Mechanical transmission of dengue virus by Aedes aegypti may influence disease transmission dynamics during outbreaks.

BACKGROUND: Dengue virus outbreaks are increasing in number and severity worldwide. Viral transmission is assumed to require a minimum time period of viral replication within the mosquito midgut. It is unknown if alternative transmission periods not requiring replication are possible. METHODS: We used a mouse model of dengue virus transmission to investigate the potential of mechanical transmission of dengue virus. We investigated minimal viral titres necessary for development of symptoms in bitten mice and used resulting parameters to inform a new model of dengue virus transmission within a susceptible population. FINDINGS: Naïve mice bitten by mosquitoes immediately after they took partial blood meals from dengue infected mice showed symptoms of dengue virus, followed by mortality. Incorporation of mechanical transmission into mathematical models of dengue virus transmission suggest that this supplemental transmission route could result in larger outbreaks which peak sooner. INTERPRETATION: The potential of dengue transmission routes independent of midgut viral replication has implications for vector control strategies that target mosquito lifespan and suggest the possibility of similar mechanical transmission routes in other disease-carrying mosquitoes. FUNDING: This study was funded by grants from the National Health Research Institutes, Taiwan (04D2-MMMOST02), the Human Frontier Science Program (RGP0033/2021), the National Institutes of Health (1R01AI143698-01A1, R01AI151004 and DP2AI152071) and the Ministry of Science and Technology, Taiwan (MOST104-2321-B-400-016).

Use of unmanned ground vehicle systems in urbanized zones: A study of vector Mosquito surveillance in Kaohsiung.

Dengue fever is a vector-borne disease that has become a serious global public health problem over the past decade. An essential aspect of controlling and preventing mosquito-borne diseases is reduction of mosquito density. Through the process of urbanization, sewers (ditches) have become easy breeding sources of vector mosquitoes. In this study, we, for the first time, used unmanned ground vehicle systems (UGVs) to enter ditches in urban areas to observe vector mosquito ecology. We found traces of vector mosquitoes in ~20.7% of inspected ditches, suggesting that these constitute viable breeding sources of vector mosquitoes in urban areas. We also analyzed the average gravitrap catch of five administrative districts in Kaohsiung city from May to August 2018. The gravitrap indices of Nanzi and Fengshan districts were above the expected average (3.26), indicating that the vector mosquitoes density in these areas is high. Using the UGVs to detect positive ditches within the five districts followed by insecticide application generally yielded good control results. Further improving the high-resolution digital camera and spraying system of the UGVs may be able to effectively and instantly monitor vector mosquitoes and implement spraying controls. This approach may be suitable to solve the complex and difficult task of detecting mosquito breeding sources in urban ditches.

Comparison of Fan-Traps and Gravitraps for Aedes Mosquito Surveillance in Taiwan.

A key component of integrated vector management strategies is the efficient implementation of mosquito traps for surveillance and control. Numerous trap types have been created with distinct designs and capture mechanisms, but identification of the most effective trap type is critical for effective implementation. For dengue vector surveillance, previous studies have demonstrated that active traps utilizing CO2 attractant are more effective than passive traps for capturing Aedes mosquitoes. However, maintaining CO2 supply in traps is so labor intensive as to be likely unfeasible in crowded residential areas, and it is unclear how much more effective active traps lacking attractants are than purely passive traps. In this study, we analyzed Aedes capture data collected in 2019 from six urban areas in Kaohsiung City to compare Aedes mosquito catch rates between (passive) gravitraps and (active) fan-traps. The average gravitrap index (GI) and fan-trap index (FI) values were 0.68 and 3.39 respectively at peak catch times from June to August 2019, with consistently higher FI values calculated in all areas studied. We compared trap indices to reported cases of dengue fever and correlated them with weekly fluctuations in temperature and rainfall. We found that FI trends aligned more closely with case numbers and rainfall than GI values, supporting the use of fan-traps for Aedes mosquito surveillance and control as part of broader vector management strategies. Furthermore, combining fan-trap catch data with rapid testing for dengue infections may improve the early identification and prevention of future disease outbreaks.

Use of seroprevalence to guide dengue vaccination plans for older adults in a dengue non-endemic country.

A shift in dengue cases toward the adult population, accompanied by an increased risk of severe cases of dengue in the elderly, has created an important emerging issue in the past decade. To understand the level of past DENV infection among older adults after a large dengue outbreak occurred in southern Taiwan in 2015, we screened 1498 and 2603 serum samples from healthy residents aged ≥ 40 years in Kaohsiung City and Tainan City, respectively, to assess the seroprevalence of anti-DENV IgG in 2016. Seropositive samples were verified to exclude cross-reaction from Japanese encephalitis virus (JEV), using DENV/JEV-NS1 indirect IgG ELISA. We further identified viral serotypes and secondary DENV infections among positive samples in the two cities. The overall age-standardized seroprevalence of DENV-IgG among participants was 25.77% in Kaohsiung and 11.40% in Tainan, and the seroprevalence was significantly higher in older age groups of both cities. Although the percentages of secondary DENV infection in Kaohsiung and Tainan were very similar (43.09% and 44.76%, respectively), DENV-1 and DENV-2 spanned a wider age range in Kaohsiung, whereas DENV-2 was dominant in Tainan. As very few studies have obtained the serostatus of DENV infection in older adults and the elderly, this study highlights the need for further investigation into antibody status, as well as the safety and efficacy of dengue vaccination in these older populations.

Releasing Intracellular NS1 from Mosquito Cells for the Detection of Dengue Virus-Infected Mosquitoes.

Dengue virus (DENV), the pathogen that causes dengue fever, is mainly transmitted by Aedes aegypti. Surveillance of infected mosquitoes is a major component of integrated mosquito control methods for reducing the risk of vector-born disease outbreaks. However, a specialized rapid test for DENV detection in mosquitoes is not currently available. Utilizing immunoblotting, we found that the secretion of NS1 from both a DENV-infected mosquito cell line and mosquito bodies was below the detection threshold. However, when Triton X-100 was used to lyse infected mosquitoes, intracellular NS1 was released, and could then be effectively detected by the NS1 rapid test. The distribution of DENV NS1 in intrathoracically infected mosquitoes was different from that of orally infected mosquitoes. Next, we performed sensitivity tests by bisecting mosquitoes longitudinally; one half of each mosquito was subjected to the NS1 rapid test while the other half was used for qPCR confirmation. This modified test had a sensitivity of nearly 90% from five days post-infection onwards, while DENV had escaped from the midgut barrier. This adapted test offers a valuable, easy-to-use tool for mosquito surveillance, which is a crucial component of DENV disease control.

Epidemiological analysis of the Kaohsiung city strategy for dengue fever quarantine and epidemic prevention.

BACKGROUND: Dengue is endemic in over 100 countries and is an important public health problem worldwide. Dengue fever is not endemic in Taiwan; the importation of dengue viruses from neighboring countries via close commercial links and air travel is considered to be the cause of local outbreaks. Therefore, efforts toward disease control have focused on preventing the importation of dengue into Taiwan. In this study, we investigated the relationships between the numbers of imported and indigenous dengue cases to test the validity of this strategy. METHODS: Data on cases of dengue fever that occurred between 2013 and 2018 were obtained from the surveillance systems of the Taiwan Center for Disease Control and Kaohsiung City Health Department. Standard epidemiological data, including the monthly numbers of indigenous and imported cases of dengue, were calculated. Potential associations between the numbers of indigenous and imported cases were investigated using correlation analyses. RESULTS: We identified a possible relationship between the period of disease concealment and the number of imported dengue cases, which resulted in epidemics of indigenous dengue fever within local communities. Further analysis of confirmed cases during previous epidemics in Kaohsiung City found that the risk of indigenous dengue fever may be related to the likelihood that patients with imported dengue fever will stay within local communities. CONCLUSION: Given the correlations found between imported and indigenous cases of dengue fever, as well as the relationship between the disease concealment period and the risk of indigenous dengue fever, prevention of disease importation and efficient identification of dengue cases within high-risk communities remain the major priorities for disease control.

Dengue fever scoring system: new strategy for the early detection of acute dengue virus infection in Taiwan.

BACKGROUND/PURPOSE: Dengue fever is an important public health problem in Southern Taiwan. The purpose of this study was to develop a dengue scoring system using a three-stage process, which may be used as a guidance tool for the early diagnosis of dengue fever. METHODS: A retrospective study was conducted to identify factors useful for the early diagnosis of dengue fever. We assessed the clinical and laboratory features of 89 adult patients with dengue from 2002 to 2004 at a community-based hospital. They were compared with 14 patients with scrub typhus, 104 with Q fever, and 35 with murine typhus, which might present similar symptoms and signs as dengue infection. A scoring system was designed after analysis of the retrospective study and with the assistance of 10 expert clinicians. For the second stage, we evaluated efficiency in differentiating dengue fever from Q fever, scrub typhus and murine typhus in three hospitals from 2002 to 2005. For the third stage, we prospectively used the dengue scoring system for 498 cases that clinically were suspected as having dengue infection in the city of Kaohsiung from January 2006 to September 2006. RESULTS: The performance of the scoring system was 88.1% sensitivity, 94.9% specificity, 95.7% positive predictive value (PPV), and 86.1% negative predictive value (NPV). Evaluation of the scoring system at the third stage revealed 90.7% sensitivity, 86.9% specificity, 81.4% PPV, and 93.6% NPV. CONCLUSION: The dengue scoring system had a high NPV that might be helpful in the early diagnosis of dengue fever in adults before laboratory data are available.