Genomic profiling of climate adaptation in Aedes aegypti along an altitudinal gradient in Nepal indicates nongradual expansion of the disease vector.

Driven by globalization, urbanization and climate change, the distribution range of invasive vector species has expanded to previously colder ecoregions. To reduce health-threatening impacts on humans, insect vectors are extensively studied. Population genomics can reveal the genomic basis of adaptation and help to identify emerging trends of vector expansion. By applying whole genome analyses and genotype-environment associations to populations of the main dengue vector Aedes aegypti, sampled along an altitudinal gradient in Nepal (200-1300 m), we identify putatively adaptive traits and describe the species' genomic footprint of climate adaptation to colder ecoregions. We found two differentiated clusters with significantly different allele frequencies in genes associated to climate adaptation between the highland population (1300 m) and all other lowland populations (≤800 m). We revealed nonsynonymous mutations in 13 of the candidate genes associated to either altitude, precipitation or cold tolerance and identified an isolation-by-environment differentiation pattern. Other than the expected gradual differentiation along the altitudinal gradient, our results reveal a distinct genomic differentiation of the highland population. Local high-altitude adaptation could be one explanation of the population's phenotypic cold tolerance. Carrying alleles relevant for survival under colder climate increases the likelihood of this highland population to a worldwide expansion into other colder ecoregions.

The knowledge, attitude and practice of community people on dengue fever in Central Nepal: a cross-sectional study.

BACKGROUND: Since 2006, Nepal has experienced frequent Dengue fever (DF) outbreaks. Up to now, there have been no knowledge, attitude and practice (KAP) studies carried out on DF in Nepal that have included qualitative in-depth and quantitative data. Thus, we aimed to explore and compare the KAP of people residing in the lowland (< 1500 m) and highland (> 1500 m) areas of Nepal. METHODS: A cross-sectional mixed-method study was conducted in six districts of central Nepal in September-October 2018 including both quantitative (660 household surveys) and qualitative data (12 focus group discussions and 27 in-depth interviews). The KAP assessment was executed using a scoring system and defined as high or low based on 80% cut-off point. Logistic regression was used to investigate the associated factors, in quantitative analysis. The deductive followed by inductive approach was adopted to identify the themes in the qualitative data. RESULTS: The study revealed that both the awareness about DF and prevention measures were low. Among the surveyed participants, 40.6% had previously heard about DF with a significantly higher number in the lowland areas. Similarly, IDI and FGD participants from the lowland areas were aware about DF, and it's associated symptoms, hence they were adopting better preventive practices against DF. The findings of both the qualitative and quantitative data indicate that people residing in the lowland areas had better knowledge on DF compared to people in highland areas. All IDI participants perceived a higher chance of increasing future dengue outbreaks due to increasing temperature and the mobility of infected people from endemic to non-endemic areas. The most quoted sources of information were the television (71.8%) and radio (51.5%). Overall, only 2.3% of the HHS participants obtained high knowledge scores, 74.1% obtained high attitude scores and 21.2% obtained high preventive practice scores on DF. Among the socio-demographic variables, the area of residence, educational level, age, monthly income, SES and occupation were independent predictors of knowledge level, while the education level of the participants was an independent predictor of the attitude level. CONCLUSIONS: Our study found a very low level of knowledge and insufficient preventive practices. This highlights an urgent need for extensive dengue prevention programs in both highland and lowland communities of Nepal.

Phenotypic insecticide resistance status of the Culex pipiens complex: a European perspective.

BACKGROUND: The common house mosquito Culex pipiens is known to be a major vector for West Nile virus. In order to decrease risks of West Nile virus outbreaks in Europe, insecticides and the bio-larvicide Bacillus thuringiensis israelensis (Bti) are commonly used for vector control. Alarmingly, insecticide resistance has been reported in Cx. pipiens populations from Southern Europe and several countries neighbouring Europe. For Central and Northern Europe, however, the phenotypic insecticide resistance status of Cx. pipiens has not yet been investigated. METHODS: A literature review was performed to assess the geographical distribution of insecticide resistance in Cx. pipiens. To fill the gap of knowledge for Central and Northern Europe, WHO susceptibility tests with permethrin, deltamethrin, malathion, bendiocarb and DDT and a larval toxicity test with Bti were performed with a Cx. pipiens population from Belgium, a country in Central Europe. RESULTS: This research provides the first evidence of widespread phenotypic insecticide resistance in Cx. pipiens. In general, Cx. pipiens developed resistance against multiple insecticides in several countries. Another Cx. pipiens population from Belgium was tested and showed insecticide resistance against deltamethrin, permethrin, DDT and possibly against bendiocarb. The bio-larvicide Bti caused lower mortality than reported for other Cx. pipiens populations in the literature. CONCLUSIONS: These results indicate the urgent need for insecticide resistance monitoring against commonly used adulticides and larvicides in Europe, for the translation of knowledge gained regarding the limited efficiency and availability of insecticide into EU legislation and the need for innovative non-chemical vector control tools in order to counter the widespread insecticide resistance in Culex populations.

A barcoding pipeline for mosquito surveillance in Nepal, a biodiverse dengue-endemic country.

BACKGROUND: Vector-borne diseases are on the rise on a global scale, which is anticipated to further accelerate because of anthropogenic climate change. Resource-limited regions are especially hard hit by this increment with the currently implemented surveillance programs being inadequate for the observed expansion of potential vector species. Cost-effective methods that can be easily implemented in resource-limited settings, e.g. under field conditions, are thus urgently needed to function as an early warning system for vector-borne disease epidemics. Our aim was to enhance entomological capacity in Nepal, a country with endemicity of numerous vector-borne diseases and with frequent outbreaks of dengue fever. METHODS: We used a field barcoding pipeline based on DNA nanopore sequencing (Oxford Nanopore Technologies) and verified its use for different mosquito life stages and storage methods. We furthermore hosted an online workshop to facilitate knowledge transfer to Nepalese scientific experts from different disciplines. RESULTS: The use of the barcoding pipeline could be verified for adult mosquitos and eggs, as well as for homogenized samples, dried specimens, samples that were stored in ethanol and frozen tissue. The transfer of knowledge was successful, as reflected by feedback from the participants and their wish to implement the method. CONCLUSIONS: Cost effective strategies are urgently needed to assess the likelihood of disease outbreaks. We were able to show that field sequencing provides a solution that is cost-effective, undemanding in its implementation and easy to learn. The knowledge transfer to Nepalese scientific experts from different disciplines provides an opportunity for sustainable implementation of low-cost portable sequencing solutions in Nepal.

The ecophysiological plasticity of Aedes aegypti and Aedes albopictus concerning overwintering in cooler ecoregions is driven by local climate and acclimation capacity.

Aedes aegypti and Aedes albopictus transmit diseases such as dengue, and are of major public health concern. Driven by climate change and global trade/travel both species have recently spread to new tropic/subtropic regions and Ae. albopictus also to temperate ecoregions. The capacity of both species to adapt to new environments depends on their ecophysiological plasticity, which is the width of functional niches where a species can survive. Mechanistic distribution models often neglect to incorporate ecophysiological plasticity especially in regards to overwintering capacity in cooler habitats. To portray the ecophysiological plasticity concerning overwintering capability, we conducted temperature experiments with multiple populations of both species originating from an altitudinal gradient in South Asia and tested as follows: the cold tolerance of eggs (-2 °C- 8 days and - 6 °C- 2 days) without and with an experimental winter onset (acclimation: 10 °C- 60 days), differences between a South Asian and a European Ae. albopictus population and the temperature response in life cycles (13 °C, 18 °C, 23 °C, 28 °C). Ecophysiological plasticity in overwintering capacity in Ae. aegypti is high in populations originating from low altitude and in Ae. albopictus populations from high altitude. Overall, ecophysiological plasticity is higher in Ae. albopictus compared to Ae. aegypti. In both species acclimation and in Ae. albopictus temperate continental origin had a huge positive effect on survival. Our results indicate that future mechanistic prediction models can include data on winter survivorship of both, tropic and subtropic Ae. aegypti, whereas for Ae. albopictus this depends on the respective temperate, tropical region the model is focusing on. Future research should address cold tolerance in multiple populations worldwide to evaluate the full potential of the ecophysiological plasticity in the two species. Furthermore, we found that Ae. aegypti can survive winter cold especially when acclimated and will probably further spread to colder ecoregions driven by climate change.

Spatiotemporal Distribution of Dengue and Chikungunya in the Hindu Kush Himalayan Region: A Systematic Review.

The risk of increasing dengue (DEN) and chikungunya (CHIK) epidemics impacts 240 million people, health systems, and the economy in the Hindu Kush Himalayan (HKH) region. The aim of this systematic review is to monitor trends in the distribution and spread of DEN/CHIK over time and geographically for future reliable vector and disease control in the HKH region. We conducted a systematic review of the literature on the spatiotemporal distribution of DEN/CHIK in HKH published up to 23 January 2020, following Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. In total, we found 61 articles that focused on the spatial and temporal distribution of 72,715 DEN and 2334 CHIK cases in the HKH region from 1951 to 2020. DEN incidence occurs in seven HKH countries, i.e., India, Nepal, Bhutan, Pakistan, Bangladesh, Afghanistan, and Myanmar, and CHIK occurs in four HKH countries, i.e., India, Nepal, Bhutan, and Myanmar, out of eight HKH countries. DEN is highly seasonal and starts with the onset of the monsoon (July in India and June in Nepal) and with the onset of spring (May in Bhutan) and peaks in the postmonsoon season (September to November). This current trend of increasing numbers of both diseases in many countries of the HKH region requires coordination of response efforts to prevent and control the future expansion of those vector-borne diseases to nonendemic areas, across national borders.