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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 Forecasting dengue incidence based on entomological indices, population density, and meteorological and environmental variables in the Gampaha District of Sri Lanka(Elsevier Ltd, 2024) Dalpadado, R.; Amarasinghe, D.; Gunathilaka, N.; Wijayanayake, A.N.No abstract availableItem Bionomic aspects of dengue vectors Aedes aegypti and Aedes albopictus at domestic settings in urban, suburban and rural areas in Gampaha District, Western Province of Sri Lanka(BioMed Central, London, 2022) Dalpadado, R.; Amarasinghe, D.; Gunathilaka, N.; Ariyarathna, N.Background: The lack of information on behavioural patterns of Aedes aegypti and Aedes albopictus has become a significant limitation in vector control and disease management programmes. Therefore, the current study was focused on determining some bionomics aspects: breeding, resting, host-seeking and feeding preferences of Ae. aegypti and Ae. albopictus in Sri Lanka. Methods: Larval and adult surveys were conducted from April 2017 to April 2019 monthly in six selected Medical Officer of Health (MOH) areas in Gampaha Distinct, Western province, Sri Lanka, representing urban, suburban and rural settings. Resting preferences of adult mosquitoes were observed from indoor and outdoor places using a Prockopack aspirator. The information on resting height, surface, material and locality was recorded. Human-baited double-net traps were used to determine the host-seeking time of Aedes mosquitoes. Statistical differences in the spatial distribution of mosquitoes in selected MOH areas and prevalence of vectors were analysed using general linear model (GLM). A chi-square test was used to analyse the resting behaviour. Results: Total of 19,835 potential breeding sites were examined at 13,563 premises, and 18.5% (n = 1856) were positive for Aedes larvae. Distribution of Ae. aegypti and Ae. albopictus was statistically significant at species level (df = 1; F = 137.134; P < 0.05 GLM) and study setting (df = 2; F = 8.125; P < 0.05). Aedes aegypti breeding was found mainly in temporary removals (18.8%; n = 34), discarded non-degradables (12.15%; n = 22) and tyres (9.95%; n = 18). Natural (14.7%; n = 246) and temporary removals (13.6%; n = 227) and discarded non-reusable items were the key ovipositing sites for Ae. albopictus. In the adult mosquito survey, the majority was comprised of Ae. albopictus (54.5%; n = 999), which denoted exophilic nature (90.8%; n = 758), and 45.5% (n = 835) represented by Ae. aegypti mosquitoes who were mainly endophilic (84.3%; n = 842). Aedes aegypti rested on cloth hangings and curtains, followed by the furniture, while Aedes albopictus was predominant in outdoor vegetation. In both vectors, biting patterns denoted a typical diurnal pattern with two peaks of host-seeking and biting activity in the morning and afternoon. Conclusions: The majority (80%) of the larval habitats were artificial containers. The use of larvicides for vector control as the prominent measure is questionable since applying these chemicals may target only 20% of the total breeding grounds, which are permanent. The resting places of adult mosquitoes are mainly indoors. Therefore, using thermal space spraying of insecticide may not be appropriate, and indoor residual spraying is recommended as a suitable intervention to target adult mosquitoes. This study warrants a holistic vector control approach for all medically important mosquitoes and insects, ensuring the rational use of finance and resources.Item Water quality characteristics of breeding habitats in relation to the density of Aedes aegypti and Aedes albopictus in domestic settings in Gampaha district of Sri Lanka(Elsevier, 2022) Dalpadado, R.; Amarasinghe, D.; Gunathilaka, N.In spite of the unavailability of measures to target dengue control in human populations, the promising way of combating the disease is by controlling vector mosquito larvae and their breeding habitats. Water quality characteristics of the breeding habitats of dengue vectors are among the crucial parameters that determine the female mosquito's oviposition and breeding. Therefore, a cross-sectional study was conducted to investigate the habitat characteristics of dengue vector mosquitoes by assessing the water quality of positive breeding habitats using the weighted arithmetic water quality index (WQI). The present study was conducted in domestic areas of the selected medical officer of health areas of the Gampaha district, Sri Lanka, from 2017 to 2019. Water quality characteristics of breeding habituations were measured and analyzed. The relationship between water quality parameters in breeding habitats with the occurrence of mosquito species was assessed using Kruskal-Wallis one-way analysis of variance, followed by pairwise comparison using Dunn's test and Mann-Whitney U test at a 5% level of significance. This study revealed that the temperature of Aedes mosquito breeding water ranged between 25.3 and 39.8 °C, and bred at temperatures as high as 39.8 °C in discarded receptacles. The results indicated that Ae. aegypti was prominent in alkaline water ranging between 7.5 and 8.5 pH, whereas Ae. albopictus was abundant in water with a pH range of 6.5-7.5. Both species of Aedes inhabited waters of low turbidity and TDS level. Ae. aegypti and Ae. albopictus immatures were prominent in water where TDS levels ranged between 250 and 350 ppm. The mean conductivity in the mosquito breeding water was recorded as 228.3 ± 63.9 µs/cm. The study revealed that Aedes mosquitoes could breed in water with a mean dissolved oxygen level of 6.9 ± 0.7 mg/L, ranging between 6.35 ± 1.09 mg/L and 7.28 ± 0.26 mg/L. The water quality indices were calculated for the eight previously identified breeding habitat categories of Aedes mosquitoes.Item A Systematic review on diagnostic methods of red cell membrane disorders in Asia(Blackwell Scientific Publications, 2022) Silva, R.; Amarasinghe, D.; Perera, S.; Premawardhena, A.Membranopathies are a group of inherited blood disorders where the diagnosis could form a challenge due to phenotype-genotype heterogeneity. In this review, the usage and limitations of diagnostic methods for membranopathies in Asian countries were evaluated. A systematic review was done using articles from PubMed, Google Scholar, and EBSCO from 2000 to 2020. Thirty-six studies conducted in seven Asian countries had used different diagnostic methods to confirm membranopathies. In 58.3% of studies, full blood count (FBC), reticulocyte count, and peripheral blood smear (PBS) were used in preliminary diagnosis. The combination of the above three with osmotic fragility (OF) test was used in 38.8%. The flowcytometric osmotic fragility (FC-OF) test was used in 27.7% where it showed high sensitivity (92%-100%) and specificity (96%-98%). The eosin-5-maleimide (EMA) assay was used in 68.1% with high sensitivity (95%-100%) and specificity (93%-99.6%). About 36.1% of studies had used sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a further diagnostic method to detect defective proteins. Genetic analysis to identify mutations was done using Sanger sequencing, next-generation sequencing (NGS), and whole-exome sequencing (WES) in 33.3%, 22.2%, and 13.8% of studies, respectively. The diagnostic yield of NGS ranged from 63% to 100%. Proteomics was used in 5.5% of studies to support the diagnosis of membranopathies. A single method could not diagnose all membranopathies. Next-generation sequencing, Sanger sequencing, and proteomics will supplement the well-established screening and confirmatory methods, but not replace them in hereditary hemolytic anemia assessment.Item Diversity of midgut bacteria in larvae and females of Aedes aegypti and Aedes albopictus from Gampaha District, Sri Lanka(BioMed Central, 2021) Ranasinghe, K.; Gunathilaka, N.; Amarasinghe, D.; Rodrigo, W.; Udayanga, L.BACKGROUND: The midgut microbiota of mosquitoes maintain basal immune activity and immune priming. In recent years, scientists have focused on the use of microbial communities for vector control interventions. In the present study, the midgut bacteria of larvae and adults of Aedes aegypti and Ae. albopictus were assessed using both field-collected and laboratory-reared mosquitoes from Sri Lanka. METHODS: Adults and larvae of Ae. aegypti and Ae. albopictus were collected from three selected areas in Gampaha Medical Officer of Health area, Gampaha District, Western Province, Sri Lanka. Bacterial colonies isolated from mosquito midgut dissections were identified by PCR amplification and sequencing of partial 16S rRNA gene fragments. RESULTS: Adults and larvae of Ae. aegypti and Ae. albopictus harbored 25 bacterial species. Bacillus endophyticus and Pantoea dispersa were found more frequently in field-collected Ae. aegypti and Ae. albopictus adults, respectively. The midgut bacteria of Ae. aegypti and Ae. albopictus adults (X2 = 556.167, df = 72, P < 0.001) and larvae (X2 = 633.11, df = 66, P < 0.001) were significantly different. There was a significant difference among the bacterial communities between field-collected adults (X2 = 48.974, df = 10, P < 0.001) and larvae (X2 = 84.981, df = 10, P < 0.001). Lysinibacillus sphaericus was a common species in adults and larvae of laboratory-reared Ae. aegypti. Only P. dispersa occurred in the field-collected adults of Ae. aegypti and Ae. albopictus. Species belonging to genera Terribacillus, Lysinibacillus, Agromyces and Kocuria were recorded from Aedes mosquitoes, in accordance with previously reported results. CONCLUSIONS: This study generated a comprehensive database on the culturable bacterial community found in the midgut of field-collected (Ae. aegypti and Ae. albopictus) and laboratory-reared (Ae. aegypti) mosquito larvae and adults from Sri Lanka. Data confirm that the midgut bacterial diversity in the studied mosquitoes varies according to species, developmental stage and strain (field vs laboratory). KEYWORDS: 16S rRNA gene; Aedes; Bacteria; Diversity; Midgut; Mosquitoes.Item A Challenge for a unique dengue vector control programme: Assessment of the spatial variation of Insecticide resistance status amongst Aedes aegypti and Aedes albopictus populations in Gampaha District, Sri Lanka(BioMed Central, 2021) Dalpadado, R.; Gunathilaka, N.; Amarasinghe, D.; Udayanaga, L.BACKGROUND: To date, dengue is considered an important public health problem in Sri Lanka. Irrational use of insecticides without evidence-based applications has primed the development of resistance in mosquito vectors. METHOD: The present study investigated the resistance status of Aedes aegypti and Aedes albopictus to commonly used insecticides in three selected Medical Officer of Health (MOH) areas (i.e., Attanagalla, Dompe, and Negombo) in Gampaha District, Western Province of Sri Lanka. Entomological surveys were performed using ovitraps and larval collections. Larval bioassays were carried out to determine the LC50, LC90, and LC95 and susceptibility status for organophosphate temephos, whereas adult bioassays were performed to test the 0.03% deltamethrin and 0.8% malathion susceptibility. RESULTS: The study revealed that the temephos concentrations required to control Ae. aegypti (13.7-17.7 times) and Ae. albopictus (4.6-7.6 times) are higher than the diagnostic concentration (0.012 mg/L) proposed by the World Health Organization. The highest resistance levels were observed for both Ae. aegypti (14 ± 1.87) and Ae. albopictus (36 ± 1.87) collected from the Negombo MOH area. Therefore, the WHO recommended diagnostic concentration is no longer effective in controlling Ae. aegypti and Ae. albopictus larvae in these areas. Both the dengue vectors have evolved a high level of insecticide resistance to malathion and deltamethrin in the Gampaha District except Ae. albopictus mosquitoes in rural areas. Further, vectors in rural areas are indicated susceptible (>98%) to pyrethroids and emergence of resistance (<97%) for organophosphate insecticides. CONCLUSION: The results of this study warrant the vector management authorities on the proper application of insecticides and rational use in vector control. The susceptibility status of vector mosquitoes should be continuously monitored especially in dengue-endemic areas parallel to the routine surveillance programme. Further molecular studies are strongly recommended to determine the Knockdown Resistance (kdr) mutations among Aedes populations.Item Molecular characterization of culturable aerobic bacteria in the midgut of field-caught Culex tritaeniorhynchus, Culex gelidus, and Mansonia annulifera mosquitoes in the Gampaha district of Sri Lanka(Hindawi Pub. Co., 2020) Gunathilaka, N.; Ranasinghe, K.; Amarasinghe, D.; Rodrigo, W.; Mallawarachchi, H.; Chandrasena, N.Background: Larval and adult mosquito stages harbor different extracellular microbes exhibiting various functions in their digestive tract including host-parasite interactions. Midgut symbiotic bacteria can be genetically exploited to express molecules within the vectors, altering vector competency and potential for disease transmission. Therefore, identification of mosquito gut inhabiting microbiota is of ample importance before developing novel vector control strategies that involve modification of vectors. Method: Adult mosquitoes of Culex tritaeniorhynchus, Culex gelidus, and Mansonia annulifera were collected from selected Medical Officer of Health (MOH) areas in the Gampaha district of Sri Lanka. Midgut lysates of the field-caught non-blood-fed female mosquitoes were cultured in Plate Count Agar medium, and Prokaryotic 16S ribosomal RNA partial genes of the isolated bacteria colonies were amplified followed by DNA sequencing. Diversity indices were used to assess the diversity and richness of the bacterial isolates in three mosquito species. The distribution pattern of bacterial isolates between different mosquito species was assessed by Distance-Based Redundancy Analysis (dbRDA). Results: A total of 20 bacterial species (Staphylococcus pasteuri, Bacillus megaterium, Staphylococcus cohnii, Pantoea dispersa, Staphylococcus chromogenes, Bacillus aquimaris, Staphylococcus arlettae, Staphylococcus sciuri, Staphylococcus warneri, Moraxella osloensis, Enterobacter sp., Klebsiella michiganensis, Staphylococcus hominis, Staphylococcus saprophyticus, Streptomyces sp., Bacillus niacin, Cedecea neteri, Micrococcus luteus, Lysinibacillus sphaericus, and Bacillus licheniformis) were identified. All of these species belonged to three phyla, Proteobacteria, Firmicutes, and Actinobacteria, out of which phylum Firmicutes (71.1%) was the most prominent. The least number of species was recorded from Actinobacteria. The relative distribution of midgut microbes in different mosquito species differed significantly among mosquito species (Chi-square, χ 2 = 486.091; df = 36; P ≤ 0.001). Midgut microbiota of Cx. tritaeniorhynchus and Cx. gelidus indicated a similarity of 21.51%, while Ma. annulifera shared a similarity of 6.92% with the cluster of above two species. The gut microbiota of Cx. tritaeniorhynchus was also significantly more diverse and more evenly distributed compared to Ma. annulifera. Simpson's diversity, Margalef's diversity, and Menhinick's diversity indices were higher in Cx. gelidus. Of the recorded species, P. dispersa and strains of nonpathogenic species in Bacillaceaefamily (B. megaterium, B. niacini, B. licheniformis, and L. sphaericus) can be recommended as potential candidates for paratransgenesis. Conclusion: The relative distribution of midgut microbes in different mosquito species differed significantly among the three studied adult mosquito species. The present data strongly encourage further investigations to explore the potential usage of these microbes through paratransgenic approach for novel eco-friendly vector control strategies.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 Effect of larval nutritional regimes on morphometry and vectorial capacity of Aedes aegypti for Dengue transmission(BioMed Research International, 2019) Gunathilaka, N.; Upulika, H.; Udayanga, L.; Amarasinghe, D.BACKGROUND: Nutritional level in larval diet of mosquito vectors influence on life history traits and vectorial capacity (VC). Therefore, the present study was carried out to assess the effect of larval diet concentration on vector bionomic and VC of Aedes aegypti in Sri Lanka. Method. Three batches of 400 Ae. aegypti larvae (first instar) were reared under different concentrations of larval diet (6%, 8%, and 10%; Volume/Volume), which was prepared by mixing 12.5 g of tuna meal, 9.0 g of bovine liver powder, and 3.5 g of Brewer’s yeast, in 100 ml of distilled water. The effect of larval diet concentration on different morphometric and functional parameters of larvae (length and width of head, abdomen, survival rate, and pupation success), pupae (length and width of cephalothorax, survival rate, and adult emersion), adult (length and width of thorax, abdomen, survival rate, longevity, biting frequency and fecundity of adults) were examined. In addition, VC of Ae. aegypti was evaluated. The General Linear Model (GLM) was used for the statistical analysis. Results. Larval head length, head width, thoracic width, abdominal length, abdominal width, total length, and survival rate significantly increased with higher doses of larval diet (P<0.05). In case of pupae, length, and width of cephalothorax, survival rate and adult emergence rate denoted an increasing trend with the elevated larval diets. However, the variations of survival rate and adult emergence rate were statistically significant (P<0.05). In adults, all morphometric parameters (thoracic length, abdominal length, abdominal width, and wing length) significantly increased with elevating larval diets levels (except for thoracic width) along with the biting frequency, fecundity, and survival rate (P<0.05) of adult females. The VC also denoted significant variations (F4,14 = 24.048; ) with the larval diet concentration, whereby the highest VC of 196.37 was observed at 10% treatment. Conclusion. Larval food availability has a significant influence on the adult fitness and thus may affect the incidence of dengue due to variations in the VC of Ae. aegypti. Hence, this investigation highlights the requirement in accounting the environmental variation at the larval stages in order to understand transmission dynamics and control of dengue in Sri Lanka.Item Prevalence of gastrointestinal parasitic infections and assessment of deworming program among cattle and buffaloes in Gampaha District, Sri Lanka(Hindawi Publishers, 2018) Gunathilaka, N.; Niroshana, D.; Amarasinghe, D.; Udayanga, L.Gastrointestinal (GI) parasitic infection is a serious issue in cattle management. The effects of GI parasites may vary with age, sex of cattle, nutritional condition, and severity of infection. Prevalence of GI parasites among cattle population in Gampaha District has not been studied and there is no published study available. A total of 45 farms rearing cattle were selected randomly in three areas, namely, Kelaniya, Ganemulla, and Welisara, under three Veterinary Surgeon Divisions (VSD) in Gampaha District (Mahara, Gampaha, and Welisara). Freshly voided cattle fecal samples were collected randomly from the selected farms during March 2017–December 2017. Out of 163 cattle and buffaloes examined, 13.39% (n=22) were positive for eggs of one or more species of GI parasites. The prevalence of parasitic infection was higher in buffaloes (31.25%, 5/16) as compared to that of cows (11.56%, 21/147), but the difference was not significant (P >0.05). Hookworms (Bunostomum spp.), whipworms (Trichuris spp.), digenetic trematodes (Paramphistomum spp.), cestodes (Moniezia spp.), and oocysts of protozoans (coccidians) were found during the study. The nontreated animals indicated the highest percentage of parasitic infections accounting for 46.67% (n= 14), followed by partially treated individuals (15.15%, n= 5). GI parasite prevalence in males was higher when compared to that of females, but the difference was nonsignificant (P >0.05). General Linear Modelling (GLM) revealed that the effect of treatment status was significantly associated with the prevalence of GI parasites. The calves and yearlings had the highest rate of GI parasitic infections. The highest infection rate was observed at Kelaniya, followed by Welisara. Future investigations are necessary to evaluate the economic impact of GI parasites in the study areas.