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      Dengue virus in humans and mosquitoes and their molecular characteristics in northeastern Thailand 2016-2018

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          Abstract

          Dengue is hyperendemic in most Southeast Asian countries including Thailand, where all four dengue virus serotypes (DENV-1 to -4) have circulated over different periods and regions. Despite dengue cases being annually reported in all regions of Thailand, there is limited data on the relationship of epidemic DENV infection between humans and mosquitoes, and about the dynamics of DENV during outbreaks in the northeastern region. The present study was conducted in this region to investigate the molecular epidemiology of DENV and explore the relationships of DENV infection in humans and in mosquitoes during 2016–2018. A total of 292 dengue suspected patients from 11 hospitals and 902 individual mosquitoes (at patient’s houses and neighboring houses) were recruited and investigated for DENV serotypes infection using PCR. A total of 103 patients and 149 individual mosquitoes were DENV -positive. Among patients, the predominant DENV serotypes in 2016 and 2018 were DENV-4 (74%) and DENV-3 (53%) respectively, whereas in 2017, DENV-1, -3 and -4 had similar prevalence (38%). Additionally, only 19% of DENV infections in humans and mosquitoes at surrounding houses were serotypically matched, while 81% of infections were serotypically mismatched, suggesting that mosquitoes outside the residence may be an important factor of endemic dengue transmission. Phylogenetic analyses based on envelope gene sequences showed the genotype I of both DENV-1 and DENV-4, and co-circulation of the Cosmopolitan and Asian I genotypes of DENV-2. These strains were closely related to concurrent strains in other parts of Thailand and also similar to strains in previous epidemiological profiles in Thailand and elsewhere in Southeast Asia. These findings highlight genomic data of DENV in this region and suggest that people’s movement in urban environments may result in mosquitoes far away from the residential area being key determinants of DENV epidemic dynamics.

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          Dengue, Urbanization and Globalization: The Unholy Trinity of the 21st Century

          Dengue is the most important arboviral disease of humans with over half of the world’s population living in areas of risk. The frequency and magnitude of epidemic dengue have increased dramatically in the past 40 years as the viruses and the mosquito vectors have both expanded geographically in the tropical regions of the world. There are many factors that have contributed to this emergence of epidemic dengue, but only three have been the principal drivers: 1) urbanization, 2) globalization and 3) lack of effective mosquito control. The dengue viruses have fully adapted to a human-Aedes aegypti-human transmission cycle, in the large urban centers of the tropics, where crowded human populations live in intimate association with equally large mosquito populations. This setting provides the ideal home for maintenance of the viruses and the periodic generation of epidemic strains. These cities all have modern airports through which 10s of millions of passengers pass each year, providing the ideal mechanism for transportation of viruses to new cities, regions and continents where there is little or no effective mosquito control. The result is epidemic dengue. This paper discusses this unholy trinity of drivers, along with disease burden, prevention and control and prospects for the future.
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            A new, cost-effective, battery-powered aspirator for adult mosquito collections.

            We report the development of a new mosquito aspirator with the same aspiration capacity (airflow) of the CDC Backpack Aspirator (CDC-BP), but smaller and lighter (0.8 kg without battery), less expensive (US$45-70), easier to build, and compatible with the use of telescoping extension poles to access hard-to-reach locations. The performance of this new aspirator, named "Prokopack," was compared with the CDC-BP in laboratory settings as well as in paired collections in combined sewer overflow (CSO) tunnels in Atlanta, GA, and indoor mosquito collections in Iquitos, Peru. The difference in suction power between both aspirators (average, 0.29-0.43 m/s) was negligible. However, 2.3 times more mosquitoes were collected using the Prokopack in the upper wall (>1.5 m) and ceilings of CSO tunnels than with the CDC-BP in lower walls. Indoor collection in Iquitos yielded significantly more total mosquito numbers [including Culex pipiens complex, Culex (melanoconion) sp., and Mansonia sp.] and Aedes aegypti (L.) in the Prokopack than in the CDC-BP. Our results demonstrate the effectiveness of the Prokopack to collect different mosquito species in different epidemiological settings.
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              Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction.

              We report on the development and application of a rapid assay for detecting and typing dengue viruses. Oligonucleotide consensus primers were designed to anneal to any of the four dengue virus types and amplify a 511-bp product in a reverse transcriptase-polymerase chain reaction (PCR). First, we produced a cDNA copy of a portion of the viral genome in a reverse transcriptase reaction in the presence of primer D2 and then carried out a standard PCR (35 cycles of heat denaturation, annealing, and primer extension) with the addition of primer D1. The resulting double-stranded DNA product of the RT-PCR was typed by two methods: dot blot hybridization of the 511-bp amplified product to dengue virus type-specific probes or a second round of PCR amplification (nested PCR) with type-specific primers, yielding DNA products the unique sizes of which were diagnostic for each dengue virus serotype. The accumulated data demonstrated that dengue viruses can be accurately detected and typed from viremic human serum samples.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: MethodologyRole: Supervision
                Role: MethodologyRole: SupervisionRole: Writing – review & editing
                Role: Investigation
                Role: SupervisionRole: Writing – review & editing
                Role: Funding acquisitionRole: Supervision
                Role: Funding acquisitionRole: MethodologyRole: SupervisionRole: Writing – review & editing
                Role: Formal analysis
                Role: Formal analysisRole: Supervision
                Role: Project administration
                Role: Funding acquisitionRole: Project administrationRole: Writing – review & editing
                Role: Funding acquisitionRole: Writing – review & editing
                Role: Funding acquisitionRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Funding acquisitionRole: Project administrationRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                14 September 2021
                2021
                : 16
                : 9
                : e0257460
                Affiliations
                [1 ] Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand
                [2 ] HPV & EBV and Carcinogenesis Research Group, Khon Kaen University, Khon Kaen, Thailand
                [3 ] Mahidol-Osaka Center for Infectious Diseases, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
                [4 ] Research Institute for Microbial Diseases, Osaka University, Osaka, Japan
                [5 ] Program in Bioinformatics and Computational Biology, Graduate School, Chulalongkorn University, Bangkok, Thailand
                [6 ] Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
                [7 ] Department of Disease Control, Office of Disease Prevention and Control, Region 7 Khon Kaen, Ministry of Public Health, Khon Kaen, Thailand
                [8 ] Faculty of Science and Technology, Norwegian University of Life Sciences, Ås, Norway
                [9 ] Public Health & Malaria Control, PT Freeport Indonesia/International SOS, Kuala Kencana, Papua, Indonesia
                [10 ] Department of Entomology, Faculty of Agriculture, Kasetsart University, Bangkok, Thailand
                [11 ] MRC Tropical Epidemiology Group, London School of Hygiene and Tropical Medicine, London, United Kingdom
                Defense Threat Reduction Agency, UNITED STATES
                Author notes

                Competing Interests: The authors declare no competing interests exist.

                Author information
                https://orcid.org/0000-0003-2797-043X
                Article
                PONE-D-21-11383
                10.1371/journal.pone.0257460
                8439490
                34520486
                d5cd635f-b6c7-46aa-9de7-434c1b599c5c
                © 2021 Nonyong et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 7 April 2021
                : 1 September 2021
                Page count
                Figures: 5, Tables: 2, Pages: 19
                Funding
                Funded by: Research Council of Norway
                Award ID: 250443
                Award Recipient :
                Funded by: Invitation Research Grant
                Award ID: IN62113
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100009619, Japan Agency for Medical Research and Development;
                Award ID: JP19fm0108003 and 20wm0225010h0101
                Award Recipient :
                This work was supported by the Research Council of Norway (project no. 250443); the Invitation Research Grant, Faculty of Medicine, Khon Kaen University, Thailand, grant number IN62113; the Japan Agency for Medical Research and Development (AMED) JP19fm0108003 and 20wm0225010h0101.
                Categories
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