Distribution changes of cutaneous leishmaniasis: A postulated risk of disease transmission in Hambantota, Sri Lanka.

BACKGROUND OBJECTIVES: Leishmaniasis is caused by various species of parasite Leishmania. Approximately twenty of them are pathogenic to mammals. In Sri Lanka, cutaneous leishmaniasis (CL) is an established vector-borne disease. CL originates and spreads mainly through sandfly bite in many endemic countries. The aim of the present study was to compare the geographical distribution and demographic features of CL cases in Hambantota district, Sri Lanka in 2014 and 2016. METHODS: The patients who were presented to the Tangalle Base Hospital from June to December in 2014 and 2016 were examined and a descriptive study was carried out using a structured-questionnaire. Slit-skin smears were collected from each patient, Giemsa-stained and examined under the light microscope to identify Leishmania amastigotes. RESULTS: Out of 256 and 314 suspected CL patients, 156 and 155 were identified positive for the year 2014 and 2016, respectively. Out of 12 District Secretary Divisions (DSD) in Hambantota district, the highest number of CL cases, 85 and 86 was reported from Tangalle DSD in 2014 and 2016 respectively. Number of identified CL patients in Beliatta DSD had increased from 50 to 67 during the study period. In both years, majority of CL patients were ≥50 years old with males more infected than females. Although CL association with occupations were insignificant, housewives were the highly (23%) infected occupants in this area. INTERPRETATION CONCLUSION: Based on the present findings, geographical distribution within DSDs in Hambantota district had changed. This emphasizes the importance of CL as a health problem in Hambantota district.

Wolbachia infection is widespread in brackish and fresh water Aedes albopictus (Diptera: Culicidae) in the coastal Jaffna peninsula of northern Sri Lanka.

BACKGROUND & OBJECTIVES: Aedes albopictus and Aedes aegypti are important vectors of dengue and many other arboviral diseases in tropical and sub-tropical locations. Both vectors are tolerant of salinity in the dengue-endemic coastal Jaffna peninsula of northern Sri Lanka. Aedes albopictus pre-imaginal stages are found in field brackish water habitats of up to 14 parts per thousand (ppt, gL-1) salt in the Jaffna peninsula. Salinity-tolerance in Aedes is characterized by significant genetic and physiological changes. Infection with the wMel strain of the endosymbiont bacterium Wolbachia pipientis reduces dengue transmission in the field by Ae. aegypti, and the same approach is also being considered for Ae. albopictus. In this context, we investigated natural Wolbachia infections in brackish and fresh water field isolates of Ae. albopictus in the Jaffna district. METHODS: Aedes albopictus collected as pre-imaginal stages using conventional ovitraps in the Jaffna peninsula and adjacent islands of the Jaffna district were screened by PCR utilizing strain-transcending primers for the presence of Wolbachia. Wolbachia strains were then further identified by PCR using strain-specific primers for the Wolbachia surface protein gene wsp. The Jaffna wsp sequences were compared by phylogenetic analysis with other wsp sequences available in Genbank. RESULTS: Aedes albopictus were found to be widely infected with the wAlbA and wAlbB strains of Wolbachia in Jaffna. The partial wAlbB wsp surface protein gene sequence in Jaffna Ae. albopictus was identical to a corresponding sequence from South India but different from that in mainland Sri Lanka. INTERPRETATION & CONCLUSION: Widespread infection of salinity-tolerant Ae. albopictus with Wolbachia is a factor to be considered when developing Wolbachia-based dengue control in coastal areas like the Jaffna peninsula.

Reduced dengue incidence during the COVID-19 movement restrictions in Sri Lanka from March 2020 to April 2021.

BACKGROUND: Dengue is the major mosquito-borne disease in Sri Lanka. After its first detection in January 2020, COVID-19 has become the major health issue in Sri Lanka. The impact of public health measures, notably restrictions on movement of people to curb COVID-19 transmission, on the incidence of dengue during the period March 2020 to April 2021 was investigated. METHODS: The incidence of dengue and COVID-19, rainfall and the public movement restrictions implemented to contain COVID-19 transmission were obtained from Sri Lanka government sources. A Seasonal Autoregressive Integrated Moving Average (SARIMA) model was used to predict the monthly dengue incidence from March 2020 to April 2021 for each of the country's 25 districts based on five years of pre-pandemic data, and compared with the actual recorded incidence of dengue during this period. Ovitrap collections of Aedes larvae were performed in Jaffna city in the Jaffna district from August 2020 to April 2021 and the findings compared with similar collections made in the pre-pandemic period from March 2019 to December 2019. RESULTS: The recorded numbers of dengue cases for every month from March 2020 to April 2021 in the whole country and for all 25 districts over the same period were lower than the numbers of dengue cases predicted from data for the five years (2015-2019) immediately preceding the COVID-19 pandemic. The number of dengue cases recorded nationwide represented a 74% reduction from the predicted number of dengue cases for the March 2020 to April 2021 period. The numbers of Aedes larvae collected from ovitraps per month were reduced by 88.6% with a lower proportion of Ae. aegypti than Ae. albopictus in Jaffna city from August 2020 until April 2021 compared with March 2019 to December 2019. CONCLUSION: Public health measures that restricted movement of people, closed schools, universities and offices to contain COVID-19 transmission unexpectedly led to a significant reduction in the reported numbers of dengue cases in Sri Lanka. This contrasts with findings reported from Singapore. The differences between the two tropical islands have significant implications for the epidemiology of dengue. Reduced access to blood meals and lower vector densities, particularly of Ae. aegypti, resulting from the restrictions on movement of people, are suggested to have contributed to the lower dengue incidence in Sri Lanka.

Selective and electronic detection of COVID-19 (Coronavirus) using carbon nanotube field effect transistor-based biosensor: A proof-of-concept study.

In this work, we propose and demonstrate a carbon nanotube (CNT) field-effect transistor (FET) based biosensor for selective detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). CNT FETs were fabricated on a flexible Kapton substrate and the sensor was fabricated by immobilizing the reverse sequence of the RNA-dependent RNA polymerase gene of SARS-CoV-2 onto the CNT channel. The biosensors were tested for the synthetic positive and control target sequences. The biosensor showed a selective sensing response to the positive target sequence with a limit of detection of 10 fM. The promising results from our study suggest that the CNT FET based biosensors can be used as a diagnostic tool for the detection of SARS-CoV-2.

The clinical profile, hematological parameters and liver transaminases of dengue NS1 Ag positive patients admitted to Jaffna Teaching Hospital, Sri Lanka.

OBJECTIVE: Objective of the study is to evaluate the on-admission day symptoms and signs, clinical, hematological parameters and liver transaminases of the dengue NS1 positive patients who got admitted on different clinical phases [Febrile phase (day 1-3) and Critical phase(day 4-5)] of dengue at medical wards of Jaffna Teaching Hospital. RESULTS: Blood samples were collected from 150 suspected dengue patients from day 1 to 5 of the illness. Seventy-eight patients were positive for dengue NS1, according to the WHO proposed dengue clinical phase framework 37 patients were from febrile phase and 41 patients from critical phase. Patients who admitted on critical phase framework suffered from leukopenia and thrombocytopenia. Nine patients had the evidence of leakage with fever and the leakers had significant rise in hemoglobin, hematocrit and liver transaminase levels which are considered as severe form of the disease.

Salinity tolerant Aedes aegypti and Ae. albopictus-Infection with dengue virus and contribution to dengue transmission in a coastal peninsula.

BACKGROUND & OBJECTIVES: Aedes aegypti and Ae. albopictus are major arboviral vectors that are considered to lay eggs, and undergo preimaginal development only in fresh water collections. However, recently they have been also shown to develop in coastal brackish water habitats. The ability of the biologically variant salinity-tolerant Aedes vectors to transmit arboviral diseases is unknown. We therefore, investigated the infection of salinity-tolerant Aedes mosquitoes with dengue virus (DENV) and analysed dengue incidence and rainfall data to assess the contribution of salinity-tolerant Aedes vectors to dengue transmission in the coastal Jaffna peninsula in Sri Lanka. METHODS: Brackish and fresh water developing female Ae. aegypti and Ae. albopictus were tested for their ability to become infected with DENV through in vitro blood feeding and then transmit DENV vertically to their progeny. An immunochromatographic test for the NS1 antigen was used to detect DENV. Temporal variation in dengue incidence in relation to rainfall was analysed for the peninsula and other parts of Sri Lanka. RESULTS: Aedes aegypti and Ae. albopictus developing in brackish water, became infected with DENV through in vitro blood feeding and the infected mosquitoes were able to vertically transmit DENV to their progeny. Monsoonal rainfall was the discernible factor responsible for the seasonal increase in dengue incidence in the peninsula and elsewhere in Sri Lanka. INTERPRETATION & CONCLUSION: Fresh water Aedes vectors are main contributors to the increased dengue incidence that typically follows monsoons in the Jaffna peninsula and elsewhere in Sri Lanka. It is possible however, that brackish water-developing Aedes constitute a perennial reservoir for DENV to maintain a basal level of dengue transmission in coastal areas of the peninsula during the dry season, and this supports increased transmission when monsoonal rains expand populations of fresh water Aedes.

Morphological and molecular identification of cryptic species in the Sergentomyia bailyi (Sinton, 1931) complex in Sri Lanka.

The correct identification of sand fly vectors of leishmaniasis is important for controlling the disease. Genetic, particularly DNA sequence data, has lately become an important adjunct to the use of morphological criteria for this purpose. A recent DNA sequencing study revealed the presence of two cryptic species in the Sergentomyia bailyi species complex in India. The present study was undertaken to ascertain the presence of cryptic species in the Se. bailyi complex in Sri Lanka using morphological characteristics and DNA sequences from cytochrome c oxidase subunits. Sand flies were collected from leishmaniasis endemic and non-endemic dry zone districts of Sri Lanka. A total of 175 Se. bailyi specimens were initially screened for morphological variations and the identified samples formed two groups, tentatively termed as Se. bailyi species A and B, based on the relative length of the sensilla chaeticum and antennal flagellomere. DNA sequences from the mitochondrial cytochrome c oxidase subunit I (COI) and subunit II (COII) genes of morphologically identified Se. bailyi species A and B were subsequently analyzed. The two species showed differences in the COI and COII gene sequences and were placed in two separate clades by phylogenetic analysis. An allele specific polymerase chain reaction assay based on sequence variation in the COI gene accurately differentiated species A and B. The study therefore describes the first morphological and genetic evidence for the presence of two cryptic species within the Se. bailyi complex in Sri Lanka and a DNA-based laboratory technique for differentiating them.

Insecticide resistance and, efficacy of space spraying and larviciding in the control of dengue vectors Aedes aegypti and Aedes albopictus in Sri Lanka.

Unprecedented incidence of dengue has been recorded in Sri Lanka in recent times. Source reduction and use of insecticides in space spraying/fogging and larviciding, are the primary means of controlling the vector mosquitoes Aedes aegypti and Ae. albopictus in the island nation. A study was carried out to understand insecticide cross-resistance spectra and mechanisms of insecticide resistance of both these vectors from six administrative districts, i.e. Kandy, Kurunegala, Puttalam, Gampaha, Ratnapura and Jaffna, of Sri Lanka. Efficacy of the recommended dosages of frequently used insecticides in space spraying and larviciding in dengue vector control programmes was also tested. Insecticide bioassay results revealed that, in general, both mosquito species were highly resistant to DDT but susceptible to propoxur and malathion except Jaffna Ae. aegypti population. Moderate resistance to malathion shown by Jaffna Ae. aegypti population correlated with esterase and malathion carboxylesterase activities of the population. High levels of acetylcholinesterase (AChE) insensitivity in the absence of malathion and propoxur resistance may be due to non-synaptic forms of AChE proteins. Moderate pyrethroid resistance in the absence of high monooxygenase levels indicated the possible involvement of 'kdr' type resistance mechanism in Sri Lankan dengue vectors. Results of the space spraying experiments revealed that 100% mortality at a 10 m distance and >50% mortality at a 50 m distance can be achieved with malathion, pesguard and deltacide even in a ground with dense vegetation. Pesguard and deltacide spraying gave 100% mortality up to 50 m distance in open area and areas with little vegetation. Both species gave >50% mortalities for deltacide at a distance of 75 m in a dense vegetation area. Larval bioassays conducted in the laboratory showed that a 1 ppm temephos solution can maintain a larval mortality rate of 100% for ten months, and the mortality rate declined to 0% in the eleventh month. In the field, where 1 ppm concentration is gradually decreased with water usage, 100% mortality was observed only for the first four months, <50% mortality for the next two months, and 0% mortality was observed eight months after the application of temephos. Deltacide can be effectively used for space spraying programmes in Sri Lanka. Larval control can be successfully achieved through temephos with public participation.

Morphometric and meristic characterization of Phlebotomus argentipes species complex in northern Sri Lanka: evidence for the presence of potential leishmaniasis vectors in the country.

The transmission of cutaneous leishmaniasis (CL) is of public health concern in Sri Lanka. The parasite Leishmania donovani is reported to be the causative agent for CL in Sri Lanka. However there is no report on the vector of CL in the country. Phlebotomus argentipes sensu lato is the well known vector of L. donovani which causes visceral leishmaniasis (VL) in the nearby South India. The taxon Ph. argentipes previously reported to occur as a species complex comprising of two morphospecies namely A and B. The taxonomy of the Argentipes complex was reassessed recently and reported to have three species viz. Phlebotomus glaucus, Ph. argentipes sensu stricto and Ph. annandalei. A study was carried out in Jaffna mainland, where three CL patients have been recorded, and two associated islands in northern Sri Lanka to record the presence of the members of the Argentipes complex. Sandflies were collected using human landing and cattle baited collections. Collected samples were analyzed based on reported morphometric and meristic characteristics. The study revealed the presence of all three members of the complex in which Ph. glaucus and Ph. argentipes s.s. are reported for the first time in Sri Lanka.

Morphometric and meristic characterization of Phlebotomus argentipes species complex in northern Sri Lanka: evidence for the presence of potential leishmaniasis vectors in the country

The transmission of cutaneous leishmaniasis (CL) is of public health concern in Sri Lanka. The parasite Leishmania donovani is reported to be the causative agent for CL in Sri Lanka. However there is no report on the vector of CL in the country. Phlebotomus argentipes sensu lato is the well known vector of L. donovani which causes visceral leishmaniasis (VL) in the nearby South India. The taxon Ph. argentipes previously reported to occur as a species complex comprising of two morphospecies namely A and B. The taxonomy of the Argentipes complex was reassessed recently and reported to have three species viz. Phlebotomus glaucus, Ph. argentipes sensu stricto and Ph. annandalei. A study was carried out in Jaffna mainland, where three CL patients have been recorded, and two associated islands in northern Sri Lanka to record the presence of the members of the Argentipes complex. Sandflies were collected using human landing and cattle baited collections. Collected samples were analyzed based on reported morphometric and meristic characteristics. The study revealed the presence of all three members of the complex in which Ph. glaucus and Ph. argentipes s.s. are reported for the first time in Sri Lanka.

Studies on prevalence of anopheline species and community perception of malaria in Jaffna district, Sri Lanka.

BACKGROUND & OBJECTIVES: Over two decades of civil unrest and the conflict situation have had detrimental effects on vector control activities and management of malaria in Jaffna district which is an endemic region for malaria in Sri Lanka. With the background that only a few small-scale studies on malaria and its vectors have been reported from this district, a study was designed to explore the current status of malaria in the Jaffna district in relation to vector and community aspects. METHODS: Adults and larvae of anopheline mosquitoes were collected monthly from selected endemic localities. Species prevalence of the collected mosquitoes was studied while the collected adults of Anopheles subpictus, a potential vector in the district, was screened for sibling species composition based on morphological characteristics and exposed to common insecticides using WHO bioassay kits. Knowledge, attitude and practices (KAP) of the community were tested using a pre-tested structured questionnaire in high-risk and low-risk localities in the district. RESULTS: The anopheline mosquito species distribution in the district was--An. culicifacies (0.5%), An. subpictus (46%), An. varuna (4%), An. nigerrimus (44%) and An. pallidus (5.5%). Among the collected larvae the percent prevalence of An. culicifacies was 13% and other species follows as: An. subpictus (71%), An. varuna (4%), An. nigerrimus (10%) and An. pallidus (2%). Sibling species B, C and D of An. subpictus were present in the district with the predominance of B in both coastal and inland areas, while all members showed both indoor and outdoor resting characteristics, they were highly resistant to DDT (4%) and highly susceptible to malathion (5%). KAP study in the district showed a reasonable level of knowledge, positive attitude and practices towards malaria. CONCLUSION: An. subpictus, the reported major vector of Jaffna and a well-established secondary vector of malaria in the country, continues to be the predominant anopheline species. The distribution of sibling species of An. subpictus complex in the Jaffna district, revealed for the first time, has implications for future studies on its bionomics and malaria transmission pattern in this area and the planning of control strategies for this region. The community perception of disease, which revealed a satisfactory knowledge indicates the potential for better community participation in future malaria control activities in this region. As potential vectors are still present, health authorities need to be vigilant to prevent any future epidemics of malaria.

Perception and personal protective measures toward mosquito bites by communities in Jaffna District, northern Sri Lanka.

Mosquito-borne diseases are of public health importance in war-torn northern Sri Lanka. The severity of mosquito bites and attitudes of the public toward mosquito problems were investigated using a structured questionnaire among communities in 3 administrative divisions in Jaffna District. One hundred fifty-four households were interviewed during this study. Sixty-four percent of the respondents reported that the mosquito problem was severe in their localities. Fifty-two percent stated that mosquito-biting activity was severe in the evening (1500 h-1900 h), 41% at night (after 1900 h), and 7% throughout the day. Severity of mosquito menace was found to have no association with type of house construction. Seventy-seven percent were able to name at least 1 disease transmitted by mosquitoes. Statistical analysis showed no association between education level and public awareness on mosquito-borne diseases. Nearly 88% were able to identify at least a breeding source of mosquitoes and most of them practice measures to eliminate suitable environments for mosquito breeding. Ninety-six percent used personal protective measures against mosquito bites during some seasons or throughout the year. Mosquito coils were the most commonly used personal protective method followed by bed nets. The monthly expenditure for personal protective measures varied from US$0.19 (LKR 20) to US$3.40 (LKR 350).

Anopheles culicifacies sibling species B and E in Sri Lanka differ in longevity and in their susceptibility to malaria parasite infection and common insecticides.

Members of the Anopheles culicifacies Giles complex (Diptera: Culicidae) are well established as the predominant vectors of malaria in Sri Lanka. Until recently, only sibling species B was reported to be present in Sri Lanka, which was surprising as species B is a poor vector of malaria in India. This was clarified by the identification through Y-chromosome morphology that what was reported as B on the island is really a mixture of B and E. The fecundity, longevity and insecticide resistance of B and E are of relevance to malaria transmission and its control and are reported in this study. The mean egg production of these two sibling species did not differ significantly. The mean age of wild mosquitoes was assessed by the Polovodova technique of observing ovarian dilatations. More of species E than B had three or more dilatations, i.e. had reached an age at which sporozoites could have developed to maturity, although the difference between the species was of borderline significance. Following feeding on Plasmodium vivax or Plasmodium falciparum infected blood, some females of species E developed oocysts but none of species B did so. Both sibling species were found fully susceptible in laboratory tests to lambdacyhalothrin and deltamethrin, but resistant to DDT and partially resistant to malathion.

Some characteristics of the larval breeding sites of Anopheles culicifacies species B and E in Sri Lanka.

BACKGROUND & OBJECTIVES: Anopheles culicifacies Giles, the major malaria vector in Sri Lanka, exists as a species complex comprising two sympatric sibling species--species B and E. Species E is reported to be the major vector of Plasmodium vivax and P. falciparum parasites in Sri Lanka, whilst species B is a poor or nonvector as in India. Knowledge of the breeding habits of the two sibling species can help in designing optimal vector control strategies. Hence, a survey was conducted in Sri Lanka to study the preferential breeding habitats of An. culicifacies species B and E. METHODS: Immature forms of An. culicifacies were collected from identified breeding sites in malarious districts. Collected larvae were typed for their sibling species status based on mitotic Y-chromosome structure. Data was analysed using Statistical Package for Social Science version 10.0. RESULTS: An. culicifacies immature forms were found in 23 collection sites. Among these samples 19 were found to have species E and four to have species B. All species B larvae were collected from Tonigala village in the Puttalam district. None of the 23 sites was found to have both species B and E. Species E, the major vector of malaria, appears to breed in variety of breeding sites which can be of an indication of its adaptive variation to exploit breeding sites with varying limnological characteristics. INTERPRETATION & CONCLUSION: The present findings have to be taken into account when formulating more effective larval control measures. They also show the need for a detailed study of possible different preferences for larval breeding sites between species B and E.

Molecular and biochemical characterization of a sand fly population from Sri Lanka: evidence for insecticide resistance due to altered esterases and insensitive acetylcholinesterase.

With an increasing incidence of cutaneous leishmaniasis in Sri Lanka, particularly in northern provinces, insecticide-mediated vector control is under consideration. Optimizing such a strategy requires the characterization of sand fly populations in target areas with regard to species composition and extant resistance, among other parameters. Sand flies were collected by human bait and cattle-baited net traps on Delft Island, used as an illegal transit location by many refugees returning to the north of Sri Lanka from southern India where leishmaniasis is endemic. For species identification, genomic DNA was extracted and a fragment of the ribosomal 18S gene amplified. The sequence from all flies analysed matched that of Phlebotomus argentipes Annandale & Brunetti, the primary vector in India and the most likely vector in Sri Lanka. Independent morphological analysis also identified P. argentipes. To establish the current susceptibility status of vector species, data were obtained at the biochemical level, from which potential cross-resistance to alternative insecticides can be predicted. The Delft Island collection was assayed for the activities of four enzyme systems involved in insecticide resistance (acetylcholinesterase, non-specific carboxylesterases, glutathione-S-transferases and cytochrome p450 monooxygenases), establishing baselines against which subsequent collections can be evaluated. There was preliminary evidence for elevated esterases and altered acetylcholinesterase in this population, the first report of these resistance mechanisms in sand flies to our knowledge, which probably arose from the malathion-based spraying regimes of the Anti-Malarial Campaign.

Anopheles culicifacies Y-chromosome dimorphism indicates sibling species (B and E) with different malaria vector potential in Sri Lanka.

In Sri Lanka, malaria is transmitted mainly by Anopheles culicifacies Giles sensu lato (Diptera: Culicidae). In India, this nominal taxon comprises sibling species A, B, C, D and E, distinguished by their chromosome morphology. Species B (identified by polytene chromosome sequence Xab, 2g1 + h1) is not such an efficient vector of malaria as other members of the An. culicifacies complex in India. All specimens of An. culicifacies s.l. examined from Sri Lanka possess Xab, 2g1 + h1 polytenes, previously interpreted as species B, despite their important vector status. Recently, species E was described from Rameshwaram Island (Tamil Nadu, India) between Sri Lanka and the Indian mainland, where both species B and E are sympatric. Species B and E share polytene sequence Xab, 2g1 + h1 but differ by the mitotic Y-chromosome being acrocentric in species B, submetacentric in species E, the latter implicated as vector of vivax malaria. From May 1999 to January 2000, we surveyed Y-chromosomes of male progeny from An. culicifacies Xab, 2g1 + h1 females collected from cattle bait in diverse malarious districts of Sri Lanka: Badulla, Monaragala, Puttalam and Trincomalee. Karyotypes of readable quality were obtained from 42/83 families examined, with overall proportions 24% acrocentric and 76% submetacentric Y-chromosome carriers, both types being sympatric in at least 3/4 localities sampled. By analogy with the situation on Rameshwaram Island, we interpret these observations to demonstrate widespread presence of two members of the An. culicifacies complex in Sri Lanka, their karyotypes being compatible with species B and E, the latter predominant and having greater vector potential.