Assessing dengue transmission risk and a vector control intervention using entomological and immunological indices in Thailand: study protocol for a cluster-randomized controlled trial.

BACKGROUND: Dengue fever is the most common and widespread mosquito-borne arboviral disease in the world. There is a compelling need for cost-effective approaches and practical tools that can reliably measure real-time dengue transmission dynamics that enable more accurate and useful predictions of incidence and outbreaks. Sensitive surveillance tools do not exist today, and only a small handful of new control strategies are available. Vector control remains at the forefront for combating dengue transmission. However, the effectiveness of many current vector control interventions is fraught with inherent weaknesses. No single vector control method is effective enough to control both vector populations and disease transmission. Evaluations of novel larval and adult control interventions are needed. METHODS/DESIGN: A cluster-randomized controlled trial will be carried out between 2017 and 2019 in urban community clusters in Khon Kaen and Roi Et cities, northeastern Thailand. The effectiveness of a pyriproxyfen/spinosad combination treatment of permanent water storage containers will be evaluated on epidemiological and entomological outcomes, including dengue incidence, number of female adult dengue vectors infected or not infected with dengue virus (DENV), human exposure to Aedes mosquito bites, and several other indices. These indices will also be used to develop predictive models for dengue transmission and impending outbreaks. Epidemiological and entomological data will be collected continuously for 2 years, with the intervention implemented after 1 year. DISCUSSION: The aims of the trial are to simultaneously evaluate the efficacy of an innovative dengue vector control intervention and developing predictive dengue models. Assessment of human exposure to mosquito bites by detecting antibodies generated against Aedes saliva proteins in human blood samples has, so far, not been applied in dengue epidemiological risk assessment and disease surveillance methodologies. Likewise, DENV detection in mosquitoes (adult and immature stages) has not been used in any practical way for routine disease surveillance strategies. The integration of multiple outcome measures will assist health authorities to better predict outbreaks for planning and applying focal and timely interventions. The trial outcomes will not only be important for Thailand, but also for the entire Southeast Asian region and further afield. TRIAL REGISTRATION: ISRCTN, ISRCTN73606171 . Registered on 23 June 2017

Characterization and Involvement of Exosomes Originating from Chikungunya Virus-Infected Epithelial Cells in the Transmission of Infectious Viral Elements

The Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that affects the world's popula-tion with chikungunya disease. Adaptation of the viral life cycle to their host cells' environment is a key step for establishing their infection and pathogenesis. Recently, the accumulating evidence advocates a principal role of extracellular vesicles (EVs), including exosomes, in both the infection and pathogenesis of infectious diseases. However, the participation of exosomes in CHIKV infec-tion and transmission is not well clarified. Here, we demonstrated that the CHIKV RNA and pro-teins were captured in exosomes, which were released by viral-infected epithelial cells. A viral genomic element in the isolated exosomes was infectious to naïve mammalian epithelial cells. The assay of particle size distribution and transmission electron microscopy (TEM) revealed CHIKV-derived exosomes with a size range from 50 to 250 nm. Treatments with RNase A, Triton X-100, and immunoglobulin G antibodies from CHIKV-positive patient plasma indicated that in-fectious viral elements are encompassed inside the exosomes. Interestingly, our viral plaque for-mation also exhibited that infectious viral elements might be securely transmitted to neighboring cells by a secreted exosomal pathway. Taken together, our recent findings emphasize the evidence for a complementary means of CHIKV infection and suggest the role of exosome-mediated CHIKV transmission

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

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

Acute Systemic Infection with Dengue Virus Leads to Vascular Leakage and Death through Tumor Necrosis Factor-α and Tie2/Angiopoietin Signaling in Mice Lacking Type I and II Interferon Receptors.

Severe dengue is caused by host responses to viral infection, but the pathogenesis remains unknown. This is, in part, due to the lack of suitable animal models. Here, we report a non-mouse-adapted low-passage DENV-3 clinical isolate, DV3P12/08, derived from recently infected patients. DV3P12/08 caused a lethal systemic infection in type I and II IFN receptor KO mice (IFN-α/β/γR KO mice), which have the C57/BL6 background. Infection with DV3P12/08 induced a cytokine storm, resulting in severe vascular leakage (mainly in the liver, kidney and intestine) and organ damage, leading to extensive hemorrhage and rapid death. DV3P12/08 infection triggered the release of large amounts of TNF-α, IL-6, and MCP-1. Treatment with a neutralizing anti-TNF-α antibody (Ab) extended survival and reduced liver damage without affecting virus production. Anti-IL-6 neutralizing Ab partly prolonged mouse survival. The anti-TNF-α Ab suppressed IL-6, MCP-1, and IFN-γ levels, suggesting that the severe response to infection was triggered by TNF-α. High levels of TNF-α mRNA were expressed in the liver and kidneys, but not in the small intestine, of infected mice. Conversely, high levels of IL-6 mRNA were expressed in the intestine. Importantly, treatment with Angiopoietin-1, which is known to stabilize blood vessels, prolonged the survival of DV3P12/08-infected mice. Taken together, the results suggest that an increased level of TNF-α together with concomitant upregulation of Tie2/Angiopoietin signaling have critical roles in severe dengue infection

Interepidemic Detection of Chikungunya Virus Infection and Transmission in Northeastern Thailand

publishedVersio

Development of eco-friendly antifungal and antibacterial adhesive derived from modified cassava starch waste/polyvinyl alcohol containing green synthesized nano-silver

Abstract Environmentally friendly biopolymer-based wood adhesives are an inevitable trend of wood product development to replace the use of harmful formaldehyde-based adhesives. In this research, a new eco-friendly modified cassava starch waste-based adhesive via carboxymethylation (CMS), and blending with polyvinyl alcohol (PVA), tannic acid (TA) and green synthesized silver nanoparticles (AgNPs) was prepared. The effects of TA content on green synthesis of AgNPs (Ag-TA) and bio-adhesive nanocomposite properties were investigated. The use of 5 wt% TA for AgNPs synthesis (Ag-TA-5) resulted in a uniform particle size distribution. The plywood prepared with Ag-TA-5 provided the highest dry and wet shear strength at 1.95 ± 0.11 MPa and 1.38 ± 0.3 MPa, respectively. The water absorption and thickness swelling of this plywood remarkably decreased up to 10.99% and 6.79%, respectively. More importantly, the presence of Ag-TA in CMS/PVA adhesive successfully inhibited the invasion of mold and bacteria. Based on the cyclic delamination test, the adhesive bond durability of bio-adhesive containing Ag-TA-5 could meet the requirement of the AITC Test T110-2007 and was comparable to commercial adhesives. The added advantage of the prepared bio-adhesive was its synthesis from agro-waste products and possible economically viable production at industrial level

Analysis of blood samples obtained from IFN-α/β/γR KO mice infected with DV3P12/08.

<p>Blood was harvested from mice infected with PBS (mock) (n = 5) or 2 × 10⁶ focus-forming units (FFU) of DV3P12/08 (n = 6) at Days 6. p.i and the number of platelets was counted by using Celltac a (Nihon Kohden). <i>P</i> = 0.04.</p

Challenge of DENV clinical isolates in IFN-α/β/γR KO mice.

<p>Challenge of DENV clinical isolates in IFN-α/β/γR KO mice.</p

Serum alanine (AST) and aspartate (ALT) transaminase levels in DV3P12/08-infected IFN-α/β/γR KO mice.

<p>Mice were intraperitoneally infected with 2 × 10⁶ focus-forming units (FFU) of DV3P12/08 or with PBS (mock) and blood samples were taken at Day 5 p.i. The levels of AST and ALT were measured using Transaminase CII-Test Wako. The results are expressed as the mean + SD of 5–6 mice per group. *<i>P</i> < 0.05 and **<i>P</i> < 0.01.</p