seasonal variation of malaria vectors in the Mannar and Jaffna Districts; an entomological investigation during the phase of prevention of re-establishment of malaria in Sri Lanka
(Springer Link, 2024) Jude, J.; Gunathilaka, N.; Wijerathna, T.; Fernando, D.; Wickremasinghe, R.; Abeyewickreme, W.
Sri Lanka has been declared “malaria-free” since 2016. The presence vectors and the cent establishment of new vectors are key challenges. This study documents the distribution of malaria vectors in previously high malaria endemic areas in Sri Lanka. Three locations were selected in Mannar and Jaffna districts. Entomological surveillance was carried out during January 2019 to January 2020 in both districts using cattle baited net (CBNT), hand collection (HC), and larval collection. The variation of mosquito abundance with selected climatic variables were evaluated using Pearson’s correlation. A total of 44,526 anophelines belonging to 12 species was encountered from both districts. The CBNT was the most productive technique that represented 78.65% (n = 35,022) of the total collection. Anopheles subpictus was the predominant species among adult collections in both districts, while in larval surveys, An. stephensi (60.1%; n = 2963) and An. varuna (40.9%; n = 1026) were observed in Jaffna and Mannar districts, respectively. The adult density indicated significantly strong positive correlations to the rainfall (r = 0.619, P < 0.05) and humidity (r = 0.524, P < 0.05) in both Jaffna and Mannar (rainfall: r = 0.553, P < 0.05; humidity: r = 0.553, P < 0.05). The number of larvae was not directly correlated with climatic variables. The present study reports the presence of An. culicifacies, An. subpictus and An. stephensi. Considering the fact that imported malaria cases are reported across the country the fact that there is an entomological risk to transmit malaria locally is a challenge. This warrants the continuous monitoring of vector population and seasonal variation, especially in previous malaria endemic regions.
Field-based evaluation of Novaluron EC10 insect growth regulator, a chitin synthesis inhibitor against Dengue vector breeding in leaf axils of pineapple plantations in Gampaha district, Sri Lanka
(BioMed Central, 2020) Gunathilaka, N.; Ranathunga, T.; Hettiarachchi, D.; Udayanga, L.; Abeyewickreme, W.
BACKGROUND: Insect growth regulators (IGRs) are considered a novel group of insecticides to control mosquitoes. Novaluron is an IGR with benzoylphenyl urea insecticide, which inhibits chitin synthesis in insects and can reduce insect population density; it is also known to have a high margin of safety for mammals. METHODS: The effective minimum concentration of novaluron formulation EC10 was tested. Six pineapple plantations [control (n = 3) and test (n = 3)] were selected from Meerigama Medical Officer of Health area in Gampaha District, Sri Lanka. Fifteen plots (10 × 10 m) were demarcated in each site with a 200 m distance apart. Leaf axils of 450 pineapple plants (30 plants × 15 plots) were screened for immature stages of Aedes mosquitoes weekly for 12 weeks. The required concentration (20 ppm) of novaluron was sprayed onto the selected pineapple plants (n = 1350) individually in 3 selected test sites for 5-10 s. The reduction in the vector population was interpreted as the percentage of reduction in immature stages of Aedes mosquitoes. RESULTS: The 100% mortality of the Ae. aegypti larvae within 24 h was observed at 20 ppm (0.05 ml of novaluron 100 g/l in 250 ml of water) as the minimum dose. Variation in the number of Aedes larvae present in the control and intervention sites was found to be significantly different throughout the entire observational period (χ2 = 128.29, df = 11, P < 0.001). The total elimination of Aedes larvae continued for up to 2 weeks and a 50% reduction was observed until the 8th week. CONCLUSIONS: The present study emphasizes that novaluron (10% EC) can be used as an effective larvicide at the treatment dose of 20 ppm. The residual effect of the IGR lasted for 12 weeks with a functional efficacy of 8 weeks. The 100% reduction of larval breeding was observed up to the 2nd week after application and the percentage reduction of immature stages remained > 50% until the 8th week. The lowest reduction (34.2%) was observed at 12 weeks after the initial treatment. Therefore, re-treatment may be recommended based on the reduction in the efficacy of the IGR. Keywords: Aedes; Field efficacy; Insect growth regulator; Novaluron; Pineapple plantation.
Determination of the efficiency of diets for larval development in mass rearing Aedes aegypti (Diptera: Culicidae)
(CABI Publishing, 2018) Gunathilaka, P.A.D.H.N.; Uduwawala, U.M.H.U.; Udayanga, N.W.B.A.L.; Ranathunge, R.M.T.B.; Amarasinghe, L.D.; Abeyewickreme, W.
Larval diet quality and rearing conditions have a direct and irreversible effect on adult traits. Therefore, the current study was carried out to optimize the larval diet for mass rearing of Aedes aegypti, for Sterile Insect Technique (SIT)-based applications in Sri Lanka. Five batches of 750 first instar larvae (L 1) of Ae. aegypti were exposed to five different concentrations (2-10%) of International Atomic Energy Agency (IAEA) recommended the larval diet. Morphological development parameters of larva, pupa, and adult were detected at 24 h intervals along with selected growth parameters. Each experiment was replicated five times. General Linear Modeling along with Pearson's correlation analysis were used for statistical treatments. Significant differences (P < 0.05) among the larvae treated with different concentrations were found using General Linear Modeling in all the stages namely: total body length and the thoracic length of larvae; cephalothoracic length and width of pupae; thoracic length, thoracic width, abdominal length and the wing length of adults; along with pupation rate and success, sex ratio, adult success, fecundity and hatching rate of Ae. aegypti. The best quality adults can be produced at larval diet concentration of 10%. However, the 8% larval diet concentration was most suitable for adult male survival.
Oviposition preferences of dengue vectors; Aedes aegypti and Aedes albopictus in Sri Lanka under laboratory settings.
(CABI Publishing, 2018) Gunathilaka, N.; Ranathunge, T.; Udayanga, L.; Wijegunawardana, A.; Abeyewickreme, W.
Investigations on oviposition behaviour of dengue vectors are critical for effective controlling of vector breeding. Therefore, the aim of the present study was to determine the oviposition behaviour of dengue vectors, Aedes aegypti and Aedes albopictus in Sri Lanka. Batches of 1000 adult mosquitoes (1 : 1, male: female ratio) housed in rearing cages were used for each experimental setup from Ae. aegypti and Ae. albopictus. Oviposition responses with respect to the size of the ovitrap, colours of the ovitrap, water source, sodium chloride (NaCl) concentration and presence/absence of larvae were evaluated by enumerating the number of eggs laid in the ovitraps. The analysis of variance and cluster analysis were used to investigate the significance in the variations among oviposition. The number of eggs laid by both species were improved with the increasing size of ovitraps. Ae. albopictus indicated the highest mean number of eggs in 0.2% of NaCl than in the ovitraps filled with distilled water. However, the egg laying preference was reduced with increasing salinity in both species. Drain water with low dissolved oxygen (DO) level (0.43 ± 0.12 mg l-1) was the preferred water source for both species, while a significantly high oviposition rate was observed in ovitraps with larvae. Black colour ovitraps attracted the majority of gravid females, while white was least preferred. There were no significant variations among oviposition behaviours of Ae. albopictus and Ae. aegypti. The ability of these vectors to breed in waste water with low DO levels may lead them to attain wide dissemination in the natural environment, enhancing their potential threat to human life.
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).
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 control
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%).
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.
Dengue vector surveillance in a dengue hot-spot in Sri Lanka
(Faculty of Tropical Medicine, Mahidol University, 2007) Sumanadasa, S.D.M.; Hapugoda, M.D.; Perera, D.; Bandara, S.; Mansoor, M.A.; Peris, I.; Abeyewickreme, W.
BACKGROUND: In South Asia, dengue has been declared as one of the most, fast-spreading vector-borne diseases. Therefore, mosquito surveillance is important for early detection of outbreaks along with implementation of prompt control activities. OBJECTIVES: To identify entomological risk factors with regard to transmission of dengue in a dengue hot-spot. Seventy five human dwellings in Vehara in the Kurunegala District of the Western Province were selected based on high disease incidence during 2000-2004, high Aedes as well as human population density and increased building activities. Entomological surveillance was done during May-August, 2007. RESULTS: The house Index ranged from 2.67% to 5.33% for Aedes aegypti while it for Aedes albopictus was 1.33% to 6.60%. The container index ranged from 23.67% to 29.33% for Ae. aegypti and from 1.33% to 18% for Ae. aibopictus. Man biting rates of 0.43-5.78 bites/man/hour were estimated for Ae, aegypti, while it ranged between 0.49 and 1.33 for Ae. aibopictus. The most common breeding place for Aedes species was plastic baskets (16%, n=12). DISCUSSIONS: Vector surveillance showed that the predominant vector species present in the study area was Ae. ageypti. Aedes mosquito larval densities and adult biting rates were sufficient to promote outbreaks of dengue in this study area. Community must be educated regarding effective measures to protect them from dengue. Their cooperation should be elicited in the early detection and elimination of vector species by source reduction, environmental management and personal protection measures.
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.

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