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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 The impact of the National Programme to Eliminate Lymphatic Filariasis on filariasis morbidity in Sri Lanka: Comparison of current status with retrospective data following the elimination of lymphatic filariasis as a public health problem(Public Library of Science, 2024) Gunaratna, I.E.; Chandrasena, T.G.A.N.; Vallipuranathan, M.; Premaratna, R.; Ediriweera, D.; De Silva, N.R.INTRODUCTION Sri Lanka implemented the National Programme for Elimination of Lymphatic Filariasis (NPELF) in its endemic regions in 2002. Five annual rounds of mass drug administration using the two-drug combination diethylcarbamazine (DEC) and albendazole led to sustained reductions in infection rates below threshold levels. In 2016, WHO validated that Sri Lanka eliminated lymphatic filariasis as a public health problem.OBJECTIVE To explore the impact of the NPELF on lymphatic filariasis morbidity in Sri Lanka.METHODS Passive Case Detection (PCD) data maintained in filaria clinic registries from 2006-2022 for lymphoedema and hospital admission data for managing hydroceles/spermatoceles from 2007-2022 were analyzed. The morbidity status in 2022 and trends in overall and district-wise PCD rates were assessed. Poisson log-linear models were used to assess the trends in PCD for endemic regions, including district-wise trends and hospital admissions for the management of hydroceles/spermatoceles.RESULTS In 2022, there were 566 new lymphoedema case visits. The mean (SD) age was 53.9 (16.0) years. The staging was done for 94% of cases, of which 79% were in the early stages (57.3% and 21.4% in stages two and one, respectively). Western Province had the highest caseload (52%), followed by the Southern (32%) and Northwestern (16%) Provinces, respectively. The reported lymphoedema PCD rate in 2022 was 0.61 per 10,000 endemic population. The overall PCD rate showed a decline of 7.6% (95%CI: 4.9% - 10.3%) per year (P < 0.0001) from 2007 to 2022. A steady decline was observed in Colombo, Gampaha and Kurunegala districts, while Kalutara remained static and other districts showed a decline in recent years. Further, admissions for inpatient management of hydroceles/spermatoceles showed a declining trend after 2015.CONCLUSIONS The PCD rates of lymphoedema and hydroceles/spermatoceles showed a declining trend in Sri Lanka after the implementation of the NPELF.Item Diversity of Wolbachia infections in Sri Lankan mosquitoes with a new record of Wolbachia supergroup B infecting Aedes aegypti vector populations(Nature Publishing Group, 2024) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S; Abeyewickreme, W.; Chandrasena, T.G.A.N.; Thayanukul, P.; Kittayapong, P.Wolbachia bacteria are common endosymbionts of insects and have recently been applied for controlling arboviral vectors, especially Aedes aegypti mosquito populations. However, several medically important mosquito species in Sri Lanka were present with limited information for the Wolbachia infection status. Therefore, the screening of Wolbachia in indigenous mosquitoes is required prior to a successful application of Wolbachia-based vector control strategy. In this study, screening of 78 mosquito species collected from various parts of the country revealed that 13 species were positive for Wolbachia infection, giving ~ 17% infection frequency of Wolbachia among the Sri Lankan mosquitoes. Twelve Wolbachia-positive mosquito species were selected for downstream Wolbachia strain genotyping using Multi Locus Sequencing Type (MLST), wsp gene, and 16S rRNA gene-based approaches. Results showed that these Wolbachia strains clustered together with the present Wolbachia phylogeny of world mosquito populations with some variations. Almost 90% of the mosquito populations were infected with supergroup B while the remaining were infected with supergroup A. A new record of Wolbachia supergroup B infection in Ae. aegypti, the main vectors of dengue, was highlighted. This finding was further confirmed by real-time qPCR, revealing Wolbachia density variations between Ae. aegypti and Ae. albopictus (p = 0.001), and between males and females (p < 0.05). The evidence of natural Wolbachia infections in Ae. aegypti populations in Sri Lanka is an extremely rare incident that has the potential to be used for arboviral vector control.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 A preliminary survey for filarial parasites among dogs and cats in mahawewa, puttalam and their vector identification(Elsevier, 2022) Nimalrathna, S.; Mallwarachchi, C.; Chandrasena, T.G.A.N.; de Silva, N.; Kimber, M.; de Silva, N.R.; Harischandra, H.PURPOSE This study investigated the prevalence of zoonotic filarial parasites within the canine and feline population in Mahawewa, Puttalam, and their vectors based on a brugian filariasis positive human case reported to the Anti-filariasis Campaign in January 2021.METHODS & MATERIALS All reachable dogs and cats, both stray and domestic, within a 500m radius of the index human case of brugian filariasis were screened for microfilaria using Giemsa stained thick blood smears prepared from capillary blood, obtained from an ear-lobe prick. Mosquito collection was done using a dog-baited trap, two window traps and a B.G. Sentinel trap from the same study site and identified using morphological keys. The head and the thorax regions of randomly selected mosquito specimens were dissected for morphological identification of larval filaria parasites via microscopy. RESULTS A total of nine dogs and three cats were surveyed, of which seven dogs and one cat had filarial infections. All the infected animals harbored B.malayi microfilariae, while four dogs and one cat were co-infected with Dirofilaria repens and two dogs with an unidentified species. A total of 119 mosquitoes were caught and identified by taxonomic keys using a dog-baited trap, two window traps and a B.G. Sentinel trap from the study site. Dissection of heads and thoraces of randomly selected 12 Mansonia annulifera, 18 Mansonia indiana 20 Mansonia uniformis and 8 Culex spp. revealed filarial larvae in M.annulifera (n=4, 33.33%), M. indiana (n=14, 77.78%), M. uniformis (n=10, 50.00%) and Culex spp.(n=5, 62.5%) via microscopy. CONCLUSION M.indiana was incriminated as a potential vector of filarial parasites for the first time in Sri Lanka. Preliminary evidence generated indicate a high prevalence of B.malayi and D.repens among dogs and cats in Mahawewa, Puttalam with an abundance of mosquito vectors mostly of Mansonia and Culex spp. This warrants further studies with a larger sample size and molecular identification of the filarial larvae within mosquito and animal samples, especially as re-emergence of brugian filariasis in humans is being reported after four decades of quiescence, and a zoonotic brugian parasite has been detected over the recent past, raising a concern from a one health perspective.Item Molecular characterization of a reemergent Brugia malayi parasite in Sri Lanka, suggestive of a novel strain(Hindawi Pub. Co., 2021) Mallawarachchi, C.H.; Chandrasena, T.G.A.N.; Withanage, G.P.; Premaratna, R.; Mallawarachchi, S.M.N.S.M.; Gunawardene, Y.I.N.S.; Dasanayake, R.S.; Gunarathna, D.; de Silva, N.R.ABSTRACT: Sri Lanka achieved elimination status for lymphatic filariasis in 2016; still, the disease remains a potential public health issue. The present study is aimed at identifying a subperiodic Brugia sp. parasite which has reemerged in Sri Lanka after four decades via molecular-based analysis. Polymerase chain reaction performed with pan-filarial primers specific for the internal transcribed spacer region-2 (ITS-2) of the rDNA of Brugia filarial parasites isolated from human, canine, and feline blood samples yielded a 615 bp band establishing the species identity as Brugia malayi. Comparison of the ITS2 sequences of the reemerged B. malayi isolates with GenBank sequences revealed a higher sequence homology with B. pahangi than B. malayi with similar phylogenetic evidence. However, the mean interspecies Kimura-2-parameter pairwise divergence between the generated Brugia sequences with B. malayi and B. pahangi was less than 3%. During the analysis of parsimony sites of the new ITS2 sequences, substitutions at A36T, A296G, T373A, and G482A made the sequences different from both B. pahangi and B. malayi suggesting the possibility of a new genetic variant or a hybrid strain of B. malayi and B. pahangi. Mosquito dissections and xenomonitoring identified M. uniformis and M. annulifera as vectors of this novel strain of B. malayi circulating among cats, dogs, and humans in Sri Lanka.Item The Diversity of Human Dirofilariasis in Western Sri Lanka(Hindawi Pub. Co, 2019) Chandrasena, T.G.A.N.; Premaratna, R.; Mallawaarachchi, C.H.; Gunawardena, N.K.; Gunathilaka, P.A.D.H.N.; Abeyewickrama, W.Y.; de Silva, N.R.BACKGROUND:Human dirofilariasis is an emerging zoonosis in many countries. Dirofilariasis caused by Dirofilaria repens may present with diverse clinical manifestations in humans due to aberrant localization of worm lesions causing diagnostic dilemmas. The aim of this retrospective study was to describe and update the demography and clinical spectrum of human dirofilariasis in western Sri Lanka. Nematode or nematode fragments isolated from excision biopsies that were confirmed as D. repens at the Department of Parasitology, Faculty of Medicine, University of Kelaniya, Sri Lanka, between 2012 and 2018 were included. Data on age, gender, and clinical details were obtained from case files. Identity of worms was established by morphometry and cuticle characteristics on wet-mount preparation. Specimens from unusual case presentations were further analyzed by PCR with specific primers for internal transcribed spacer region 2 (ITS2) of the ribosomal DNA. RESULTS:Sixteen nematode specimens isolated from subconjunctiva (n=2), subcutaneous (n=13) and intramuscular (n=1) locations were identified as D. repens by morphometry (average length 11.5 cm) and the characteristic longitudinal striations on cuticle visualized by microscopy. The age distribution of cases ranged from 1 to 65 years with a mean of 21.5. Females were more frequently affected (n=10, 62.5%) and worm locations were commonest in the orbital region (5/16) and scrotum (3/16). Imaging techniques were of use in detecting infections in deeper tissue levels. PCR analysis of DNA extracted from a worm in an intramuscular granuloma of the temporal region elicited the expected band at 484bp for D. repens. CONCLUSIONS: Human dirofilariasis is on an upward trend in incidence. Imaging techniques were of use in clinical diagnosis and molecular speciation in establishing the species identity in unusual case presentations. We suggest a more conservative approach in the management of human dirofilariasis and recommend a one health approach for control.Item Evaluation of the effects of Aedes vector indices and climatic factors on dengueincidence in Gampaha District, Sri Lanka(Hindawi Publishing Corporation, 2019) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Chandrasena, T.G.A.N.; Dassanayake, R.S.; Udayanga, N.W.B.A.L.; Abeyewickreme, W.Constant monitoring of Aedes vector indices such as Aedes mosquito abundance and ovitrap data is important for the control of dengue epidemics. Therefore, the current study attempted to evaluate the effect of larval and climatic factors on the incidence of dengue outbreaks in the Gampaha district. Based on the distribution of previously reported dengue cases, 34 households in Narangodapaluwa PHI area, Ragama, Sri Lanka, were selected randomly, and entomological surveillance was done fortnightly using adult mosquito catches and larval surveillance techniques for a period of two years. Further, weekly ovitrap surveillance was conducted for one year, by maintaining four ovitraps in a single house, two indoors and two outdoors at ground and at a height of 1.5-2 m. Based on the findings, larval indices, namely, Breteau index (BI), House index (HI), and Container index (CI), were calculated, along with the Ovitrap index (OI). The study area was positive for Ae. albopictus with an adult capturing range of 1~15/34 households. BI initially remained < 3%, which subsequently decreased up to 0. No significant difference in OI was found between the ovitraps placed at ground level and at a height of 1.5-2m (p>0.05), 95% level of confidence. The OI varied from 56.9% to 94.7% during the study period of 12 months, indicating two peaks at the monsoons. Statistics of one-way ANOVA revealed a significant difference in the monthly OI during the study period (p≤0.001) with two peaks representing the monsoonal rainfall patterns. Pearson's correlation analysis revealed that the association between dengue cases and larval indices (BI, CI, HI, and OI) and meteorological parameters was not significant (p<0.05). Migration of mosquitoes and patients could be considered as possible factors affecting the absence of a significant relationship.Item A preliminary survey of filarial parasites in dogs and cats in Sri Lanka(Public Library of Science, 2018) Mallawarachchi, C.H.; Chandrasena, T.G.A.N.; Wickramasinghe, S.; Premaratna, R.; Gunawardene, Y.I.N.S.; Mallawarachchi, N.S.M.S.M.; de Silva, N.R.Human brugian filariasis has re-emerged in Sri Lanka after a quiescent period of four decades. This study investigated the prevalence of canine and feline filarial parasites in three localities with human sub-periodic brugian filariasis, in order to determine their potential reservoir status. All reachable dogs and cats, both stray and domestic, within a 350m radius of an index case of brugian filariasis in three locations (Madampe, Wattala and Weliweriya) were screened for microfilariae using Giemsa stained thick blood smears. A representative sample of canine and feline blood samples positive for Brugia spp. microfilariae by microscopy, from each of the three locations, were further analyzed by PCR with specific primers for internal transcribed spacer region 2 (ITS2) of the ribosomal DNA. A total of 250 dogs and 134 cats were screened. The overall microfilaraemia rates were high among both dogs (68.8%) and cats (47.8%). The prevalence of microfilaraemia was significantly higher among dogs than cats (p<0.05). Two filarial species were identified based on morphology of microfilariae: Dirofilaria (Nochtiella) repens (dogs, 54.4% and cats, 34.3%) and Brugia spp. (dogs, 51.6% and cats, 30.6%). PCR analysis of canine (n = 53) and feline (n = 24) samples elicited bands in the region of 615bp, which confirmed Brugia malayi infection. Co-infection with D.(N.) repens was detected by PCR with an additional band at 484bp, in 36 canine and 17 feline samples. Overall microfilaraemia rates of dogs (81.8%) and cats (75%) in Madampe (rural) were significantly higher than in urbanized Wattala (dogs, 62.4% and cats, 26.0%) (p<0.05). High rates of zoonotic filarial infections strongly implicate dogs and cats as potential reservoirs for human dirofilariasis and brugian filariasis in Sri Lanka.Item Human infection with sub-periodic Brugia spp. in Gampaha District, Sri Lanka: a threat to filariasis elimination status?(BioMed Central, 2018) Mallawarachchi, C.H.; Chandrasena, T.G.A.N.; Premaratna, R.; Mallawarachchi, S.M.N.S.M.; de Silva, N.R.BACKGROUND: Post-mass drug administration (MDA) surveillance during the lymphatic filariasis (LF) elimination program in Sri Lanka, revealed the re-emergence of brugian filariasis after four decades. This study was done with the objectives of investigating the epidemiology and age-specific vulnerability to infection. Surveillance was done using night blood smears (NBS) and the Brugia rapid test (BRT), to detect microfilaria (MF) and anti-Brugia IgG4 antibodies in blood samples collected from an age-stratified population enrolled from two high-risk study areas (SA)s, Pubudugama and Wedamulla in the Gampaha District. The periodicity of the re-emergent Brugia spp. was characterized by quantitative estimation of MF in blood collected periodically over 24 h using nucleopore-membrane filtration method. RESULTS: Of 994 participants [Pubudugama 467 (47.9%) and Wedamulla 527 (53%)] screened by NBS, two and zero cases were positive for MF at Pubudugama (MF rate, 0.43) and Wedamulla (MF rate, 0), respectively, with an overall MF rate of 0.2. Of the two MF positives, one participant had a W. bancrofti while the other had a Brugia spp. infection. Of 984 valid BRT test readings [Pubudugama (n = 461) and Wedamulla (n = 523)], two and seven were positive for anti-brugia antibodies by BRT at Pubudugama (antibody rate 0.43) and Wedamulla (antibody rate 1.34), respectively, with an overall antibody rate of 0.91. Both MF positives detected from SAs and two of three other Brugia spp. MF positives detected at routine surveillance by the National Anti-Filariasis Campaign (AFC) tested negative by the BRT. Association of Brugia spp. infections with age were not evident due to the low case numbers. MF was observed in the peripheral circulation throughout the day (subperiodic) with peak counts occurring at 21 h indicating nocturnal sub-periodicity. CONCLUSIONS: There is the low-level persistence of bancroftian filariasis and re-emergence of brugian filariasis in the Gampaha District, Sri Lanka. The periodicity pattern of the re-emergent Brugia spp. suggests a zoonotic origin, which causes concern as MDA may not be an effective strategy for control. The importance of continuing surveillance is emphasized in countries that have reached LF elimination targets to sustain programmatic gains.Item Genotypic characterization of Orientia tsutsugamushi from patients in two geographical locations in Sri Lanka(BioMed Central, 2017) Premaratna, R.; Blanton, L.S.; Samaraweera, D.N.; de Silva, G.N.N.; Chandrasena, T.G.A.N.; Walker, D.H.; de Silva, H.J.BACKGROUND: To date more than 20 antigenically distinct strains of Orientia tsutsugamushi (OT) reported within the tsutsugamushi triangle that cause an undifferentiated acute febrile illness in humans. Genotypic characterization of OT in different geographic regions or within the same country, is important in order to establish effective diagnostics, clinical management and to develop effective vaccines. Genetic and antigenic characterization of OT causing human disease in OT-endemic regions is not known for Sri Lanka. METHODS: Adult patients and children who were admitted with an acute febrile illness and presumed to having acute scrub typhus based on presence of an eschar and other supporting clinical features were recruited. Eschar biopsies and buffy coat samples collected from patients who were confirmed having OT by IFA were further studied by real time PCR (Orientia 47 kD) and nested PCR (Orientia 56 kD) amplification. DNA sequences were obtained for 56 kD gene amplicons and phylogenetic comparisons were analyzed using currently available data in GenBank [Neucleotide substitution per 100 residues, 1000 Bootstrap Trials]. RESULTS: Twenty eschar biopsies (Location1,19, Location 2,1) and eight buffy coat samples (Location1,6, Location2,2) examined by real time PCR revealed Orientia amplicons in 16 samples. DNA sequences were obtained for the 56 kD gene amplicons in 12 eschars and 4 buffy coat samples. The genotypes of the Location1 samples revealed that, 7 exhibiting close homology with JP1 [distantly related to UT177 Thai (Karp related)], five had close homology with Kato strain, two had close homology with JGv and JG AF [Distantly related to Kawasaki M63383] and one had close homology with Gilliam strain. The Location 2 strain was closely related to Kuroki-Boryong L04956, the genotype which is distributed in far eastern Asia. Similar to other patients in the cohort this patient also had never travelled out of Sri Lanka. CONCLUSIONS: We observed all three main OT genotypes in Sri Lanka, and the majority fell into Thai Karp related clade. These results demonstrate great antigenic diversity of OT in the studied areas of Sri Lanka.