Dengue Virus Transmission Research Articles

Association of Dengue Disease with ABO RH Blood Group

Background: Dengue viral infection is a global public health issue. Several viral and other host factors are the contributed factors including the ABO blood group for transmission of dengue virus (DENV). ABO blood groups have been shown to involve in the intensity of DENV. This study aims to find the association ABO and Rh blood group with DENV infection. Methods: This cross-sectional study was conducted in tertiary care hospital. All DENV NS-1 positive patients were included irrespective of age and gender. The blood group of all DENV-infected patients was determined through ABO and Rh anti-sera (forward blooding grouping). Descriptive data were analyzed through SPSS and Microsoft Excel. Results: Among a total of 105 dengue-infected patients, male were 68 patients and female were 37 patients. The highest number of cases were found in the age group 21-30 years and 11-20 years. The predominant blood group was found A positive in dengue viral infected patients while AB negative blood group was found common in Rh negative blood groups. Conclusion: Blood group A positive and AB negative patients in Rh positive and Rh negative blood groups had more risk of dengue viral infection as compared to other blood groups. Further studies are necessary to find the severity of patients with different blood groups. Based on their differential distribution in the community and their correlation with mosquito performance, it can be deduced that the risk of contracting dengue infection differs with the ABO and Rh blood groups. Additional study is necessary to determine whether HLA, dengue serotype, and ABO are experimental/predictor factors, and whether certain blood subgroups are associated with a disproportionately higher risk of dengue virus infection. Keywords: Dengue viral infection, Serotype, Anti-A, Anti-B, Anti-D, ABO, Rh blood group

Detection of Dengue Virus 2 with Single Infected Mosquito Resolution Using Yeast Affinity Bionanofragments and Plasmonic SERS Nanoboxes.

Dengue disease is an emerging global threat triggered by dengue virus (DENV) transmission, primarily by the mosquito Aedes aegypti. The accurate surveillance and sensitive detection of DENV in mosquito populations are critical for the protection of human populations worldwide that are in the habitat of these mosquito species. There are four DENV serotypes with DENV2 reported to cause the most severe complications. There are limited ultrasensitive methods to early detect DENV2 mosquito infection and prevent human infection. Herein, we report an innovative nanobased immunoassay platform for early, specific, and ultrasensitive detection of DENV2-secreted nonstructural 1 (NS1) protein biomarker in single infected mosquitoes with the limit of detection of 500 fg of recombinant DENV2 NS1. The high sensitivity and DENV2 serotype specificity of the platform are the result of using nanomixing, plasmonic SERS nanoboxes, and yeast affinity bionanofragments displaying single-chain variable fragments (nanoyeast scFvs). Nanoyeast scFvs used for high affinity capture of DENV2 NS1 provided an innovative and cost-efficient alternative to monoclonal antibodies and differentiated DENV2 NS1 from other DENV serotypes and Zika virus NS1. The platform used electrohydrodynamically driven nanomixing to enhance NS1 capture by the nanoyeast scFvs while reducing nonspecific interactions. High sensitivity detection of captured DENV2 NS1 was achieved using NS1-specific surface-enhanced Raman scattering (SERS) nanotags. These nanotechnologies provide a significant innovation for early DENV2 detection in single infected mosquitoes, improving the accurate surveillance of mosquito habitats and preventing infection and severe disease arising from DENV2 transmission.

Dengue Transmission From Donor to Recipient After Deceased Donor Liver Transplant; Report of a Case and Review of Literature: Is Liver a Privileged Organ?

Nonvector transmission of dengue virus via organ transplantis rare.We report a probable case of dengue virus transmission via deceased donorlivertransplant. The donor was asymptomatic for dengue infection, and the liver recipient developed fever and thrombocytopenia 8 days after transplant. The recipient's testresults were positive forthe antigen for nonstructural protein 1 and for immunoglobulin M; however, donor serum samples were not available. Other transplant recipients (renal allograft) also showed dengue-positive test results during the same period, and these patients eventually died. Dengue illness in the liver recipient was severe and associated with shock and cardiomyopathy that required mechanical ventilation, intensive care, and cessation of immunosuppression; however, the patient subsequently improved and was discharged. This case highlights the importance of a high index of suspicion for dengue infection in the presence of graft dysfunction in dengue-endemic areas like India, as well as the importance of the test for the nonstructural protein 1 antigen during dengue outbreaks. These considerations should be included on the standard evaluation of solid-organ transplant donors.

Contribution of phylogenetics to understanding the evolution and epidemiology of dengue virus.

Dengue virus (DENV) is one of the most important arboviral pathogens in the tropics and subtropics, and nearly one‐third of the world's population is at risk of infection. The transmission of DENV involves a sylvatic cycle between nonhuman primates (NHP) and Aedes genus mosquitoes, and an endemic cycle between human hosts and predominantly Aedes aegypti. DENV belongs to the genus Flavivirus of the family Flaviviridae and consists of four antigenically distinct serotypes (DENV‐1‐4). Phylogenetic analyses of DENV have revealed its origin, epidemiology, and the drivers that determine its molecular evolution in nature. This review discusses how phylogenetic research has improved our understanding of DENV evolution and how it affects viral ecology and improved our ability to analyze and predict future DENV emergence.

Study of Dengue Virus Transovarial Transmission in Aedes spp. in Ternate City Using Streptavidin-Biotin-Peroxidase Complex Immunohistochemistry.

Aedes aegypti is the most dominant vector in the transmission of dengue hemorrhagic fever (DHF). In addition to Ae. aegypti, Ae. albopictus is a secondary vector of the dengue virus, and both species are widespread in Indonesia. The dengue virus is transmitted from person to person through the bite of an Aedes spp. The vertical (transovarial) transmission of the dengue virus from infective female mosquitoes to their offspring is one of the means by which the dengue virus maintains its existence in nature. Transovarial dengue virus transmission in Aedes spp. mosquitoes contributes to the spread and maintenance of the dengue epidemic. This study employed a qualitative survey to detect dengue virus transovarial transmission in Ternate using the streptavidin-biotin-peroxidase complex (ISBPC) immunohistochemical test. The ISBPC examination of samples collected from the four subdistricts in Ternate revealed a positive result for transovarial transmission of dengue virus. Four Aedes spp., including two Ae. aegypti females, one Ae. albopictus female, and one Ae. albopictus male, tested positive for transovarial transmission of dengue virus in the district of North Ternate. Four Aedes spp., including three Ae. aegypti females and one Ae. aegypti male, were found to be positive for the transovarial transmission of dengue virus in the Central Ternate district. Seven Aedes spp., including five Ae. aegypti females, one Ae. aegypti male, and one Ae. albopictus female, tested positive for transovarial transmission of the dengue virus in the district of South Ternate city. One Ae. aegypti male showed positive results for transovarial transmission of dengue virus in the Ternate Island District. In this study, the transovarial transmission of the dengue virus occurred in both Aedes spp. female and male mosquitoes. It was demonstrated that Aedes spp. carry the dengue virus in their ovaries and can pass it on to their offspring. As a result, the cycle of passing the dengue virus on to local mosquito populations in the city of Ternate is not going to end just yet.

Characterization of dengue virus 3'UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva.

Dengue viruses (DENV) are expanding global pathogens that are transmitted through the bite of mosquitoes, mostly Aedes aegypti. As RNA viruses, DENV rely on RNA-binding proteins (RBPs) to complete their life cycle. Alternatively, RBPs can act as restriction factors that prevent DENV multiplication. While the importance of RBPs is well-supported in humans, there is a dearth of information about their influence on DENV transmission by mosquitoes. Such knowledge could be harnessed to design novel, effective interventions against DENV. Here, we successfully adapted RNA-affinity chromatography coupled with mass spectrometry-a technique initially developed in mammalian cells-to identify RBPs in Ae. aegypti cells. We identified fourteen RBPs interacting with DENV serotype 2 3'UTR, which is involved in the viral multiplication and produces subgenomic flaviviral RNA (sfRNA). We validated the RNA affinity results for two RBPs by confirming that AePur binds the 3'UTR, whereas AeStaufen interacts with both 3'UTR and sfRNA. Using in vivo functional evaluation, we determined that RBPs like AeRan, AeExoRNase, and AeRNase have pro-viral functions, whereas AeGTPase, AeAtu, and AePur have anti-viral functions in mosquitoes. Furthermore, we showed that human and mosquito Pur homologs have a shared affinity to DENV2 RNA, although the anti-viral effect is specific to the mosquito protein. Importantly, we revealed that AeStaufen mediates a reduction of gRNA and sfRNA copies in several mosquito tissues, including the salivary glands and that AeStaufen-mediated sfRNA reduction diminishes the concentration of transmission-enhancing sfRNA in saliva, thereby revealing AeStaufen's role in DENV transmission. By characterizing the first RBPs that associate with DENV2 3'UTR in mosquitoes, our study unravels new pro- and anti-viral targets for the design of novel therapeutic interventions as well as provides foundation for studying the role of RBPs in virus-vector interactions.

Spatial and Temporal Distribution of Aedes aegypti and Aedes albopictus Oviposition on the Coast of Paraná, Brazil, a Recent Area of Dengue Virus Transmission.

Aedes aegypti and Aedes albopictus are considered the most important vectors of arboviruses in the world. Aedes aegypti is the primary vector of dengue, urban yellow fever, chikungunya and zika in Brazil, and Ae. albopictus is considered a potential vector. Distribution patterns and the influence of climatic variables on the oviposition of Ae. aegypti and Ae. albopictus were evaluated in Morretes, a tourist city in the coastal area of Paraná State, Brazil, which has recently been experiencing cases of dengue fever. Eggs were collected using ovitraps over a period of one year (September 2017 to September 2018) and reared from hatching until the emergence of the adults. Both Aedes species were found in anthropized areas with a high human density index. Findings suggest that the monthly average temperature (LRT = 16.65, p = 0.001) had significant positive influences on the oviposition of the Aedes species. Considering the wide distribution of DENV around the Paraná coast and the presence of Ae. albopictus alongside Ae. aegypti, studies on natural arbovirus infection patterns and seasonality are recommended in the region.

Prevalence and resting behaviour of dengue vectors, Aedes aegypti and Aedes albopictus in dengue high risk urban settings in Colombo, Sri Lanka

• Aedes aegypti is the predominant vector in urban areas of the Colombo district. • Ae. aegypti adult density correlates with climatic factors such as monthly rainfall and mean temperature. • Ae. aegypti adult density positively correlates with the one-month lagged dengue case number. • Ae. aegypti preferred indoor resting areas in urban areas were bedrooms and living rooms. Understanding local geographical variation in vector density and bionomics related to virus transmission are critical for planning effective vector control programs to control dengue virus transmission. This study investigated the prevalence and resting behaviour of Aedes aegypti and Ae. albopictus in three dengue high-risk areas in the Colombo District, Sri Lanka. Monthly sampling of resting adult mosquitoes was conducted from August 2019 to February 2020. Thirty-seven percent of the households (289/776) harboured Aedes mosquitoes, and 603 Aedes mosquitoes were collected. The proportion of Ae. aegypti was higher in the overall collection during the collection period (94% [569/603]), and 62% (352/569) were females. Significant monthly variations in Ae. aegypti were observed with respect to the indices: number of females per surveyed house (F/SH; p = 0.001), number of females per Aedes positive house (F/PH; p = 0.029), adult house index (AHI; p = 0.001), adult density (AD; p = 0.005) and resting ratio (RR; p = 0.001). AHI, AD, and RR had statistically significant positive correlations with monthly rainfall (p = 0.001, p = 0.011, and p = 0.002 respectively) and one-month lagged dengue cases (p = 0.002, p = 0.005, and p < 0.001 respectively). A statistically significantly higher proportion of Ae. aegypti females were caught resting indoors (N = 309, 88%) than outdoors (N = 43, 12%; p < 0.001). The most common resting areas were bedrooms (51%) & living/dining rooms (37%), and places were under or on furniture (47%) & hangings (34%) for Ae. aegypti . Conversely, 74% of female Ae. albopictus were collected outdoors. Results of this study could have strong implications to improve vector surveillance and control by early detection of dengue to detect outbreaks and minimization of disease transmission.

Dengue virus among HIV-infected pregnant women attending antenatal care in Luanda, Angola: An emerging public health concern

The dissemination of the dengue virus (DENV) in endemic regions with HIV is a public health concern with greater importance when there is evidence of vertical transmission of DENV during pregnancy. Herein, we investigated DENV among HIV-infected pregnant women in Luanda, the capital city of Angola. This was part of a cross-sectional study carried out on 42 pregnant women newly diagnosed with HIV. A total of 36 plasma samples from the 42 HIV-positive pregnant women were screened for DENV using RT‐PCR and ELISA. None of the specimens tested positive for DENV by RT-PCR. Regarding seroprevalence, 94.4% of the samples were positive for IgG and 11.1% for IgM. Recent infection (IgG-/IgM+ or IgG+/IgM+) was detected in 11.1% of the samples and past infection (IgG+/IgM-) in 83.3%. The risk of recent infection was higher in pregnant women over 25 years of age [OR: 13.0 (95% CI: 1.14–148), p = 0.039]. Our study showed laboratory evidence of a recent DENV infection among HIV-infected pregnant women attending antenatal care in Luanda. Our findings provide critical data regarding DENV infection among HIV-infected pregnant women in Luanda. Future studies involving a larger sample size of HIV-infected pregnant women are necessary to support ongoing public health programs to combat arboviruses in Angola.

Ingestion of spinosad-containing toxic sugar bait alters Aedes albopictus vector competence and vectorial capacity for dengue virus.

Dengue virus (DENV) is a highly prevalent vector-borne virus that causes life-threatening illnesses to humans worldwide. The development of a tool to control vector populations has the potential to reduce the burden of DENV. Toxic sugar bait (TSB) provides a form of vector control that takes advantage of the sugar-feeding behavior of adult mosquitoes. However, studies on the effect of ingestion of toxins in TSB on vector competence and vectorial capacity for viruses are lacking. This study evaluated vector competence for DENV serotype-1 of Aedes albopictus at 7 and 14 days post-ingestion of TSB formulated with spinosad (of bacteria origin) as an oral toxin. Our results and others were modeled to estimate effects on Ae. albopictus vectorial capacity for DENV. Ingestion of TSB caused a reduction in survival of females, but increased mosquito susceptibility to DENV infection, disseminated infection, and transmission. However, this increase in vector competence was obviated by the reduction in survival, leading to a lower predicted vectorial capacity. The findings of this study highlight the importance of evaluating the net impact of TSB ingestion on epidemiological parameters of vectorial capacity in the context of vector control efforts to reduce the risk of transmission of vector-borne viruses.

Studies on the fitness characteristics of wMel- and wAlbB-introgressed Aedes aegypti (Pud) lines in comparison with wMel- and wAlbB-transinfected Aedes aegypti (Aus) and wild-type Aedes aegypti (Pud) lines.

Wolbachia, an intracellular maternally transmitted endosymbiont, has been shown to interfere with the replication of dengue virus in Aedes aegypti mosquitoes. The Wolbachia-transinfected Ae. aegypti has been currently released in many countries to test its effectiveness in preventing the transmission of dengue virus. ICMR-Vector Control Research Centre in collaboration with World Mosquito Program Monash University, Australia, has generated two new Wolbachia-introgressed Ae. aegypti Puducherry (Pud) lines via backcrossing Ae. aegypti females of Australian (Aus) strains, infected with wMel and wAlbB Wolbachia with wild-type Ae. aegypti Puducherry (Pud) males. Wolbachia infections are known to induce a fitness cost and confer benefit on the host mosquito populations that will influence spread of the Wolbachia into native wild mosquito populations during the field release. Hence, the induced fitness cost or benefit/advantage in the two newly generated Ae. aegypti (Pud) lines was assessed in the laboratory in comparison with the wild-type Ae. aegypti (Pud) strain. In addition, maternal transmission (MT) efficiency, induced cytoplasmic incompatibility (CI), and insecticide resistance status of the two (Pud) lines were determined to assess the likely frequency of wMel and wAlbB infections in the native wild population after field invasion. The study shows that wMel and wAlbB infections did not induce any fitness cost on the two newly generated (Pud) lines. Rather, in terms of wing length, fecundity, egg hatch rate, and adult survival, the Wolbachia introgression conferred fitness benefits on the (Pud) lines compared to uninfected Wolbachia free wild Ae. aegypti population. wMel and wAlbB exhibited a high maternal transmission (99–100%) and induced nearly complete (98–100%) cytoplasmic incompatibility. Both the (Pud) lines were resistant to deltamethrin, malathion, DDT, and temephos, and the level of resistance was almost the same between the two lines as in the wild type. Overall, the stable association of wMel and wAlbB established with Ae. aegypti and the reproductive advantages of the (Pud) lines encourage a pilot release in the field for population replacement potential.

Modeling optimal vaccination strategy for dengue epidemic model: a case study of India

Dengue fever is a mosquito-borne infectious disease which is transmitted through Aedes aegypti mosquitoes. It is one of the major global health issues in the tropical and subtropical regions of the world. Its dynamics are very complicated owing to the coupling among multiple transmission pathways and various components in pathogen. Due to absence of proper medicine or vaccination, mathematical modeling plays an important role in understanding the disease dynamics and in designing strategies to control the spread of dengue virus. In this paper, we studied a non-linear vector-host model to investigate the transmission dynamics of dengue virus which can be controlled by vaccination as well as treatment. Analysis of model start with the basic reproduction number , when it is less than one, the system become locally asymptotically stable about the virus-free equilibrium point. We calibrated our proposed model corresponding to transmission of dengue virus using real data for six Indian states, namely Tamil Nadu, Kerala, Delhi, Gujarat, Rajasthan and Karnataka by using the least square method. Moreover, we performed normalized forward sensitivity analysis of the basic reproduction number and obtained that mitigating the disease transmission rate of mosquito and human population is the most important factor in achieving dengue control. Finally, an objective functional has been developed to minimize the cost of the vaccination and solved with the aid of the Pontryagin’s Maximum Principle. The implementation of the optimal treatment policy shows a significant reduction of the hospitalized individuals as well as infected individuals. We then performed extensive numerical simulations to validate our theoretical analysis with aid of the estimated model parameters.

Potential Of Transmission Of The Dengue Virus Based On Entomological Index And Maya Index In Kalumata And North Mangga Dua villages,Ternate City

Dengue hemorrhagic fever (DHF) is the result of the interaction between the dengue virus, humans and environmental factors. Dengue fever can be spread through the bite of the Aedes aegypti mosquito. Knowledge related to larval habitat is very important to limit the spread of Dengue virus by reducing the population level of Ae. aegypti. This study aims to determine the potential for dengue virus transmission based on the Entomological Index and Maya Index. The study was conducted in 2 villages in Ternate City for 1 month. To calculate the entomological index value, it is done by observing the characteristics of the habitat, namely by observing the container that is the habitat of Ae. aegypti. The calculated parameters are Container Index (CI), Breteau Index (BI), House Index (HI), Density Figure (DF), and calculate the BRI and HRI values to determine the virtual index. Based on the results of the study, it is known that the entomological index value of Kalumata and North Mangga Dua villages is CI 20.71%, HI 84.0% and BI 361%, with the average DF being in the high category (DF = 7 and 9). The virtual nidex value is in the medium category, namely MI = 55%, with a large BRI value = 56% (high category) and HRI = 45.5% (medium category), meaning that Kalumata and North Mangga Dua villages have the potential for virus transmission to occur Dengue.

First Report of Aedes vittatus in Islamabad, Pakistan.

Aedes vittatus is distributed throughout Asia, Africa, and Europe and can transmit dengue, chikungunya, yellow fever, and Zika viruses. Like other Aedes species, larvae develop in both natural and artificial containers in urban, suburban, and rural areas. In September 2021, an entomological survey was conducted at the National Institute of Health of Pakistan (NIH) and adjacent housing within the NIH colony. All containers with water were examined for Aedes mosquitoes at 150 locations, including residential properties, a plant nursery, junkyards, and recreational parks and playgrounds. A total of 103 larvae, 37 pupae, 5 female and 2 male Ae. vittatus were collected from a fountain. This was the first detection of Ae. vittatus in urban Islamabad. Additional vector surveillance is needed to better understand the geographical distribution, ecology, and behavior of this invasive species and to understand its possible role in the transmission of dengue and chikungunya viruses in Pakistan.

A multiple-target mRNA-LNP vaccine induces protective immunity against experimental multi-serotype DENV in mice.

Dengue virus (DENV) is a mosquito-borne virus with a rapid spread to humans, causing mild to potentially fatal illness in hundreds of millions of people each year. Due to the large number of serotypes of the virus, there remains an unmet need to develop protective vaccines for a broad spectrum of the virus. Here, we constructed a modified mRNA vaccine containing envelope domain III (E-DIII) and non-structural protein 1 (NS1) coated with lipid nanoparticles. This multi-target vaccine induced a robust antiviral immune response and increased neutralizing antibody titers that blocked all four types of DENV infection in vitro without significant antibody-dependent enhancement (ADE). In addition, there was more bias for Th1 than Th2 in the exact E-DIII and NS1-specific T cell responses after a single injection. Importantly, intramuscular immunization limited DENV transmission in vivo and eliminated vascular leakage. Our findings highlight that chimeric allogeneic structural and non-structural proteins can be effective targets for DENV vaccine and that they can prevent the further development of congenital DENV syndrome.

Expression of fatty acid synthase genes and their role in development and arboviral infection of Aedes aegypti

BackgroundFatty acids are the building blocks of complex lipids essential for living organisms. In mosquitoes, fatty acids are involved in cell membrane production, energy conservation and expenditure, innate immunity, development and reproduction. Fatty acids are synthesized by a multifunctional enzyme complex called fatty acid synthase (FAS). Several paralogues of FAS were found in the Aedes aegypti mosquito. However, the molecular characteristics and expression of some of these paralogues have not been investigated.MethodsGenome assemblies of Ae. aegypti were analyzed, and orthologues of human FAS was identified. Phylogenetic analysis and in silico molecular characterization were performed to identify the functional domains of the Ae. aegypti FAS (AaFAS). Quantitative analysis and loss-of-function experiments were performed to determine the significance of different AaFAS transcripts in various stages of development, expression following different diets and the impact of AaFAS on dengue virus, serotype 2 (DENV2) infection and transmission.ResultsWe identified seven putative FAS genes in the Ae. aegypti genome assembly, based on nucleotide similarity to the FAS proteins (tBLASTn) of humans, other mosquitoes and invertebrates. Bioinformatics and molecular analyses suggested that only five of the AaFAS genes produce mRNA and therefore represent complete gene models. Expression levels of AaFAS varied among developmental stages and between male and female Ae. aegypti. Quantitative analyses revealed that expression of AaFAS1, the putative orthologue of the human FAS, was highest in adult females. Transient knockdown (KD) of AaFAS1 did not induce a complete compensation by other AaFAS genes but limited DENV2 infection of Aag2 cells in culture and the midgut of the mosquito.ConclusionAaFAS1 is the predominant AaFAS in adult mosquitoes. It has the highest amino acid similarity to human FAS and contains all enzymatic domains typical of human FAS. AaFAS1 also facilitated DENV2 replication in both cell culture and in mosquito midguts. Our data suggest that AaFAS1 may play a role in transmission of dengue viruses and could represent a target for intervention strategies.Graphical

An ensemble forecast system for tracking dynamics of dengue outbreaks and its validation in China.

As a common vector-borne disease, dengue fever remains challenging to predict due to large variations in epidemic size across seasons driven by a number of factors including population susceptibility, mosquito density, meteorological conditions, geographical factors, and human mobility. An ensemble forecast system for dengue fever is first proposed that addresses the difficulty of predicting outbreaks with drastically different scales. The ensemble forecast system based on a susceptible-infected-recovered (SIR) type of compartmental model coupled with a data assimilation method called the ensemble adjusted Kalman filter (EAKF) is constructed to generate real-time forecasts of dengue fever spread dynamics. The model was informed by meteorological and mosquito density information to depict the transmission of dengue virus among human and mosquito populations, and generate predictions. To account for the dramatic variations of outbreak size in different seasons, the effective population size parameter that is sequentially updated to adjust the predicted outbreak scale is introduced into the model. Before optimizing the transmission model, we update the effective population size using the most recent observations and historical records so that the predicted outbreak size is dynamically adjusted. In the retrospective forecast of dengue outbreaks in Guangzhou, China during the 2011–2017 seasons, the proposed forecast model generates accurate projections of peak timing, peak intensity, and total incidence, outperforming a generalized additive model approach. The ensemble forecast system can be operated in real-time and inform control planning to reduce the burden of dengue fever.

Effective Infection with Dengue Virus in Experimental Neonate and Adult Mice through the Intranasal Route.

Dengue virus, the causative agent of dengue fever, life-threatening hemorrhagic fever, and shock syndrome, is mainly transmitted to humans through mosquito vectors. It can also be transmitted through atypical routes, including needle stick injury, vertical transmission, blood transfusion, and organ transplantation. In addition, sporadic cases which have no clear infectious causes have raised the respiratory exposure concerns, and the risks remain unclear. Here, we analyze the respiratory infectivity of the dengue virus in BALB/c suckling and adult immunodeficient mice by the intranasal inoculation of dengue virus serotype 2. The infected mice presented with clinical symptoms, including excitement, emaciation, malaise, and death. Viremia was detected for 3 days post inoculation. Histopathological changes were observed in the brain, liver, and spleen. The virus showed evident brain tropism post inoculation and viral loads peaked at 7 days post inoculation. Furthermore, the virus was isolated from the infected mice; the sequence homology between the origin and isolates was 99.99%. Similar results were observed in adult IFN-α/β receptor-deficient mice. Overall, dengue virus can infect suckling mice and adult immune-deficient mice via the nasal route. This study broadens our perception of atypical dengue transmission routes and provides evidence of nasal transmission of dengue virus in the absence of mosquito vectors.

Disruption of spatiotemporal clustering in dengue cases by wMel Wolbachia in Yogyakarta, Indonesia

Dengue exhibits focal clustering in households and neighborhoods, driven by local mosquito population dynamics, human population immunity, and fine scale human and mosquito movement. We tested the hypothesis that spatiotemporal clustering of homotypic dengue cases is disrupted by introduction of the arbovirus-blocking bacterium Wolbachia (wMel-strain) into the Aedes aegypti mosquito population. We analysed 318 serotyped and geolocated dengue cases (and 5921 test-negative controls) from a randomized controlled trial in Yogyakarta, Indonesia of wMel deployments. We find evidence of spatial clustering up to 300 m among the 265 dengue cases (3083 controls) in the untreated trial arm. Participant pairs enrolled within 30 days and 50 m had a 4.7-fold increase (compared to 95% CI on permutation-based null distribution: 0.1, 1.2) in the odds of being homotypic (i.e. potentially transmission-related) as compared to pairs occurring at any distance. In contrast, we find no evidence of spatiotemporal clustering among the 53 dengue cases (2838 controls) resident in the wMel-treated arm. Introgression of wMel Wolbachia into Aedes aegypti mosquito populations interrupts focal dengue virus transmission leading to reduced case incidence; the true intervention effect may be greater than the 77% efficacy measured in the primary analysis of the Yogyakarta trial.

Detection of Dengue Virus From Aedes aegypti (Diptera, Culicidae) in Field-Caught Samples From Makkah Al-Mokarramah, Kingdom of Saudi Arabia, Using RT-PCR.

Dengue fever (DF) is endemic to Makkah and Jeddah, the Kingdom of Saudi Arabia (KSA). However, until recently, the circulation of dengue virus (DENV) in Aedes mosquitoes in these areas was unknown. Serological surveillance of DENV in Ae aegypti is a powerful tool for early detection of dengue outbreaks and essential for developing effective control strategies. Therefore, this research aimed to examine a sample of adult Ae aegypti mosquitoes from Makkah, KSA, to detect DENV. In total, 1295 Ae aegypti mosquitoes were collected from the field from target areas of Makkah with a high incidence and prevalence of DF. The samples were divided into 259 coded pools (five mosquitoes in each) and preserved in 1.5 mL plastic tubes. The tubes were labeled, capped, and stored at−86°C until use. RT-PCR was used to detect DENV in the samples. All positive pools were confirmed by RT-PCR. The RT-PCR products were analyzed by gel electrophoresis (1.5% agarose in Tris-acetate EDTA buffer), stained with ethidium bromide, and visualized. DENV was isolated from six female Ae Aegypti collected from six pools (out of 259 pools). No other viruses were detected. Only five of the nine target localities had positive pools. Samples from the remaining four localities yielded negative results. Four DENV-positive mosquitoes were collected at the aquatic stages, and two were collected at the adult stage. These results show the circulation of DENV in adult mosquitoes and offspring, indicating vertical transmission of DENV. In conclusion, this study found that, in Makkah, DENV is circulating in dengue vectors with a high significance rate, suggesting the possibility of a dengue outbreak in the future; therefore, a sensitive surveillance system is vital to predict the outbreak and for early intervention and control.