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Naturally Occurring Microbiota Associated with Mosquito Breeding Habitats and Potential Parasitic Species against Mosquito Larvae: A Study from Gampaha District, Sri Lanka.

ABSTRACT: A mosquito species has its own favourable requirements of abiotic and biotic characteristics including microbiota, in a breeding habitat. Some of the microbiota may cause parasitic or pathogenic effects to mosquito larvae such as species of viruses, parasitic bacteria, fungi, protists, entomopathogenic nematodes, and filamentous fungi. In Sri Lanka, there is a scarcity of information on microbiota associated with mosquito breeding habitats and their effect on mosquito larvae. Hence, the present study was conducted to determine microbiota species/taxa associated with a variety of mosquito breeding habitats in selected areas of the Gampaha District in Sri Lanka and the relationship, if any, the microbiota has with mosquito larva survival and breeding. Forty-five microbiota species belonging to 11 phyla were found from different mosquito breeding habitats with the highest percentage belonging to phylum Euglenozoa (27.89%). Species that belonged to the phylum Amoebozoa (1.22%) and Sarcodina (1.17%) had the lowest abundance, and each of its species richness was recorded as one. Philodina citrina followed by Monostyla bulla comprised 30.8% and 16.59%, respectively, of the total rotifer population. From the total microbiota, 25-50% existed as accidental while less than 25% rare, in the habitat type according to their abundance. Paddy fields had the highest species richness (17), evenness (23.52), Shannon-Weiner (66.64), and beta diversity (0.65) over 50% indicating high heterogeneity in microbiota composition among the habitats. Ciliated protists, namely, Vorticella microstoma, Zoothamnium spp., and Chilodinella sp., were identified as naturally occurring microbiota associated with Culex mosquito larvae that inhabited in paddy fields and associated irrigation canals. Only Vorticella microstoma caused a significant lethal effect on mosquito larvae. This study revealed that species of Cx. gelidus, Cx. pseudovishnui, Cx. tritaeniorhynchus, Cx. quinquefasciatus, and Cx. whitmorei served as hosts for V. microstoma where infectivity rate in Cx. tritaeniorhynchus reached 73.22. Chilodinella sp. selectively served as endoparasitic to Cx. gelidus larvae causing only 4.58% mortality, and invasive cysts of the pathogen were observed in the subcuticular layer of the host body. Even though Zoothamnium spp. were found on Cx. tritaeniorhynchus larvae, there was no lethal effect due to the attachment of the parasitic agent. The potential of these microbiotas in integrated vector controlling approaches in future perspectives is recommended.

SUBMITTER: Ranasinghe HAK

PROVIDER: S-EPMC7204370 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Naturally Occurring Microbiota Associated with Mosquito Breeding Habitats and Potential Parasitic Species against Mosquito Larvae: A Study from Gampaha District, Sri Lanka.

Ranasinghe H A K HAK Amarasinghe L D LD

BioMed research international 20200428

A mosquito species has its own favourable requirements of abiotic and biotic characteristics including microbiota, in a breeding habitat. Some of the microbiota may cause parasitic or pathogenic effects to mosquito larvae such as species of viruses, parasitic bacteria, fungi, protists, entomopathogenic nematodes, and filamentous fungi. In Sri Lanka, there is a scarcity of information on microbiota associated with mosquito breeding habitats and their effect on mosquito larvae. Hence, the present ...[more]

PMID: 32420344

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Project description:BackgroundThe 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.MethodsAdults 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.ResultsAdults 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.ConclusionsThis 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).

Project description:BackgroundOrthohantaviruses and leptospira are emerging zoonotic pathogens of high public health significance. The epidemiology of orthohantavirus infections and leptospirosis is similar and presents related clinical pictures in humans. However, a paucity of data on actual reservoir hosts for orthohantaviruses and leptospira exists. Therefore, this study aimed at determining the occurrence of orthohantaviruses and leptospira in small mammals captured in an endemic region of Sri Lanka.MethodsRodents and shrews were morphologically and/or genetically identified using morphological keys and DNA barcoding techniques targeting the cytochrome oxidase b subunit gene (Cytb). Lung tissues and sera were subsequently analyzed for the presence of orthohantavirus RNA using qRT-PCR. Sera of rats were tested for IgG antibodies against orthohantaviruses and leptospira.ResultsForty-three (43) small mammals representing: Rattus (R.) rattus (black rat) or R. tanezumi (Asian rat), Suncus murinus (Asian house shrew), R. norvegicus (brown rat) and Mus musculus (house mouse) were investigated. No orthohantavirus RNA was detected from the lung tissue or serum samples of these animals. Elevated levels of IgG antibodies against Puumala orthohantavirus (PUUV) and/or Seoul orthohantavirus (SEOV) antigens were detected in sera of 28 (72%) out of the 39 rats analysed. Interestingly, 36 (92%) of the 39 rats also showed presence of anti leptospira-IgG antibodies in their serum, representing dual infection or dual exposure in 26/39 (66.7%) of examined rats.ConclusionsThis project targets important public health questions concerning the occupational risk of orthohantavirus infections and/or leptospirosis in an endemic region of Sri Lanka. Most rats (72%) in our study displayed antibodies reacting to orthohantavirus NP antigens, related to PUUV and/or SEOV. No correlation between the orthohantavirus and leptospira IgG antibody levels were noticed. Finally, a combination of both morphological and DNA barcoding approaches revealed that several species of rats may play a role in the maintenance and transmission of orthohantavirus and leptospira in Sri Lanka.

Project description:Dengue is the most important mosquito-borne viral infection in Sri Lanka causing an enormous social and economic burden in the country. In the absence of therapeutic drugs and the developed vaccines are under investigation, vector control is the best strategy to reduce the disease transmission. Therefore, the development of novel tools to control dengue vector mosquitoes has become the need of the hour. Novaluron is a recently developed Insect Growth Regulator (IGR) which inhibits chitin synthesis in immature stages of insects. The aim of the study was to identify the efficacy of a simple and cost-effective Autocidal Gravid Ovitrap (AGO) developed using Novaluron to control dengue outbreaks in the District of Gampaha, Sri Lanka. Laboratory and semifield experiments were performed to identify the activity range, optimum field dosage, and residual effects of Novaluron following the World Health Organization guidelines, and field experiments were performed in the Ragama Medical Officer of Health (MOH) area. Two study areas 800?m apart were selected and assigned as treated and control areas randomly. In each study area, 30 households were selected randomly. Each household was given two ovitraps, one placed indoors and the other placed outdoors. Mortality and survival counts were recorded separately for one-year time period and data were analyzed using a two-way repeated measures analysis of variance model. During the laboratory experiments, the adult emerging inhibition was 100% in all tested concentrations. The optimum field dosage was 2?ppm and the residual effect was 28 days. In the field experiments, significantly higher mortality counts were recorded in treated areas both indoor- and outdoor-placed AGOs. Two-factor repeated measures ANOVA followed by Tukey's test confirmed that the mean mortality count is high for the developed AGOs both indoor and outdoor settings. The developed AGO can be deployed to control both indoor and outdoor dengue vector mosquito populations, and in dengue-risk areas, the ovitrap will be supportive to local health authorities to enhance the efficiency of future vector control programs.

Project description:BackgroundLeishmaniasis is a notifiable disease in Sri Lanka since 2008. Previous studies show a gap in the notification of leishmaniasis. The purpose of the present study was to determine the Knowledge, attitudes and practice of medical officers regarding leishmaniasis.MethodsA cross-sectional study was conducted in the Anuradhapura district which reported the highest case load of leishmaniasis. Medical officers from public and private health care institutes in the area filled a self-administered questionnaire in the presence of the investigators.ResultsOne hundred and eighty-eight (188) medical officers completed the questionnaire. Of them, 95.7% were aware of leishmaniasis as a parasitic infection and 84.7% correctly identified Leishmania donovani as the causative organism in Sri Lanka. From the respondents, 181 (96.8%) knew that the vector of leishmaniasis is sand fly. Cutaneous leishmaniasis was reported as the most prevalent form of leishmaniasis in the country by 176 (94.1%). Nearly half of the respondents (98, 54.1%) were aware of the fact that the Anuradhapura district has the highest disease burden. Many of them had the idea that leishmaniasis is an emerging disease (155, 84.3%,) and early diagnosis is important in controlling the disease (163, 89.1%). Although about three fourth (123, 73.7%,) of the participants mentioned that leishmaniasis should be notified at first clinical suspicion, only 74 (42.5%) were aware that it is a legal requirement. Some medical officers (39, 22%) believed that the current notification system in the country is not effective. Unavailability of notification forms (60, 36.8%) heavy workload (85, 50.3%) and inadequate supportive staff (55, 35.1%) were reported as barriers for timely notification. Even though 105 (58.0%) of medical officers had suspected leishmaniasis during the last 8 years period only 35 (19.4%) had notified.ConclusionsEven though more than 90% of the participants had good theoretical knowledge about leishmaniasis; notification of leishmaniasis is considerably inadequate. This study emphasizes the need for greater efforts to improve the notification of leishmaniasis in Sri Lanka.

Project description: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, ? 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 Bacillaceae family (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.

Project description:BackgroundThe 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.MethodsLarval 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.ResultsTotal 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.ConclusionsThe 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.

Project description:BACKGROUND:Sand flies are responsible for the transmission of several disease pathogens including Leishmania. Sand flies breed in habitats with high levels of humidity and organic matter. They are nocturnal in nature and peak activity ranges from dusk to dawn. The scientific evidence on breeding ecology and diurnal resting sites of sand fly fauna are important aspects of planning and implementing vector control activities. However, such fundamental information is grossly inadequate in Sri Lanka to support the control efforts in the country. Therefore, the present study addresses some of the important aspects of sand fly breeding ecology and diurnal resting sites. METHODS:Potential resting sites were thoroughly observed, and sand flies were collected using a battery-operated aspirator and sticky papers when appropriate from three selected Medical Officer of Health (MOH) areas (Polpithigama, Maho and Galgamuwa) in Kurunegala district, Sri Lanka. Soil samples were collected from each potential breeding site. Half of each soil sample was incubated for 45 days. The other half was screened for immature stages. Adult sand flies collected from field and emerged adults at the insectary under confined incubation were identified using morphological characteristics. RESULTS:Pepper bushes and termite mounds were the most notable resting sites while, betel bushes, cattle huts, piles of coconut shells, latrines, manna bushes and tree holes were also positive for sand fly adults. Only two species, Phlebotomus argentipes and Sergentomyia punjabensis, were reported. Soil samples were collected from a total of 432 sites and 7 of them were positive for immature stages. Predominant breeding habitats identified during the present study were mud flats and moist soils of rice paddies, the soil below decaying hay, drying irrigational tank bottom moist soil, and the floors of cattle huts. CONCLUSION:This study demonstrates that the potential adult resting sites and breeding habitats are abundant in the Polpithigama, Maho and Galgamuwa MOH areas. Therefore, vector control activities targeting both adult and immature stages of sand flies are recommended.

Project description:Leptospirosis is considered a neglected tropical disease despite its considerable mortality and morbidity. Lack of prediction remains a major reason for underestimating the disease. Although many models have been developed, most of them focused on the districts situated in the wet zone due to higher case numbers in that region. However, leptospirosis remains a major disease even in the dry zone of Sri Lanka. The objective of this study is to develop a time series model to predict leptospirosis in the Anuradhapura district situated in the dry zone of Sri Lanka. Time series data on monthly leptospirosis incidences from January 2008 to December 2018 and monthly rainfall, rainy days, temperature, and relative humidity were considered in model fitting. The first 72 months (55%) were used to fit the model, and the subsequent 60 months(45%) were used to validate the model. The log-transformed dependent variable was employed for fitting the Univariate seasonal ARIMA model. Based on the stationarity of the mean of the five variables, the ARDL model was selected as the multivariate time series technique. Residuals analysis was performed on normality, heteroskedasticity, and serial correlation to validate the model. The lowest AIC and MAPE were used to select the best model. Univariate models could not be fitted without adjusting the outliers. Adjusting seasonal outliers yielded better results than the models without adjustments. Best fitted Univariate model was ARIMA(1,0,0)(0,1,1)12,(AIC-1.08, MAPE-19.8). Best fitted ARDL model was ARDL(1, 3, 2, 1, 0),(AIC-2.04,MAPE-30.4). The number of patients reported in the previous month, rainfall, rainy days, and temperature showed a positive association, while relative humidity was negatively associated with leptospirosis. Multivariate models fitted better than univariate models for the original data. Best-fitted models indicate the necessity of including other explanatory variables such as patient, host, and epidemiological factors to yield better results.

Dataset Information

Naturally Occurring Microbiota Associated with Mosquito Breeding Habitats and Potential Parasitic Species against Mosquito Larvae: A Study from Gampaha District, Sri Lanka.

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Naturally Occurring Microbiota Associated with Mosquito Breeding Habitats and Potential Parasitic Species against Mosquito Larvae: A Study from Gampaha District, Sri Lanka.

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