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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 Breeding habitat distribution of medically important mosquitoes in Kurunegala, Gampaha, Kegalle, and Kandy districts of Sri Lanka and potential risk for disease transmission: A Cross-sectional study(Hindawi Pub. Corp., 2020) Ranasinghe, K.; Gunathilaka, N.; Amarasinghe, D.; Udayanga, L.ABSTRACT: Some arbovirus infections, especially dengue, have increased rapidly over the last few decades in Sri Lanka. Prevalence and distribution of different mosquito species have been limitedly documented, which remains grossly inadequate in providing evidence for potential health risks. In this study, the diversity and species composition of mosquitoes in four selected districts in Sri Lanka (Kurunegala, Gampaha, Kegalle, and Kandy) were investigated. Entomological surveys were conducted from a total of 160 temporary and permanent mosquito breeding habitats identified in the study area from June 2017 to October 2018. Mosquito immature stages were sampled using standard dipping, siphoning, or pipetting methods and identified up to the species level. Percentage relative abundance and habitat characteristics such as species richness, dominance, and Shannon-Weiner diversity were calculated for each surveyed habitat type. Associations between co-occurring species were estimated by Hulbert's coefficient of interspecific association (C8). A total of 4663 mosquito larvae belonging to seven genera and fifteen species of mosquitoes were collected. The relative distribution of mosquito species differed significantly among the four studied districts (X 2 = 143.248; df = 33; P < 0.001). According to Kruskal-Wallis statistics (P < 0.05 at 95% of significance), all diversity indices for immature stages of medically important mosquitoes varied significantly across different breeding sites. Paddy fields had the significantly highest species richness of 4.0 ± 2.82. The coefficients of interspecific association among all the recorded medically important vector mosquitoes were found negative during the present study. The findings of the current study would be useful to identify the entomological potential for disease transmission and facilitate the implementation of appropriate vector control interventions. This would ultimately provide an avenue to improve the personal skills of health staff rather than limiting their knowledge to specified disease vectors, under which the control program is concerned.Item Aedes aegypti lines for combined sterile insect technique and incompatible insect technique applications: the importance of host genomic background(Blackwell Publishing Ltd, 2020) Carvalho, D.O.; Torres-Monzon, J.A.; Koskinioti, P.; Wijegunawardana, N.D.A.D.; Liang, X.; Pillwax, G.; Xi, Z.; Bourtzis, K.ABSTRACT: Aedes aegypti L. (Diptera: Culicidae), being the primary vector of pathogenic arboviruses, is a target for the development of novel genetic approaches to complement current conventional vector control strategies such as the combined sterile insect and incompatible insect technique (SIT/IIT). A transinfected line of Ae. aegypti carrying the wAlbB Wolbachia strain (WB2) was introgressed into two genomic backgrounds, Brazil and Mexico, producing two new Ae. aegypti strains (WB2-BRA and WB2-MEX). These strains were evaluated with respect to several life-history traits such as fecundity, fertility, longevity, pupa size, pupation curve, and male mating competitiveness, as well as their response to irradiation. Our results show that the impact of Wolbachia infection depends on the genomic background and that the Brazilian one had no significant effect, whereas the Mexican one negatively affected fertility, longevity, and pupal size. Interestingly, Wolbachia-infected Ae. aegypti lines required a lower irradiation dose to achieve complete female sterility than the uninfected ones. The present findings are discussed given the potential use of Wolbachia-infected Ae. aegypti lines in combined SIT/IIT population suppression programs.Item Larval indices of vector mosquitoes as predictors of dengue epidemics: An approach to manage dengue outbreaks based on entomological parameters in the districts of Colombo and Kandy, Sri Lanka(Hindawi Pub. Co., 2020) Udayanga, L.; Aryaprema, S.; Gunathilaka, N.; Iqbal, M. C. M.; Fernando, T.; Abeyewickreme, W.BACKGROUND: Early detection of dengue epidemics is a vital aspect in control programmes. Predictions based on larval indices of disease vectors are widely used in dengue control, with defined threshold values. However, there is no set threshold in Sri Lanka at the national or regional levels for Aedes larval indices. Therefore, the current study aimed at developing threshold values for vector indices in two dengue high-risk districts in Sri Lanka. METHODS: Monthly vector indices (House Index [HI], Container Index [CI], Breteau Index for Aedes aegypti [BIagp], and Ae. albopictus [BIalb]), of ten selected dengue high-risk Medical Officer of Health (MOH) areas located in Colombo and Kandy districts, were collected from January 2010 to June 2019, along with monthly reported dengue cases. Receiver Operating Characteristic (ROC) curve analysis in SPSS (version 23) was used to assess the discriminative power of the larval indices in identifying dengue epidemics and to develop thresholds for the dengue epidemic management. RESULTS: Only HI and BIagp denoted significant associations with dengue epidemics at lag periods of one and two months. Based on Ae. aegypti, average threshold values were defined for Colombo as Low Risk (2.4 ≤ BIagp < 3.8), Moderate Risk (3.8 ≤ BIagp < 5), High Risk (BIagp ≥ 5), along with BIagp 2.9 ≤ BIagp < 4.2 (Low Risk), 4.2 ≤ BIagp < 5.3 (Moderate Risk), and BIagp ≥ 5.3 (High Risk) for Kandy. Further, 5.5 ≤ HI < 8.9, 8.9 ≤ HI < 11.9, and HI ≥ 11.9 were defined as Low Risk, Moderate Risk, and High Risk average thresholds for HI in Colombo, while 6.9 ≤ HI < 9.1 (Low Risk), 8.9 ≥ HI < 11.8 (Moderate Risk), and HI ≥ 11.8 (High Risk) were defined for Kandy. CONCLUSIONS: The defined threshold values for Ae. aegypti and HI could be recommended as indicators for early detection of dengue epidemics and to drive vector management activities, with the objective of managing dengue epidemics with optimal usage of financial, technical, and human resources in Sri Lanka.Item Gene flow patterns among Aedes aegypti (Diptera: Culicidae) populations in Sri Lanka(MDPI AG, 2020) Fernando, H.S.D.; Hapugoda, M.; Perera, R.; Black Iv, W.C.; de Silva, B.G.D.N.K.ABSTRACT:In Sri Lanka, dengue is the most serious arboviral disease. Recent increases in dengue cases suggest a higher infection rate and spread of the disease to new areas. The present study explores gene flow patterns of Ae. aegypti, the main vector of dengue disease, among 10 collection sites including major ports and inland cities using variations at 11 microsatellite loci. Discriminant analysis of principal components (DAPC) and k-means clustering estimated eight genetic clusters. Analysis of Molecular Variance (AMOVA) estimated equal variances among cities and among collections in Colombo, Sri Lanka. Significant evidence, although weak, was detected for isolation by distance. Analysis of gene flow rates and directions using MIGRATE-n indicated that populations throughout the island served as a source of immigrants for Colombo with abundant gene flow among major commercial cities in Sri Lanka, which appear to receive migrant mosquitoes from throughout Sri Lanka. The observed patterns probably arise through human movement of Ae. aegypti during commerce from throughout Sri Lanka into Colombo increasing the risk of spread. The patterns uncovered in this study are significant for global health as Sri Lanka is situated along a key international shipping route. KEYWORDS: Aedes aegypti; Sri Lanka; gene flow patterns; population structure.Item Use of mechanical and behavioural methods to eliminate female Aedes aegypti and Aedes albopictus for sterile insect technique and incompatible insect technique applications.(BioMed Central, 2019) Gunathilaka, N.; Ranathunge, T.; Udayanga, L.; Wijegunawardana, A.; Gilles, J. R. L.; Abeyewickreme, W.BACKGROUND:Sex separation of mosquitoes at different stages is currently being attempted to ensure the successful release of male mosquitoes in novel vector control approaches. Mechanical and behavioral techniques have been tried most frequently. METHODS: Batches of (n = 300) Aedes aegypti and Ae. albopictus pupae were used for standard sieving (using sieves with 1.12, 1.25, 1.40 and 1.60 mm mesh sizes) and the Fay-Morlan glass plate separation methods. Male and female separation by each method was calculated. For behavioral separation, a spiked blood meal with different concentrations (0, 2, 4, 6, 8 and 10 ppm) of ivermectin and spinosad (spinosyn, 12% w/v), were provided to a batch (n = 300) of adult Ae. aegypti and Ae. albopictus (1:1 sex ratio) followed by observation of mortality. An additional "double feeding method" involved provision of a further blood meal after 24 h, with the same concentrations of ivermectin and spinosad as the initial feeding, followed by a 48-h observation of mortality. All experiments were repeated five times. RESULTS: In the standard sieving method, the percentage of males and females separated at different pore sizes differed significantly (P < 0.05). The majority of the male pupae were collected in the 1.12 mm pore sized sieve for both Ae. aegypti (73%) and Ae. albopictus (69%) while females were retained mainly in the sieve with the pore size of 1.25 mm. In the Fay-Morlan glass plate separation, 99.0% of the Ae. aegypti and 99.2% of the Ae. albopictus introduced male pupae could be separated, but with female contaminations of 16 and 12%, respectively. Provision of a blood meal spiked with 8 ppm of ivermectin under the "double feeding" was identified as the most effective way of achieving 100% female elimination for both Aedes species. CONCLUSIONS: With 100% separation, use of a spiked blood meal is a more effective method of sex separation than the mechanical methods. Application of the spiked blood meal approach as a second separation level for sexes, after applying the Fay-Morlan glass plate method, could achieve 100% sex separation of sexes whilst allowing a reduction in the amount of toxicants required.Item First report of V1016G and S989P knockdown resistant (kdr) mutations in pyrethroid-resistant Sri Lankan Aedes aegypti mosquitoes(BioMed Central, 2018) Fernando, S.D.; Hapugoda, M.; Perera, R.; Saavedra-Rodriguez, K.; Black WC 4th; de Silva, N.K.; ; ;BACKGROUND: Dengue is a serious arboviral disease in Sri Lanka with a large number of dengue fever (DF) cases every year. Control of the primary vector Aedes aegypti depends upon larval habitat source reduction and insecticide application. However, increases in the number of reported cases suggest the inefficiency of current control strategies and the possibility of resistance to currently used insecticides. Early detection of mutations in the voltage-gated sodium channel (vgsc) gene that confer knockdown resistance (kdr) to pyrethroid insecticides is important in resistance management in vector populations. RESULTS: Resistance to pyrethroid insecticides was detected in the three populations studied. Polymerase chain reaction was used to detect the presence of two kdr mutations F1534C and V1016G. During this process a S989P mutation was also detected in pyrethroid-resistant Ae. aegypti populations. These mutations were found to be widespread and frequent in the collections studied. CONCLUSIONS: To our knowledge, this study reveals for the first time the presence of V1016G and S989P mutant alleles in the vgsc of Sri Lankan Ae. aegypti populations. The spread of the mutant alleles throughout the country poses a threat of increased resistance to pyrethroids. Long-term insecticide applications and indiscriminate use of pyrethroids has led to the evolution of resistance. More strategic and diverse strategies, including novel insecticides with new modes of action and community participation, should be engaged for Ae. aegypti control.Item A Statistical Approach to Define Thresholds for Dengue Epidemic Management in Akurana Medical Officer of Health Area, Kandy District of Sri Lanka(19th Conference on Postgraduate Research, International Postgraduate Research Conference 2018, Faculty of Graduate Studies,University of Kelaniya, Sri Lanka, 2018) Udayanga, N.W.B.A.L.; Gunathilaka, P.A.D.H.N.; Iqbal, M.C.M.; Fernando, M.A.S.T.; Abeyewickreme, W.Stegomyia indices, namely; Premise Index (PI), Breteau Index (BI) and Container Index (CI) are used forvector management approaches in Sri Lanka. Properly defined threshold values for larval indices are of higher importance to provide forecasts on dengue epidemics and also for effective larval management of dengue vectors. However, such critical thresholds are poorly defined for Sri Lanka. The present study aimed to define threshold values forabove larval indices for dengue epidemic management in the Akurana Medical Officer of Health (MOH) in the Kandy District. Larval surveys were conducted on a monthly basis from January, 2016 to June, 2018. Four larval indices, namely BI for Aedesaegypti (BIA) and Aedesalbopictus (BIB), PI and CI were calculated. Further, monthly larval indices of AkuranaMOH area from January, 2012 to December, 2015, were obtained from the MOH office, along with monthly reported dengue cases for the entire study period. Receiver Operating Characteristic (ROC) curves in SPSS (version 23) were used to assess the discriminative power of the larval indices in determiningdengue epidemics and thresholds based on larval indices. As indicated by the area of ROC curve (AUC), the BIA (0.661) and PI (0.637) were having a notable discriminative power to forecast dengue epidemics at a two-month lag period. Both BIB (0.397) and CI (0.526) were non-informative influencers at one and two-month lag periods. The BIA and PI were better predictors of dengue incidence than BIB and CI. Based on the ROC curve, three risk thresholds were defined for BIA as Low Risk (BIA≤2.1), Moderate Risk (3.9≤BIA<4.85), and High Risk (BIA≥4.85), with respect to Ae. aegypti. According to the PI, thresholds were defined as Low Risk (PI≤6.2), Moderate Risk (7.7≤ PI<9.9), and High Risk (PI≥ 9.9). Threshold values defined for BI of Ae. aegypti and PI, could be recommended to be considered in implementing vector control efforts in the above study area for effective dengue epidemic management, through pre planned entomological management of dengue vectors.Item Empirical optimization of risk thresholds for dengue: an approach towards entomological management of Aedes mosquitoes based on larval indices in the Kandy District of Sri Lanka(BioMed Central, 2018) Udayanga, L.; Gunathilaka, N.; Iqbal, M.C.M.; Najim, M.M.M.; Pahalagedara, K.; Abeyewickreme, W.BACKGROUND: Larval indices such as Premise Index (PI), Breteau Index (BI) and Container Index (CI) are widely used to interpret the density of dengue vectors in surveillance programmes. These indices may be useful for forecasting disease outbreaks in an area. However, use of the values of these indices as alarm signals is rarely considered in control programmes. Therefore, the current study aims to propose threshold values for vector indices based on an empirical modeling approach for the Kandy District of Sri Lanka. METHODS: Monthly vector indices, viz PI, BI and CI, for Aedes aegypti and Aedes albopictus, of four selected dengue high risk Medical Officer of Health (MOH) areas in the Kandy District from January 2010 to August 2017, were used in the study. Gumbel frequency analysis was used to calculate the exceedance probability of quantitative values for each individual larval index within the relevant MOH area, individually and to set up the threshold values for the entomological management of dengue vectors. RESULTS: Among the study MOH areas, Akurana indicated a relatively high density of both Ae. aegypti and Ae. albopictus, while Gangawata Korale MOH area had the lowest. Based on Ae. aegypti, threshold values were defined for Kandy as low risk (BIagp > 1.77), risk (BIagp > 3.23), moderate risk (BIagp > 4.47) and high risk (BIagp > 6.23). In addition, PI > 6.75 was defined as low risk, while PI > 9.43 and PI>12.82 were defined as moderate and high risk, respectively as an average. CONCLUSIONS: Threshold values recommended for Ae. aegypti (primary vector for dengue) along with cut-off values for PI (for Ae. aegypti and Ae. albopictus), could be suggested as indicators for decision making in vector control efforts. This may also facilitate the rational use of financial allocations, technical and human resources for vector control approaches in Sri Lanka in a fruitful manner.Item Establishment and maintenance of laboratory colonies of Aedes albopictus mosquitoes(University of Peradeniya, 2015) Wijegunawardana, A.D.; Gunathilaka, H.N.; Dassanayake, R.; Gunawardene, Y.I.N.S.; Abeyewickreme, W.With a mission of "providing authenticated, high-quality Aedes albopictus mosquito rearing information to the research community" maintenance of a Ae. albopictus mosquito colony was started. All environmental facilities inside the insectary were carefully maintained to better suit the Ae. albopictus mosquito colonization. The mean temperature of 27°C (± 0.5°C) was constantly maintained inside the insectary. Wet towels on adult mosquito cage racks were used for proper maintenance of humidity. Lighting was using fluorescent light and regulated with 16:8 hour continuous dark and light period. Pest insect was controlled to ensure essential absence of ants and cockroaches. This was achieved without any harm to the mosquito colonies either directly or by contamination with toxicants transported by pests. An adult mosquito trap placed inside the insectary was used to monitor released mosquitoes. Consistent effort was also made to improve the level of cleanliness inside the insectary. Written guidelines were given to each person responsible for a task. Insectary operations included egg counting, preparation of hatching bottles with boiled distilled water following cooling to room temperature, egg hatching, larvae rearing with International Atomic Energy Agency (IAEA) recommended diet of tuna meal, bovine liver powder, brewery yeast and vitamin complex in a ratio of 37.5:27:10.5:2 g in 1L up to one week, pupae counting and putting into adult emergency cages, adult male feeding with 10% sugar solution with Vitamin complex, adult female blood feeding from 4th day onwards with bovine blood, placing egg laying cups and collecting egg laying cups, drying egg papers and starting next generation from the dried eggs. Adult mosquito cages were blood fed every 4th day after emergence from pupa and for quality control reasons each adult cage was blood fed only 3 times and there after only 10% sugar solution with vitamin syrup was supplemented until all adult mosquitoes died. Documentation for maintenance and data record was maintained and updated daily. Records included larvae feeding records, larvae tray maintenance and cleaning charts, adult feeding records with both sugar solution and blood, insectary cleaning records with time and dates. Number of eggs and percentage of egg hatching, larvae death, pupation, adult emergence, egg laying and adult mosquito death with respect of the sex and time difference were recorded. For bio-safety reasons all discarded material from larvae trays, egg laying cups and adult cages were boiled thoroughly to facilitate total destruction of the contaminated mosquito eggs. All other infectious material were incinerated. Finally, all above conditions facilitated achievement of 100% egg hatching rate within maximum of 24 hours, 100% survival of larvae to pupa (~ 7 days), 100% survival of pupated larvae to adult emergence (~ 2 days) and 95.5% adult survival up to 12 days. No difference was observed on adult longevity between males and females within first 12 days of adult emergence. However, approximate life span for males (-17 days) was lower than the females (~ 25 days) and the mortality was regular through all generations (Fl to F21).Item 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