Utilizing citizen science to model the distribution of Aedes aegypti in West Africa

In a rapidly urbanizing region such as West Africa, Aedes mosquitoes pose an emerging threat of infectious disease that is compounded by limited vector surveillance. Citizen science has been proposed as a way to fill surveillance gaps by training local residents to collect and share information on disease vectors. Increasing citizen science efforts can begin to bridge the gaps in our current knowledge of Aedes distribution while engaging locals with mosquito control and public health efforts. Understanding the distribution of disease vectors in West Africa can inform researchers and public health officials on where to conduct disease surveillance and focus public health interventions. We aimed to compare citizen science data to published literature observations of Aedes mosquitoes and to quantify how incorporating citizen science changes our understanding of Aedes mosquito distribution in West Africa. We utilized citizen science data collected through NASAs GLOBE Observer mobile phone application and data from a previously published literature review on Aedes mosquito distribution to examine the contribution of citizen science to understanding the distribution of Ae. aegypti in West Africa using Maximum Entropy modeling. Combining citizen science and literature-derived observations improved the fit of the model compared to models created by each data source alone, but did not alleviate location bias within the models, likely due to lack of widespread observations. Understanding Ae. aegypti distribution will require greater investment in Aedes mosquito surveillance in the region, and citizen science should be utilized as a tool in this mission to increase the reach of surveillance.

Mapping environmental suitability of Haemagogus and Sabethes spp. mosquitoes to understand sylvatic transmission risk of yellow fever virus in Brazil

Background Yellow fever (YF) is an arboviral disease which is endemic to Brazil due to a sylvatic transmission cycle maintained by infected mosquito vectors, non-human primate (NHP) hosts, and humans. Despite the existence of an effective vaccine, recent sporadic YF epidemics have underscored concerns about sylvatic vector surveillance, as very little is known about their spatial distribution. Here, we model and map the environmental suitability of YF’s main vectors in Brazil, Haemagogus spp. and Sabethes spp., and use human population and NHP data to identify locations prone to transmission and spillover risk. Methodology/Principal findings We compiled a comprehensive set of occurrence records on Hg. janthinomys, Hg. leucocelaenus, and Sabethes spp. from 1991–2019 using primary and secondary data sources. Linking these data with selected environmental and land-cover variables, we adopted a stacked regression ensemble modelling approach (elastic-net regularized GLM, extreme gradient boosted regression trees, and random forest) to predict the environmental suitability of these species across Brazil at a 1x1 km resolution. We show that while suitability for each species varies spatially, high suitability for all species was predicted in the Southeastern region where recent outbreaks have occurred. By integrating data on NHP host reservoirs and human populations, our risk maps further highlight municipalities within the region that are prone to transmission and spillover. Conclusions/Significance Our maps of sylvatic vector suitability can help elucidate potential locations of sylvatic reservoirs and be used as a tool to help mitigate risk of future YF outbreaks and assist in vector surveillance. Furthermore, at-risk regions identified from our work could help disease control and elucidate gaps in vaccination coverage and NHP host surveillance.

Aedes aegypti abundance, larval indices and risk for dengue virus transmission in Kinondoni district, Tanzania

Background Tanzania has experienced periodic dengue outbreaks with increased incidence since 2010. However, there is limited information on vector dynamics and transmission risk in most parts of the country. This study was conducted to determine Aedes mosquito abundance, larval indices and dengue virus infection rate as risk indicators for DENV transmission in Kinondoni district, Dar es Salaam, Tanzania. Methods A cross-sectional study was conducted in three wards of Kinondoni district in Tanzania between December 2019 and January 2020. In each ward, three streets were randomly selected for adult and immature mosquito sampling. The adult mosquitoes were collected using Mosquito Magnet traps, while mosquito larvae and pupae were inspected in water-holding containers in the selected household compounds. The detection of dengue virus (DENV) in female Aedes mosquitoes was done using a one-step reverse transcription–polymerase chain reaction (RT–PCR) method. Results Of the 1416 adult female mosquitoes collected, Ae. aegypti accounted for 16.8% (n = 238). A total of 333 water-holding containers were inspected and 201 (60.4%) had at least an Aedes larvae or pupae. Water-holding containers supporting the breeding of Aedes larvae and pupae included discarded car tires, flowerpots and small and large plastic containers. The overall House Index, Container Index and Breteau Index were 55.1%, 60.4% and 114.2, respectively. None of the 763 female Aedes mosquitoes tested by RT–PCR was found to be infected with DENV. Conclusion The presence and abundance Ae. aegypti mosquitoes and the large proportion of water-holding containers infested with the mosquito larvae and pupae put residents of Kinondoni district at high risk of DENV transmission. Our findings emphasize the need for continuous mosquito vector surveillance and control to prevent the possibility of future DENV outbreaks in Tanzania.

EFFECTIVENESS OF COMPREHENSIVE CAMPAIGN FOR DENGUE CONTROL IN SOUTHERN PUNJAB, PAKISTAN - AN EXPERIENCE OF 2 YEARS

Objective: To determine the effect of comprehensive campaign for dengue control in Southern Punjab, Pakistan. Study Design: Cross sectional study. Place and Duration of the Study: Station Health Organization Multan and Departments of Pathology, Combined Military Hospital Multan, from Mar 2019 to Dec 2020. Methodology: This study was conducted by implementing a comprehensive campaign by involvement of public health workers in high risk dengue area of Multan. There were five phases of this campaign; awareness, situation analysis, vector surveillance, indoor residual sprays where vector was found and contact tracing of dengue fever cases. The system was developed in 46 Mohallas/localities of Multan. Hot spot for mosquitoes breeding were identified at 15 places. Total of 23 health workers were recruited including 3 female workers. Insecticide spraying was carried out by indoor residual spray pumps & thermal foggers in a cyclic pattern. Results: During 2019, 42 larvae were identified followed by insecticide sprays & follow up visits; 26 cases of dengue fever were diagnosed and treated at CMH Multan in a dedicated dengue ward. Contact tracing was carried out and indoor residual sprays were carried out in 20 houses around residence of these cases. In 2020, 5 larvae were identified followed by sprays & follow up visits; 2 cases of dengue fever were diagnosed and treated. Conclusion: Comprehensive campaign against dengue fever was quite successful to control spread of dengue fever.

Rapid Differentiation of Vector Species: Development of Microsatellite Markers for Population Genetics of Biting Midges and a Potential Tool for Species Identification of Culicoides Sonorensis

Background: Proper vector surveillance relies on the ability to identify species of interest accurately and efficiently, though this can be difficult in groups containing cryptic species. Culicoides is a genus of small biting flies responsible for the transmission of numerous pathogens to a multitude of vertebrates. Regarding pathogen transmission, the C. variipennis species complex is of particular interest in North America. Of the six species within this group, only C. sonorensis is a proven vector of bluetongue virus and epizootic hemorrhagic disease virus. Unfortunately, subtle morphological differences, cryptic species, and mitonuclear discordance make species identification in the C. variipennis complex challenging. Recently, a SNP analysis enabled discrimination between the species of this group; however, this demanding approach is not practical for vector surveillance. Methods: The aim of the current study was to develop a reliable and affordable way of differentiating the species within the C. variipennis complex, especially C. sonorensis. Twenty-five putative microsatellite markers were identified using the C. sonorensis genome and tested for amplification within five species of the C. variipennis complex. Machine learning was then used to determine which markers best explain the genetic differentiation between species. This led to the development of a subset of four and seven markers which were also tested for species differentiation.Results: A total of 21 microsatellite markers were successfully amplified in the species tested. Clustering analyses of all of these markers recover the same species-level identification as the previous SNP data. Additionally, the subset of seven markers was equally capable of accurately differentiating the members of the C. variipennis complex as the 21 microsatellite markers. Finally, one microsatellite marker (C508) was found to be species-specific, only amplifying in the vector species C. sonorensis among the samples tested. Conclusions: These microsatellites provide an affordable way in which to differentiate the species of the C. variipennis complex and could lead to a better understanding of the species dynamics within this group. Additionally, after further testing, marker C508 may allow for the identification of C. sonorensis with a single-tube assay, potentially providing a powerful new tool for vector surveillance in North America.

Anopheline Diversity in Indonesia: An Evaluation of Animal-Baited Sampling Techniques

Indonesia has rich Anopheline (Diptera: Culicidae) mosquito species living in various types of ecosystems. The study was conducted to profile and compare Anopheles diversity, equitability, and dominance in various ecosystems using different animal-based sampling techniques. The present study analyzed a subset of data collected from a nation-wide vector and animal reservoirs survey in 2016. Analyses were restricted to three ecosystem types (forest, nonforest, and coastal areas) in Java and Sumatera Islands. A total of 5,477 Anopheles were collected by using animal-baited (n = 1,909) and animal-baited trap nets (n = 1,978), consisting of 23 Anopheline species. Overall, Anopheles vagus was the most abundant species, followed by An. subpictus and An. barbirostris. Among the three ecosystems, the forest had a higher diversity index (H′ = 1.98), but each ecosystem has its specific predominant species. Compared with the animal-baited method, the Anopheles abundance collected by animal-baited trap nets was two-fold higher. Ecosystem, elevation, and sampling methods were associated with the abundance of female Anopheles (P-value < 0.001). Our findings revealed that Anopheles were found in a different ecosystem, indicating the potential of malaria transmission. This suggests that improved malaria vector surveillance is essential in all types of ecosystem. Furthermore, the study suggested that animal-baited trap nets could be used as the standard method of outdoor resting sampling in Indonesia in addition to the traditional human landing collection approach.

Effectiveness of joint 3 + 1 malaria strategy along China–Myanmar cross border areas

Background Cross-border malaria in Laiza City of Myanmar seriously affected Yingjiang County of China and compromised reaching the goal of malaria elimination by 2020. Since 2017, a pilot project on 3 + 1 strategy of joint cross-border malaria prevention and control was carried out for building a malaria buffer in these border areas. Here, 3 were the three preventive lines in China where different focalized approaches of malaria elimination were applied and + 1 was a defined border area in Myanmar where the integrated measures of malaria control were adopted. Methods A 5-year retrospective analysis (2015 to 2019) was conducted that included case detection, parasite prevalence and vector surveillance. Descriptive statistics was used and the incidence or rates were compared. The annual parasite incidence and the parasite prevalence rate in + 1 area of Myanmar, the annual importation rate in Yingjiang County of China and the density of An. minimus were statistically significant indictors to assess the effectiveness of the 3 + 1 strategy. Results In + 1 area of Myanmar from 2015 to 2019, the averaged annual parasite incidence was (59.11 ± 40.73)/1000 and Plasmodium vivax accounted for 96.27% of the total confirmed cases. After the pilot project, the annual parasite incidence dropped 89% from 104.77/1000 in 2016 to 12.18/1000 in 2019, the microscopic parasite prevalence rate dropped 100% from 0.34% in 2017 to zero in 2019 and the averaged density of An. Minimus per trap-night dropped 93% from 1.92 in June to 0.13 in September. The submicroscopic parasite prevalence rate increased from 1.15% in 2017 to 1.66% in 2019 without significant difference between the two surveys (P = 0.084). In Yingjiang County of China, neither indigenous nor introduced case was reported and 100% cases were imported from Myanmar since 2017. The averaged annual importation rate from 2015 to 2019 was (0.47 ± 0.15)/1000. After the pilot project, the annual importation rate dropped from 0.59/1000 in 2016 to 0.28/1000 in 2019 with an overall reduction of 53% in the whole county. The reduction was 67% (57.63/1000 to 18.01/1000) in the first preventive line, 52% (0.20/1000 to 0.10/1000) in the second preventive line and 36% (0.32/1000 to 0.22/1000) in the third preventive line. The averaged density of An. Minimus per trap-night in the first preventive line dropped 94% from 2.55 in June to 0.14 in September, without significant difference from that of + 1 area of Myanmar (Z value = − 1.18, P value = 0.24). Conclusion The pilot project on 3 + 1 strategy has been significantly effective in the study areas and a buffer zone of border malaria was successfully established between Laiza City of Myanmar and Yingjiang County of China.

Comparing Light—Emitting—Diodes Light Traps for Catching Anopheles Mosquitoes in a Forest Setting, Western Thailand

Light traps are a common method for attracting and collecting arthropods, including disease vectors such as mosquitoes. Various types of traps have been used to monitor mosquitoes in a forest in Western Thailand. In this study, four Light Emitting Diodes (LED) light sources (UV, blue, green, and red) and two fluorescent lights (white and UV) were used to trap nocturnal adult mosquitoes. These traps were used with light alone and not any additional attractant. The experiment was conducted from 18:00 to 06:00 h. on six consecutive nights, every two months, across dry, wet, and cold seasons. All specimens were first identified by morphological features and subsequently confirmed by using PCR. We collected a total of 873 specimens of 31 species in four genera, Anopheles, Aedes, Culex, and Armigeres. Anopheles harrisoni was the predominant species, followed by Aedes albopictus, Culex brevipalpis, Culex nitropunctatus, and Armigeres (Leicesteria) longipalpis. UV fluorescent light was the most effective light source for capturing forest mosquitoes, followed by UV LED, blue LED, green LED, white fluorescent, and red LED. The optimal times for collection were from 21:00 to 03:00 h in the dry season. Our results demonstrate that appropriate sampling times and light sources should be selected for optimal efficiency in vector surveillance programs.

Laboratory and Field Evaluations of a Commercially Available Real-Time Loop-Mediated Isothermal Amplification Assay for the Detection of West Nile Virus in Mosquito Pools

ABSTRACT Although the specific cDNA amplification mechanisms of reverse-transcriptase polymerase chain reaction (RT-PCR) and RT loop-mediated isothermal amplification (RT-LAMP) are very different, both molecular assays serve as options to detect arboviral RNA in mosquito pools. Like RT-PCR, RT-LAMP uses a reverse transcription step to synthesize complementary DNA (cDNA) from an RNA template and then uses target-specific primers to amplify cDNA to detectable levels in a single-tube reaction. Using laboratory-generated West Nile virus (WNV) samples and field-collected mosquito pools, we evaluated the sensitivity and specificity of a commercially available WNV real-time RT-LAMP assay (Pro-AmpRT™ WNV; Pro-Lab Diagnostics, Inc., Round Rock, Texas) and compared the results to a validated real-time RT-PCR assay. Laboratory generated virus stock samples containing ≥ 2.3 log10 plaque-forming units (PFU)/ml and intrathoracically inoculated mosquitoes containing ≥ 2.4 log10 PFU/ml produced positive results in the Pro-AmpRT WNV assay. Of field-collected pools that were WNV positive by real-time RT-PCR, 74.5% (70 of 94) were also positive by the Pro-AmpRT WNV assay, resulting in an overall Cohen's kappa agreement of 79.4% between the 2 tests. The Pro-AmpRT WNV assay shows promise as a suitable virus screening tool for vector surveillance programs provided agencies are aware of its characteristics and limitations.

Knowledge discovery and dengue forecasting applied in a four-year dataset collected at Natal, RN – Brazil

Dengue, a disease recognized as a health problem, causes significant impacts on health and affects millions of people each year worldwide. A suitable method for dengue vector surveillance is to count eggs the mosquitoes Aedes aegypti have laid in spatially distributed ovitraps. In view of this approach, this study uses a database collected in 397 ovitraps distributed across the municipality of Natal, RN – Brazil. The number of eggs in each ovitrap was counted weekly, over four years (2016 - 2019), and simultaneously analyzed with the incidence of dengue. Our results confirm that dengue incidence seems to be related to socioeconomic status in Natal. Using a deep learning model, we then predict the incidence of new dengue cases based on data obtained from the previous week of dengue or the number of eggs present in the ovitraps. The analysis shows that ovitrap data allows earlier detection (four to six weeks) when compared to dengue cases themselves (one week). Furthermore, the results confirm that quantifying Aedes aegypti eggs may be valuable for planning actions and public health interventions.