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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 Intestinal parasitoses and the nutritional status of internally displaced children in Vavuniya(Sri Lanka Medical Association, 2005) Chandrasena, T.G.A.N.; Hapuarachchi, H.A.C.; Dayanath, M.Y.D.; de Silva, N.R.OBJECTIVES: To assess the prevalence of intestinal parasitic infections and the nutritional status of children of internally displaced families residing in refugee camps. MeTHODOLOGY: Saline smears and modified Kato Katz techniques were performed on stool samples collected from children of displaced families residing in the Adappankulam refugee camp in Vavuniya. The heights and weights of the children were measured and standard anthropometric indices, weight- for-height (WH), Height- for- age (HA) and weight- for- age (WA) Z scores were calculated using Epi Info. RESULTS: Stool samples of 159(83 males) of 200 children registered at Adappankulam refugee camp were screened for intestinal parasites. The mean age of the study population was 7.0 years (range 2-15 years). One or more intestinal parasites were detected in 40.25 % (64/159). Twenty point one percent had helminth and 24.5% (39) had protozoan infection. Of 32 children with helminth infections, 29(18.2%) had hook worm, 2(1.25%) Ascaris lumbricoides, 3 (1.8%) Trichuris trichiura and 1(0.62%) Enterobius .vermicularis infections. The most common pathogens were hookworm and Giardia lamblia (23, 14.5%). The anthropometric indices of 161 children (100 males) were calculated. Of the 105(65.2%) children with growth retardation, 76(47.7%) were wasted 56(34.7%) stunted and 122(75.7%) underweight. There was no significant correlation of the mean Z scores with Giardia or hookworm infection. CONCLUSIONS: There was an elevated prevalence of growth retardation among this group of displaced children. The prevalence of'Giardia and hookworm infections was moderately high. Other pathogenic intestinal parasites were scarce in this community.Item Recent chikungunya outbreak in Sri Lanka 2006-2007(Faculty of Tropical Medicine, Mahidol University, 2007) Abeyewickreme, W.; Bandara, K.B.A.T.; Dayanath, M.Y.D.; Sumanadasa, D.; Hapuarachchi, H.A.C.; Gunawardena, N.K.; Hapugoda, M.D.; Wijesiriwardena, B.; de Silva, S.; Perera, T.BACKGROUND: Chikungunya(CHIK) is a viral disease transmitted by Aedes mosquitoes. Cases with symptoms of CHIK had been reported from several parts of Sri Lanka in 2006-2007. Laboratory testing of samples is a prime requirement for confirmation of transmission. OBJECTIVES: To confirm CHIK infection in suspected patients by rapid Reverse Transcription Polymerase Chain Reaction Assay(RT-PCR), find out manifestations specific for CHIK infection and study the transmission of CHIK virus by vector mosquitoes. METHODOLOGY: Serura. samples and information on clinical manifestations were collected from 189 chikungunya-suspected patients from different geographical areas in Sri Lanka from September 2006 to September 2007. Samples were tested for Chikungunya RNA by RT-PCR. Amplified products were visualized by agarose gel electrophoresis. Adult mosquitoes were also collected from chikungunya case-reported stations. They were tested for Chikungunya RNA through RT-PCR-followed by agarose gel electrophoresis assay. RESULTS: Of the CHIK-suspected patients reported from all parts of the island 86/189 (45.5%) were positive for CHIK virus. Of the PCR positive 06, all had fever with either arthralgia or arthritis or both. Headache (95.3%) and backache (84.6%) were also common among above patients. Eight percent (4/50) of both species of Aedes mosquitoes were RT-PCR positive. DISCUSSION: RT- PCR is important in early diagnosis of the infection and differentiation from dengue fever. The most common clinical symptoms observed were fever with either arthralgia, arthritis or both. Both Aedes aegypti and Aedes. albopictus are important in transmitting the disease.Item A Case report of dengue and chikungunya co-infection in Sri Lanka(The Parasitology and Tropical Medicine Association of Thailand, 2008) Abeyewickreme, W.; Hapuarachchi, H.A.C.; Bandara, K.B.A.T.; Hapugoda, M.D.; Williams, S.Dengue fever and chikungunya are arboviral diseases transmitted by Aedes mosquitoes. Though dengue has been an important communicable disease in Sri Lanka for many years, chikungunya has not been reported in Sri Lanka since late 1960s. However, in November 2006, an outbreak suggestive of chikungunya erupted in the country. We report here the first laboratory confirmed case of dengue and chikungunya co-infection in Sri Lanka. The objective is to confirm the co-infection of dengue and chikungunya in a clinical case reported in November 2006. Clinical history of high fever, severe headache, nausea, loss of appetite, severe arthralgia and mild oedema of knees, small joints of hands and feet for 3 days suggested the possibility of dengue and chikungunya in a 70 year old male. There was no skin rash or bleeding manifestations. Laboratory investigations performed included total white blood corpuscle count/differential count (WBC/DC), platelet count (PLT), serum, haemoglobin (Hb%) and packed cell volume levels (PCV). Reverse Transcription- Polyrnerase Chain Reaction (RT-PCR) technology was used to confirm the presence of either dengue or chikungunya. Viral RNA was extracted from serum samples collected during the first five days of infection using QiAmp Viral RNA Kits and amplified products were visualized by 2% agarose gel electrophoresis and ethidium bromide staining. WBC/DC analysis showed a leucopaenia (WBC count 3.04 x 103 per μl) with relative lymphocytosis (51.0%). The total PLT was 115 x 103 per μl. Hb% was 14.3 g/dl with a PCV of 43.8%. The presence of both infections was confirmed by RT-PCR which amplified 225 bp and 354 bp products for dengue and chikungunya respectively. This was the first laboratory confirmed case of dengue and chikungunya co-infection, which was also the first confirmed report of chikungunya since 1969 in Sri Lanka. As clinical and biochemical manifestations of this patient suggested the probability of a mixed infection of dengue and chikungunya, the confirmation was achieved by a RT-PCR assay. This report highlights the importance of using molecular assays to confirm mixed viral infections during their early stages, especially infections such as dengue which can result in fatal complications.Item A Mixed infection of Plasmodium falciparum and Plasmodium malariae: the first report of a Plasmodium malariae infection after 37 years of its absence in Sri Lanka(2008) Hapuarachchi, H.A.C.; Abeysundara, S.; Gunawardena, N.K.; Manamperi, A.; Senevirathne, M. P.; Leemingsawat, S.; Chavalitshewinkoon-petmitr, P.; de Silva, N.R.; Abeyewickreme, W.Malaria has been endemic in Sti Lanka for several centuries. Currently, only Plasmodium falciparum and P. vivax are present in the country. P. malariae infections have not .been reported in Sri Lanka since 1969. The objective is to determine the presence of malaria species in a patient returned from Malawi. The clinical history of intermittent high fever for 2 weeks accompanied by severe headache, myalgia, arthralgia, vomitimg, loss of appetite and backache with ictetus and mild hepatosplenomegaly suggested malaria in this 51 year old patient. Apart from the basic biochemical investigations, presence of malarial species was determined by light microscopy and confirmed by Real-Time Polymerase Chain Reaction (PCR) technology. Biochemical investigations showed a high serum bilirubin (4.8 mg/di) and liver enzyme (SGOT = >125 units, SGPT = >250 units) levels. Serum haemoglobin level (12.8 g%) was normal. Except for the presence of ptoteinuria (albumin = ++), bile (+) and red blood corpuscles (RBC) in his urine, renal functions were normal. Microscopical examination of Giemsa stained thin and thick blood smears showed an asexual parasite density of 120,000 per ul of blood. Infected RBCs were not enlarged, The presence of double-chromatin and applique form trophozoites, occasionally invading multiple RBCs suggested P. falciparum infection. In addition, there were characteristic band form trophozoites of P. malariae. Real-Time PCR protocol confirmed the presence of both P. falciparum and P. malariae in this patient. This is the first case of P. malariae reported in Sri Lanka after 4 decades, though the infection had been acquired from Malawi. Clinical and biochemical evidence indicated liver dysfunction and a transient glomerulonephritis, both of which subsided after treatment with quinine. This case report emphasizes the need of physicians to be more vigilant about the presence of malaria among immigrants, despite the drastic reduction of malaria in the country in recent years. Hence, this report highlights the importance of a proper programme in Sri Lanka to screen immigrants for infectious diseases.Item K76T point mutation of chloroquine resistance transporter gene: Is it a potential molecular marker for chloroquine resistance in Sri Lankan Plasmodium falciparum isolates?(Sri Lanka Association for the Advancement of Science, 2007) Hapuarachchi, H.A.C.; Dayanath, M.Y.D.; Abeysundara, S.; Manamperi, A.; Abeyewickreme, W.; de Silva, N.R.Current evidence suggests that K76T mutation of chloroquine resistance transporter (Pfcrt) gene may be used as a molecular marker for chloroquine (CQ) resistance of P. falciparum. This study was carried out to determine the frequency of K76T mutation of Pfcrt gene in Sri Lankan P. falciparum isolates collected from Mannar district in the Northern Province. Mutation patterns were compared with in vitro and in vivo CQ failure rates for this parasite population to analyze its association with resistance to CQ and to calculate the genotype-resistance (GRI) and genotype-failure (GFI) indices for K76T mutation. P. falciparum DNA was extracted from dried blood spots using a QiaAmp DNA Blood Mini Kit. Mutation patterns at 76 codon of Pfcrt of field isolates were detected using a polymerase chain reaction - restriction fragment length polymorphism assay. Parasite isolates were categorized into wild (sensitive) and mutant (resistant) types based on the banding patterns of digested products on 2% agarose gels. GRI and GFI indices were calculated for this parasite population. Of 38 CQ resistant isolates, 86.8% (N = 33) showed the mutant allele (K76T) at codon 76 of Pfcrt. There was a statistically significant association between the presence of K76T mutation and in vivo resistance to CQ (x2 = 5.11, p = 0.02). Of 33 sensitive isolates, 39.4% (N = 20) possessed the wild type allele at the same codon position. The calculated GRI and GFI indices were 1.13 and 1.38 respectively for this area. Even though results show a statistically significant association between the presence of K76T allele and CQ treatment failure of P. falciparum isolates in the study population, its mere presence alone does not seem to correlate with resistance to CQ. Therefore, mutations at other codons of Pfcrt shown to accompany K76T mutation as well as those in P. falciparum multi drug resistance 1 (Pfmdr1) gene need to be analyzed to determine whether other mutations play a role together with K76T in clinically resistant Sri Lankan parasite isolates. More studies are required to validate the calculated GRI and GFI values, which may be used to predict the therapeutic failure of CQ in a particular area in Sri Lanka in the future. Acknowledgement: National Science Foundation Research grant SIDA/2005/BT/03 and by the IAEA TC Project SRL 06/028Item Potential use of blood-fed mosquitoes as evidence in forensic casework(Sri Lanka Association for the Advancement of Science, 2007) Marasinghe, M.P.L.R.; Sirisena, D.M.; Bandara, K.B.A.T.; Hapuarachchi, H.A.C.; Abeyewickreme, W.Medico - criminal entomology has been an important source of evidence in forensic investigations for many years. However, it has not been widely used to provide direct evidence for personal identification in forensic casework so far. In this study, we made an attempt to determine the usefulness of human DNA extracted from blood-fed mosquitoes as evidence in forensic casework. Approximately 1500 adult female mosquitoes of same age from four different species, i.e. Culex quinquefasciatus, Armigerus sabalbatus, Aedes aegypti and Anopheles tessellatus were fed with the same volume of human blood and maintained under the same environmental conditions. Three batches (N=1, 5 and 10) of randomly selected blood-fed mosquitoes were crushed and blotted onto filter paper strips separately at two hour intervals subsequent to blood meals up to 48 hours to determine the longitudinal variation in the extraction of polymerase chain reaction (PCR) amplifiable human DNA from mosquitoes. Human DNA was extracted from filter papers using a chelex-100 extraction protocol and amplified by PCR technique using human primers. Amplified products were run in 1.5% agarose gels. PCR amplifiable human DNA could be extracted from mosquitoes of Cx. quinquefasciatus and An. tessellatus up to 48 hours subsequent to blood meals. However, this duration was up to 46 and 42 hours in Ae. aegypti and Ar. Sabalbatus, respectively. The amount of amplifiable DNA was inversely proportionate to the post-feeding duration, but increased with number of mosquitoes at each time interval. Amplifiable human DNA could be extracted even from a single mosquito of four different species even after 48 hours from a blood meal with slight variation among species. However, as the amount of DNA decreases with time interval, more mosquitoes will have to be used for extraction in late sample collections. Similarly, it may be required to use a highly sensitive PCR protocol when analyzing such samples. Blood-fed mosquitoes collected even after 2 days from a crime scene may be used as a source of direct evidence for personal identification in forensic casework.Item Night blood survey of a selected high-risk population for lymphatic filariasis(Sri Lanka Association for the Advancement of Science, 2007) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Abeyewickreme, W.; Gunawardena, N.K.; Hapuarachchi, H.A.C.; Abeysundara, S.Human infection with Wuchereria bancrofti causes a disabling parasitic disease known as lymphatic filariasis, which is a major public health and socio-economic problem in many parts of the world. Little is known about the prevalence of filariasis among high-risk populations for filariasis. Objective of this study was to determine such prevalence of lymphatic filariasis among Mahara prison inmates whom the Anti Filaria Campaign (AFC) has identified as a high-risk group. All inmates of Mahara Prison were screened for Microfilariae (Mf) except those in special cells, by night blood film microscopy to determine the prevalence of infection from February to May 2007. All inmates were males of greater than 15 years. Of the 423 inmates screened, 15 were positive for Mf, giving a Mf positive rate of 3.55% in the study population and a mean Mf density of 5 Mf/60 æl blood, ranging between 4 to 9.2 Mf /60 æl of blood with a standard deviation of 2.49. The highest number of infected inmates was residents of Colombo and Gampaha districts where transmission is currently taking place. This is one of the few studies undertaken to date to determine the prevalence of bancroftian filariasis among inmates of a prison, a neglected population in Sri Lanka. This study indicates that the Mf rate of bancroftian filariasis in this study population is far greater than the 0.18% currently reported in the country. Therefore, an intensive programme is recommended to contain the spread of infection within this study population. For this, a proper screening programme combined with antifilarial treatment and vector control programme is urgently required. Acknowledgements: Authors wish to acknowledge the financial assistance received from WHO/SEARO/TDR (grant no. SN 1152) and University of Kelaniya (Research grant no. RP/03/04/06/01/2006). Authors wish to thank Dr. Ravi Mudaliage, Senior Medical Officer, Prison's Hospital, Mahara, Ragama for his support and encouragement during field study activities. Authors also wish to thank Mr. M. Y. D. Dayanath, Ms. N.M. Ashoka Malanie, Mr. M.I.M.Peris, Mr. Y.L.Rassapana and other staff members of the Molecular Medicine Unit and Department of Parasitology, Faculty of Medicne, University of Kelaniya, Ragama for their assistanceItem Molecular markers of chloroquine resistance in Plasmodium falciparum in Sri Lanka:frequency before revision of the antimalarial drug policy(Academic Press, 2009) Hapuarachchi, H.A.C.; Abeysundara, S.; Dayanath, M.Y.D.; Manamperi, A.; Abeyewickreme, W.; de Silva, N.R.No Abstract AvailableItem Laboratory confirmation of dengue and chikungunya co-infection(Sri Lanka Medical Association, 2008) Hapuarachchi, H.A.C.; Bandara, K.B.A.T.; Hapugoda, M.D.; Williams, S.; Abeyewickreme, W.No Abstract AvailableItem A Case of imported malaria: the first report of Plasmodium malariae infection in Sri Lanka after 37 years(Sri Lanka Medical Association, 2008) Hapuarachchi, H.A.C.; Gunawardena, N.K.; Senevirathne, M.P.; Abeyewickreme, W.; de Silva, N.R.We report a case of Plasmodium falciparum and P. malariae mixed infection in a patient who had been living in Malawi. This is the first case of P. malariae reported in Sri Lanka in 4 decades. The presence of both parasites was confirmed by microscopy and polymerase chain reaction (PCR). The history strongly indicated that the infection had been acquired from Malawi. The patient had liver dysfunction and a transient glomerulonephritis, both of which subsided with antimalarial treatment.