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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 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.Item Ocular trematodiasis in children, Sri Lanka(National Center for Infectious Diseases, 2023) Mallawarachchi, C.H.; Dissanayake, M.M.; Hendavitharana, S.R.; Senanayake, S.; Gunathilaka, N.; Chandrasena, T.G.A.N.; Yahathugoda, T.C.; Wickramasinghe, S.; de Silva, N.R.Using histopathology and phylogenetic analysis of the internal transcribed spacer 2 gene, we found >2 distinct trematode species that caused ocular trematode infections in children in Sri Lanka. Collaborations between clinicians and parasitologists and community awareness of water-related contamination hazards will promote diagnosis, control, and prevention of ocular trematode infections.Item Comparative analysis of the larvicidal activity of temephos (EC50) and novaluaron (EC10) to control Anopheles stephensi in Sri Lanka(Elsevier, 2023) Jude, J.; Gunathilaka, N.; Udayanga, L.; Fernando, D.; Premarathne, P.; Wickremasinghe, R.; Abeyewickreme, W.Background: Anopheles stephensi was first recorded in the coastal area of Mannar District, Sri Lanka, in December 2016. Since then, this vector has been isolated from other districts in the Northern and Eastern Provinces of Sri Lanka. Chemical control is the main arm of vector control that can be used to reduce the vector densities within a short period. Thus, the present study aimed at evaluating the efficacy of using selected insecticides for the control of An. stephensi larvae. Method: The third and fourth instar larval stages of An. stephensi (F2 generation) of field mosquitoes that were caught using cattle baited net trap collections from Columbuthurai, Kurunagar, and Navanthurai areas in Jaffna District, Sri Lanka, were obtained from the laboratory colony established at Jaffna. Batches of 100 larvae were taken for experiments and introduced separately to a concentration series of temephos and novaluron (0.04-400 ppm). A control test was also performed at each setup without introducing insecticides. The mortality rates of An. stephensi larvae exposed to different concentrations of larvicides were recorded at 1, 24 and 48-h intervals. The experiment was replicated five times at individual concentrations for each selected chemical. Data were analyzed using the General Linear Model (GLM) and Probit analysis. Results: The highest mortality rate (100%) at a 1-h exposure period was observed from temephos at >100 ppm. The mortality rates varied significantly for different concentrations and larvicides (p < 0.05). At 24-h of the exposure period, the 100% mortality of An. stephensi larvae were observed from both temephos and novaluron even at 0.04 ppm. Conclusion: Both temephos and novaluron reported 100% mortality rates in An. stephensi larvae at 1-h and 24-h exposure periods. Based on the findings, temephos and novaluron can be recommended as effective larvicides for chemical-based control of An. stephensi in Jaffna, Sri Lanka. Further, it is recommended to conduct a field-based study, where habitat types and water quality are highly heterogeneous and may affect the residual activity.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 Genetic diversity of Leishmania donovani isolates from cutaneous lesions of military personnel in the Mullaitivu and Kilinochchi districts of the Northern Province, Sri Lanka(Wolters Kluwer, 2022) Wijerathna, T.; Gunathilaka, N.; Semege, S.; Pathirana, N.; Rodrigo, W.; Fernando, D.Objective: To compare the DNA sequences of Leishmania (L.) donovani isolated from individuals in two districts of the Northern Province with other parts of Sri Lanka and neighboring countries. Methods: Samples were collected from military personnel at the Army Hospital, Narahenpita, Sri Lanka from November 2018 to March 2020. A portion of the samples was fixed, stained with Giemsa and observed under the light microscope. The genomic The DNA was extracted from the remaining portion of the samples using DNEasy blood tissue kit (Qiagen, Germany) and amplified using Leishmania genus-specific primers for molecular diagnosis initially. DNA was amplified using L. donovani species-specific primers by PCR and the amplified product was sequenced for comparison of nucleotide sequences. Results: Out of 76 suspected patients, at least one biological sample of 45 (59.2%) was positive for L. amastigotes upon microscopy. Overall, 33 (43.4%) were positive in Leishmania genus-specific PCR, but only 23 (30.3%) were positive in L. donovani specific PCR. The dendrogram indicates that the current sequences clustered together with those from Nepal and Gampaha districts (Western Province), Sri Lanka, while the Indian and Eastern African sequences clustered separately. Conclusions: The genetic diversity was low among the isolates, indicating a single and possibly a local point of origin. However, the similarity of Sri Lankan and Nepal strains indicate a possibility of a shared point of origin, which needs more extensive evidence to confirm.Item The epidemiological trend of cutaneous leishmaniasis in Kegalle district, Sri Lanka: A newly established disease focus and assessment of bioclimatic suitability for disease establishment using ecological niche modelling(Elsevier, 2023) Wijerathna, T.; Wickramasinghe, K.; Gunathilaka, N.; Perera, A.; Bandara, S.Cutaneous leishmaniasis is prevalent in Sri Lanka since 1992. It remains as a health issue with increasing numbers and spreading from endemic to non-endemic areas in the county. Kegalle district is a new disease focus, which notified the first case in 2016. However, there is no documented evidence of the vector distribution, abundance and potential risk factors in this district. Two Medical Officer of Health (MOH) areas (Rambukkana and Warakapola) that had reported the highest numbers of cases in the Kagalle district was selected and recorded cases (2016-2020) were obtained. The patients were visited and socio-economic, demographic, environmental and awareness-related information was collected using an interviewer-administered questionnaire. Entomological surveys were conducted from July 2019- July 2020 using three standard entomological techniques. Bioclimatic suitability for the disease transmission was evaluated using ecological niche modelling. A total of 107 patients were reported from both MOH areas and 88 were recruited for the study. The study showed that leishmaniasis in the Kegalle district progressed to an outbreak level within 3 years after reporting the first case. School students (n = 22, 25%, P < 0.05) and individuals between 11 and 20 years of age (n = 33, 37.5%, P<0.05) were identified as the main risk groups, while the presence of composting sites (n = 65, 73.9%, P < 0.05), abandoned lands (n = 63, 71.6%, P < 0.05) in the surrounding and suboptimal (dark or normal) lighting conditions (n = 87, 98.8%, P < 0.05) in the house were identified as main risk factors. Areas closer to the adjacent district (Kurunegala), which is a known endemic district for leishmaniasis indicated a high probability (0.3-0.5) for disease transmission. Sergentomyia zeylanica (n = 164, 56.94%), was identified as the predominant sand fly species followed by Phlebotomus argentipes (n = 121, 42.01%), Sergentomyia babu insularis (n = 2, 0.69%) and Sergentomyia punjabensis (n = 1, 0.35%). Phlebotomus argentipes was captured mostly for outdoor resting sites and S. zeylanica was recoded from both indoor and outdoor. The presence of vectors that can transmit leishmaniasis in these areas along with the potential risk factors could increase the transmission risk and disease establishment in new areas that are ecologically favourable for disease transmission. Therefore, vector control entities should undertake immediate measures to stop spreading the disease into new areas.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 Morphological identification keys for adults of sand flies (Diptera: Psychodidae) in Sri Lanka(BioMed Central, 2020) Wijerathna, T.; Gunathilaka, N.BACKGROUND: Phlebotomine sand flies are a medically important group of insects that is responsible for the transmission of leishmaniasis. Surveillance plays a major role in vector control programmes through exploring species abundance, potential entomological risk and designing appropriate control measures. In field surveillance programmes of such nature, morphological identification of vector species is of paramount importance. However, in Sri Lanka, there is no published taxonomic key available for the identification of leishmaniasis vectors. METHOD: Identification keys for both male and females of the sand flies recorded in Sri Lanka were developed using morphological features. Main identification features were compared with the original observation of specimens collected from surveys and the use of published literature. Photographic illustrations of morphological features are included with the intention of making the keys user-friendly for non-taxonomists. RESULTS: A total of 22 sand fly species (Diptera: Psychodidae) of the genera Phlebotomus and Sergentomyia reported in Sri Lanka from 1910 to 2019 are included in the present work. CONCLUSION: This simplified key, along with photographs taken from specimens would be beneficial to the health staff, entomologists and research staff who deal with leishmaniasis control programmes and vector-related studies. KEYWORDS: Identification; Morphology; Sand flies; Taxonomy; Vectors.Item Histopathology of cutaneous leishmaniasis caused by Leishmania donovani in Sri Lanka.(Hindawi Pub. Co., 2020) Wijesinghe, H.; Gunathilaka, N.; Semege, S.; Pathirana, N.; Manamperi, N.; de Silva, C.; Fernando, D.ABSTRACT: Cutaneous leishmaniasis (CL) is a neglected tropical disease that is gaining importance in Sri Lanka and internationally. The clinical presentation, pathology, and method of parasite elimination in CL vary according to the species. Leishmania donovani is the causative organism for leishmaniasis in Sri Lanka. This collaborative cross-sectional study describes the clinicopathological features of cutaneous leishmaniasis among personnel of the tri-forces serving in the North and East of the country. The histology of fifty cases of CL confirmed by at least two methods (slit skin smear, lesion aspirate, tissue impression, and histology) was reviewed. The parasitic load was assessed semiquantitatively. The histological features were correlated with the clinical presentation and organism load. The majority (89.8%; n = 44) presented with a single lesion mostly located in the upper limb (69.4%). The lesion types included papule (34.7%), nodule (32.7%), and an ulcer (30.6%). The evolution time of lesions averaged 31.55 weeks. Epidermal changes were observed in 49 of the biopsies and included hyperkeratosis (90.0%; n = 45), acanthosis (44.0%; n = 22), atrophy (34.0%; n = 17), and interface change (66%; n = 33). Dermal changes were seen in all cases and were characterized by a lymphohistioplasmacytic inflammatory infiltrate of variable intensity with ill-formed granuloma in 19 cases (38%) and well-formed epithelioid granulomas in 22 cases (44%). Focal necrosis was present in 20% (n = 10). Leishmania amastigote forms were observed in 88% (n = 44). Transepidermal elimination (P = 0.025), granuloma (P = 0.027) formation, and type of lesion (P = 0.034) were significantly associated with the organism load. Granuloma formation was associated with a reduction in organism load, indicating that the macrophage activation played an important role in the control of the organism.Item A Comprehensive analysis on abundance, distribution, and bionomics of potential malaria vectors in Mannar District of Sri Lanka(Hindawi Publishing Corporation, 2019) Gunathilaka, N.; Hapugoda, M.; Wickremasinghe, R.; Abeyewickreme, W.BACKGROUND:A detailed knowledge of the distribution of the malaria vectors in Mannar district of Sri Lanka has not been studied after 1927. Past records indicated the presence of only seven species of anophelines, namely, An. culicifacies, An. subpictus, An. barbirostris, An. peditaeniatus, An. nigerrimus, An. Jamesii, and An. maculatus. There have been many changes in terms of distribution of Anopheles in the district over time. METHODS: Entomological surveillance was conducted on a monthly basis, comprising indoor hand collection, window trap collection, cattle-baited net collection, cattle-baited hut collection, and larval survey from June 2010 to June 2012 in 12 study areas under three entomological sentinel sites. The relationship between seven abiotic variables of the breeding habitats was measured. Pearson's correlation coefficients were used to determine the associations between climatic variables and anopheline densities. RESULTS:A total of 74,181 mosquitoes belonging to 14 Anopheles species were recorded. An. subpictus was the most predominant species from all techniques representing 92% (n=68,268) of the total anopheline collection. However, Anopheles culicifacies was not recorded from any site during the study period. Larval surveys identified 12 breeding habitat categories including waste water collections, lagoon water collections, and drains which were not recorded as breeding habitats by previous studies. The mean dissolved oxygen level of waste water collections was 3.45±0.15 mg/l. The mean salinity and conductivity of lagoon water collections were 21105±1344 mg/l and 34734±1974 μs/cm, respectively.CONCLUSION: The present study provides the updated knowledge on anopheline distribution and vector bionomics. Therefore, documentation of the current knowledge would be useful for learners and health authorities to design appropriate vector control measures in the prevention of reintroduction of malaria.