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Dengue, Zika, chikungunya and yellow fever viruses continue to be a major public health burden. Aedes mosquitoes, the primary vectors responsible for transmitting these viral pathogens, continue to flourish due to local challenges in vector control management. Yeast interfering RNA-baited larval lethal ovitraps are being developed as a novel biorational control tool for Aedes mosquitoes. This intervention circumvents increasing issues with insecticide resistance and poses no known threat to non-target organisms. In an effort to create public awareness of this alternative vector control strategy, gain stakeholder feedback regarding product design and acceptance of the new intervention, and build capacity for its potential integration into existing mosquito control programs, this investigation pursued community stakeholder engagement activities, which were undertaken in Trinidad and Tobago. Three forms of assessment, including paper surveys, community forums, and household interviews, were used with the goal of evaluating local community stakeholders' knowledge of mosquitoes, vector control practices, and perceptions of the new technology. These activities facilitated evaluation of the hypothesis that the ovitraps would be broadly accepted by community stakeholders as a means of biorational control for Aedes mosquitoes. A comparison of the types of stakeholder input communicated through use of the three assessment tools highlighted the utility and merit of using each tool for assessing new global health interventions. Most study participants reported a general willingness to purchase an ovitrap on condition that it would be affordable and safe for human health and the environment. Stakeholders provided valuable input on product design, distribution, and operation. A need for educational campaigns that provide a mechanism for educating stakeholders about vector ecology and management was highlighted. The results of the investigation, which are likely applicable to many other Caribbean nations and other countries with heavy arboviral disease burdens, were supportive of supplementation of existing vector control strategies through the use of the yeast RNAi-based ovitraps., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Scheel and Severson are inventors on a related patent submission, US patent application 62/361,704 / European patent application EP 17828458.4. This does not alter their adherence to PLOS ONE policies on sharing data and materials. The other authors have no competing interests.

Mosquito vectors in the genera Anopheles, Aedes, and Culex transmit a variety of medically important pathogens. Current vector control tools are reaching the limits of their effectiveness, necessitating the introduction of innovative vector control technologies. RNAi, which facilitates functional characterization of mosquito genes in the laboratory, could one day be applied as a new method of vector control. Recent advances in the oral administration of microbial-based systems for delivery of species-specific interfering RNA pesticides to mosquitoes may facilitate translation of this technology to the field. Oral RNAi-based pesticides represent a new class of biorational pesticides that could combat increased global incidence of insecticide resistance and which could one day become critical components of integrated human disease vector mosquito control programs., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Documenting isolation is notoriously difficult for species with vast polymorphic populations. High proportions of shared variation impede estimation of connectivity, even despite leveraging information from many genetic markers. We overcome these impediments by combining classical analysis of neutral variation with assays of the structure of selected variation, demonstrated using populations of the principal African malaria vector Anopheles gambiae . Accurate estimation of mosquito migration is crucial for efforts to combat malaria. Modeling and cage experiments suggest that mosquito gene drive systems will enable malaria eradication, but establishing safety and efficacy requires identification of isolated populations in which to conduct field testing. We assess Lake Victoria islands as candidate sites, finding one island 30 km offshore is as differentiated from mainland samples as populations from across the continent. Collectively, our results suggest sufficient contemporary isolation of these islands to warrant consideration as field-testing locations and illustrate shared adaptive variation as a useful proxy for connectivity in highly polymorphic species., Competing Interests: The authors declare no competing financial interests., (© 2019 The Authors. Evolutionary Applications published by John Wiley & Sons Ltd.)

Background: Malaria is the leading cause of global paediatric mortality in children below 5 years of age. The number of fatalities has reduced significantly due to an expansion of control interventions but the development of new technologies remains necessary in order to achieve elimination. Recent attention has been focused on the release of genetically modified (GM) mosquitoes into natural vector populations as a mechanism of interrupting parasite transmission but despite successful in vivo laboratory studies, a detailed population genetic assessment, which must first precede any proposed field trial, has yet to be undertaken systematically. Here, the genetic structure of Anopheles gambiae populations in north-western Lake Victoria is explored to assess their suitability as candidates for a pilot field study release of GM mosquitoes., Methods: 478 Anopheles gambiae mosquitoes were collected from six locations and a subset (N = 96) was selected for restriction site-associated DNA sequencing (RADseq). The resulting single nucleotide polymorphism (SNP) marker set was analysed for effective size (N e ), connectivity and population structure (PCA, F ST )., Results: 5175 high-quality genome-wide SNPs were identified. A principal components analysis (PCA) of the collinear genomic regions illustrated that individuals clustered in concordance with geographic origin with some overlap between sites. Genetic differentiation between populations was varied with inter-island comparisons having the highest values (median F ST 0.0480-0.0846). N e estimates were generally small (124.2-1920.3)., Conclusions: A reduced-representation SNP marker set for genome-wide An. gambiae genetic analysis in the north-western Lake Victoria basin is reported. Island populations demonstrated low to moderate genetic differentiation and greater structure suggesting some limitation to migration. Smaller estimates of N e indicate that an introduced effector transgene will be more susceptible to genetic drift but to ensure that it is driven to fixation a robust gene drive mechanism will likely be needed. These findings, together with their favourable location and suitability for frequent monitoring, indicate that the Ssese Islands contain several candidate field locations, which merit further evaluation as potential GM mosquito pilot release sites.

Background: Understanding population genetic structure in the malaria vector Anopheles gambiae (s.s.) is crucial to inform genetic control and manage insecticide resistance. Unfortunately, species characteristics such as high nucleotide diversity, large effective population size, recent range expansion, and high dispersal ability complicate the inference of genetic structure across its range in sub-Saharan Africa. The ocean, along with the Great Rift Valley, is one of the few recognized barriers to gene flow in this species, but the effect of inland lakes, which could be useful sites for initial testing of genetic control strategies, is relatively understudied. Here we examine Lake Victoria as a barrier between the Ugandan mainland and the Ssese Islands, which lie up to 60 km offshore. We use mitochondrial DNA (mtDNA) from populations sampled in 2002, 2012 and 2015, and perform Bayesian cluster analysis on mtDNA combined with microsatellite data previously generated from the same 2002 mosquito DNA samples., Results: Hierarchical analysis of molecular variance and Bayesian clustering support significant differentiation between the mainland and lacustrine islands. In an mtDNA haplotype network constructed from this and previous data, haplotypes are shared even between localities separated by the Rift Valley, a result that more likely reflects retention of shared ancestral polymorphism than contemporary gene flow., Conclusions: The relative genetic isolation of An. gambiae on the Ssese Islands, their small size, level terrain and ease of access from the mainland, the relative simplicity of the vectorial system, and the prevalence of malaria, are all attributes that recommend these islands as possible sites for the testing of genetic control strategies.

Independent estimates of fossil fuel CO2 (CO2ff) emissions are key to ensuring that emission reductions and regulations are effective and provide needed transparency and trust. Point source emissions are a key target because a small number of power plants represent a large portion of total global emissions. Currently, emission rates are known only from self-reported data. Atmospheric observations have the potential to meet the need for independent evaluation, but useful results from this method have been elusive, due to challenges in distinguishing CO2ff emissions from the large and varying CO2 background and in relating atmospheric observations to emission flux rates with high accuracy. Here we use time-integrated observations of the radiocarbon content of CO2 ((14)CO2) to quantify the recently added CO2ff mole fraction at surface sites surrounding a point source. We demonstrate that both fast-growing plant material (grass) and CO2 collected by absorption into sodium hydroxide solution provide excellent time-integrated records of atmospheric (14)CO2 These time-integrated samples allow us to evaluate emissions over a period of days to weeks with only a modest number of measurements. Applying the same time integration in an atmospheric transport model eliminates the need to resolve highly variable short-term turbulence. Together these techniques allow us to independently evaluate point source CO2ff emission rates from atmospheric observations with uncertainties of better than 10%. This uncertainty represents an improvement by a factor of 2 over current bottom-up inventory estimates and previous atmospheric observation estimates and allows reliable independent evaluation of emissions., Competing Interests: The authors declare no conflict of interest.

Increasing reports of insecticide resistance continue to hamper the gains of vector control strategies in curbing malaria transmission. This makes identifying new insecticide targets or alternative vector control strategies necessary. CLassifier of Essentiality AcRoss EukaRyote (CLEARER), a leave-one-organism-out cross-validation machine learning classifier for essential genes, was used to predict essential genes in Anopheles gambiae and selected predicted genes experimentally validated. The CLEARER algorithm was trained on six model organisms: Caenorhabditis elegans, Drosophila melanogaster, Homo sapiens, Mus musculus, Saccharomyces cerevisiae and Schizosaccharomyces pombe, and employed to identify essential genes in An. gambiae. Of the 10,426 genes in An. gambiae, 1,946 genes (18.7%) were predicted to be Cellular Essential Genes (CEGs), 1716 (16.5%) to be Organism Essential Genes (OEGs), and 852 genes (8.2%) to be essential as both OEGs and CEGs. RNA interference (RNAi) was used to validate the top three highly expressed non-ribosomal predictions as probable vector control targets, by determining the effect of these genes on the survival of An. gambiae G3 mosquitoes. In addition, the effect of knockdown of arginase (AGAP008783) on Plasmodium berghei infection in mosquitoes was evaluated, an enzyme we computationally inferred earlier to be essential based on chokepoint analysis. Arginase and the top three genes, AGAP007406 (Elongation factor 1-alpha, Elf1), AGAP002076 (Heat shock 70kDa protein 1/8, HSP), AGAP009441 (Elongation factor 2, Elf2), had knockdown efficiencies of 91%, 75%, 63%, and 61%, respectively. While knockdown of HSP or Elf2 significantly reduced longevity of the mosquitoes (p<0.0001) compared to control groups, Elf1 or arginase knockdown had no effect on survival. However, arginase knockdown significantly reduced P. berghei oocytes counts in the midgut of mosquitoes when compared to LacZ-injected controls. The study reveals HSP and Elf2 as important contributors to mosquito survival and arginase as important for parasite development, hence placing them as possible targets for vector control. [ABSTRACT FROM AUTHOR]

Background: RNA interference (RNAi) is a target-specific gene silencing method that can be used to determine gene functions and investigate host–pathogen interactions, as well as facilitating the development of ecofriendly pesticides. Commercially available transfection reagents (TRs) can improve the efficacy of RNAi. However, we currently lack a product and protocol for the transfection of insect cell lines with long double-stranded RNA (dsRNA). Methods: We used agarose gel electrophoresis to determine the capacity of eight TRs to form complexes with long dsRNA. A CellTiter-Glo assay was then used to assess the cytotoxicity of the resulting lipoplexes. We also measured the cellular uptake of dsRNA by fluorescence microscopy using the fluorophore Cy3 as a label. Finally, we analyzed the TRs based on their transfection efficacy and compared the RNAi responses of Aedes albopictus C6/36 and U4.4 cells by knocking down an mCherry reporter Semliki Forest virus in both cell lines. Results: The TRs from Biontex (K4, Metafectene Pro, and Metafectene SI+) showed the best complexing capacity and the lowest dsRNA:TR ratio needed for complete complex formation. Only HiPerFect was unable to complex the dsRNA completely, even at a ratio of 1:9. Most of the complexes containing mCherry-dsRNA were nontoxic at 2 ng/µL, but Lipofectamine 2000 was toxic at 1 ng/µL in U4.4 cells and at 2 ng/µL in C6/36 cells. The transfection of U4.4 cells with mCherry-dsRNA/TR complexes achieved significant knockdown of the virus reporter. Comparison of the RNAi response in C6/36 and U4.4 cells suggested that C6/36 cells lack the antiviral RNAi response because there was no significant knockdown of the virus reporter in any of the treatments. Conclusions: C6/36 cells have an impaired RNAi response as previously reported. This investigation provides valuable information for future RNAi experiments by showing how to mitigate the adverse effects attributed to TRs. This will facilitate the judicious selection of TRs and transfection conditions conducive to RNAi research in mosquitoes. [ABSTRACT FROM AUTHOR]

Sustainable reductions in African malaria transmission require innovative tools for mosquito control. One proposal involves the use of low-threshold gene drive in Anopheles vector species, where a ‘causal pathway’ would be initiated by (i) the release of a gene drive system in target mosquito vector species, leading to (ii) its transmission to subsequent generations, (iii) its increase in frequency and spread in target mosquito populations, (iv) its simultaneous propagation of a linked genetic trait aimed at reducing vectorial capacity for Plasmodium, and (v) reduced vectorial capacity for parasites in target mosquito populations as the gene drive system reaches fixation in target mosquito populations, causing (vi) decreased malaria incidence and prevalence. Here the scope, objectives, trial design elements, and approaches to monitoring for initial field releases of such gene dive systems are considered, informed by the successful implementation of field trials of biological control agents, as well as other vector control tools, including insecticides, Wolbachia, larvicides, and attractive-toxic sugar bait systems. Specific research questions to be addressed in initial gene drive field trials are identified, and adaptive trial design is explored as a potentially constructive and flexible approach to facilitate testing of the causal pathway. A fundamental question for decision-makers for the first field trials will be whether there should be a selective focus on earlier points of the pathway, such as genetic efficacy via measurement of the increase in frequency and spread of the gene drive system in target populations, or on wider interrogation of the entire pathway including entomological and epidemiological efficacy. How and when epidemiological efficacy will eventually be assessed will be an essential consideration before decisions on any field trial protocols are finalized and implemented, regardless of whether initial field trials focus exclusively on the measurement of genetic efficacy, or on broader aspects of the causal pathway. Statistical and modelling tools are currently under active development and will inform such decisions on initial trial design, locations, and endpoints. Collectively, the considerations here advance the realization of developer ambitions for the first field trials of low-threshold gene drive for malaria vector control within the next 5 years. [ABSTRACT FROM AUTHOR]

Background: Sugar, when used as the phagostimulant in attractive toxic bait control tools, limits the efficacy and selectivity of this technology. Thus, more potent and selective phagostimulants than sugar are required to improve this technology. The potency of adenosine triphosphate (ATP) as an alternative model phagostimulant was assessed to determine its capacity to override the aversive effects of select antifeedants and toxicants. How ATP and sucrose modulate the rate of toxicity in the yellow fever mosquito Aedes aegypti was also examined. Methods: A no-choice feeding assay was used to investigate the phagostimulatory ability of ATP to override the aversive effects of structurally divergent antifeedant and toxicant compounds, and to modulate the rate of toxicity over 24 h. Binary combinations of antifeedant and toxicant compounds, at various concentrations, were similarly assessed for enhanced lethal potency. In comparison, no-choice open access and cotton wick feeding assays were used to determine the phagostimulatory role of sucrose in the ingestion of boric acid-laced diets. Dissections of the guts were performed to determine the diet destination as dependant on the phagostimulant. Results: ATP is a potent phagostimulant that dose dependently overrides aversion to antifeedant and toxicant tastants. Feeding on antifeedant- or toxicant-laced diets that was induced by ATP selectively resulted in rapid knockdown (nicotine, lobeline and caffeine) or death (boric acid and propylene glycol), with a combination of the two lethal compounds inducing a synergistic effect at lower concentrations. ATP- and sucrose-induced feeding predominantly directed the antifeedant- or toxicant-laced meals to the midgut and the crop, respectively. Conclusions: ATP is an efficacious alternative model phagostimulant to sucrose that overrides the aversive effects of antifeedants and toxicants, resulting in rapid toxic effects. Furthermore, this study demonstrates that variation in the rate of toxicity between ATP- and sugar-induced feeding is at least partly regulated by the differential feeding response, volume imbibed and the destination of the meals. Additional research is needed to identify structurally related, stable analogues of ATP due to the ephemeral nature of this molecule. For future applications, the workflow presented in this study may be used to evaluate such analogues for their suitability for use in attractive bait stations designed to target a broad range of haematophagous arthropods and prevent off-target species' feeding. [ABSTRACT FROM AUTHOR]

Background: Insecticide resistance in malaria vectors can be spatially highly heterogeneous, yet population structure analyses frequently find relatively high levels of gene flow among mosquito populations. Few studies have contemporaneously assessed phenotypic, genotypic and population structure analysis on mosquito populations and none at fine geographical scales. In this study, genetic diversity, population structure, and insecticide resistance profiles of Anopheles funestus and Anopheles arabiensis were examined across mosquito populations from and within neighbouring villages. Methods: Mosquitoes were collected from 11 towns in southern Mozambique, as well as from different neighbourhoods within the town of Palmeira, during the peak malaria transmission season in 2016. CDC bottle bioassay and PCR assays were performed with Anopheles mosquitoes at each site to determine phenotypic and molecular insecticide resistance profiles, respectively. Microsatellite analysis was conducted on a subsample of mosquitoes to estimate genetic diversity and population structure. Results: Phenotypic insecticide resistance to deltamethrin was observed in An. funestus sensu stricto (s.s.) throughout the area, though a high level of mortality variation was seen. However, 98% of An. funestus s.s. were CYP6P9a homozygous resistant. An. arabiensis was phenotypically susceptible to deltamethrin and 99% were kdr homozygous susceptible. Both Anopheles species exhibited high allelic richness and heterozygosity. Significant deviations from Hardy–Weinberg equilibrium were observed, and high linkage disequilibrium was seen for An. funestus s.s., supporting population subdivision. However, the FST values were low for both anophelines (− 0.00457 to 0.04213), Nm values were high (9.4–71.8 migrants per generation), AMOVA results showed almost 100% genetic variation among and within individuals, and Structure analysis showed no clustering of An. funestus s.s. and An. arabiensis populations. These results suggest high gene flow among mosquito populations. Conclusion: Despite a relatively high level of phenotypic variation in the An. funestus population, molecular analysis shows the population is admixed. These data indicate that CYP6P9a resistance markers do not capture all phenotypic variation in the area, but also that resistance genes of high impact are likely to easily spread in the area. Conversely, other strategies, such as transgenic mosquito release programmes will likely not face challenges in this locality. [ABSTRACT FROM AUTHOR]

Introduction: Mosquito-borne diseases have historically affected communities, especially in tropical areas where mosquitoes and illnesses are endemic. Globalization, climate change, and increased travel have created ideal conditions for outbreaks of mosquito-borne diseases that could threaten the American health system and place a burden on the national economy, especially in southern states. Methods: The study adopts a quantitative cross-sectional design with a retrospective survey carried out using the Pollfish platform in June 2022. The data were analyzed using descriptive statistics and hierarchical multiple regression to assess the three hypotheses: (H1) Chikungunya awareness is related to sociodemographic factors; (H2) Wearing long sleeves and pants is related to (a) Chikungunya awareness and (b) information-seeking behaviors, when controlling for sociodemographic variables; (H3) Use of insect repellents is related to (a) Chikungunya awareness and (b) information-seeking behaviors when controlling for sociodemographic variables. Results: The results highlight the relationships between chikungunya's awareness, information-seeking behavior, and willingness to engage in protective behaviors. 45.91% of the participants mentioned not having heard about chikungunya, and 67.07% of respondents had sought information about mosquito-borne illnesses in the past, 55.9% have looked at the U.S. State Department's website for mosquitoborne diseases, 38.32% have visited the U.S. CDC website for information specifically about chikungunya. Conclusion: The results of this study show that most American travelers are unaware of chikungunya and its mode of transmission. Travel could likely introduce the chikungunya virus to the United States. Despite increased health information-seeking behavior among U.S. residents after the Covid19 pandemic, Chikungunya awareness is low. [ABSTRACT FROM AUTHOR]

Radiocarbon (14C) in natural samples undergoes changes due to variations in atmospheric CO2 resulting from anthropogenic activities. To analyze the variation of the 14C ratio in atmospheric CO2, deciduous tree leaves were collected in Gyeongju, a popular tourist city in South Korea. Leaf samples were collected from Prunus subg. Cerasus trees at five different sampling points throughout the city over 3 years (2018, 2020, and 2021). The 14C data of the samples were categorized into three groups (downtown, rural, and tourist sites) and analyzed for variations among the different years. The 14C ratio at downtown sites was stable after 2018, the rural site ratio increased between 2018 and 2020 and then decreased between 2020 and 2021, and the tourist site ratio increased after 2018. We theorize that the increased 14C ratio at the tourist site was caused by a decrease in tourism after 2018. [ABSTRACT FROM AUTHOR]

Background: More than half of the world's population suffers from epidemic diseases that are spread by mosquitoes. The primary strategy used to stop the spread of mosquito-borne diseases is vector control. Interference RNA (RNAi) is a powerful tool for controlling insect populations and may be less susceptible to insect resistance than other strategies. However, public concerns have been raised because of the transfer of antibiotic resistance marker genes to environmental microorganisms after integration into the recipient genome, thus allowing the pathogen to acquire resistance. Therefore, in the present study, we modified the 3-hydroxykynurenine transaminase (3hkt) and hormone receptor 3 (hr3) RNAi vectors to remove antibiotic resistance marker genes and retain the expression cassette of the inverse repeat sequence of the 3hkt/hr3 target gene. This recombinant microalgal marker-free RNAi insecticide was subsequently added to the suburban water in a simulated-field trial to test its ability to control mosquito population. Methods: The expression cassette of the 3hkt/hr3 inverted repeat sequence and a DNA fragment of the argininosuccinate lyase gene without the ampicillin resistance gene were obtained using restriction enzyme digestion and recovery. After the cotransformation of Chlamydomonas, the recombinant algae was then employed to feed Aedes albopictus larvae. Ten and 300 larvae were used in small- and large-scale laboratory Ae.albopictus feeding trials, respectively. Simulated field trials were conducted using Meishe River water that was complemented with recombinant Chlamydomonas. Moreover, the impact of recombinant microalgae on phytoplankton and zooplankton in the released water was explored via high-throughput sequencing. Results: The marker-free RNAi-recombinant Chlamydomonas effectively silenced the 3hkt/hr3 target gene, resulting in the inhibition of Ae. albopictus development and also in the high rate of Ae. albopictus larvae mortality in the laboratory and simulated field trials. In addition, the results confirmed that the effect of recombinant Chlamydomonas on plankton in the released water was similar to that of the nontransgenic Chlamydomonas, which could reduce the abundance and species of plankton. Conclusions: The marker-free RNAi-recombinant Chlamydomonas are highly lethal to the Ae. albopictus mosquito, and their effect on plankton in released water is similar to that of the nontransgenic algal strains, which reduces the abundance and species of plankton. Thus, marker-free recombinant Chlamydomonas can be used for mosquito biorational control and mosquito-borne disease prevention. [ABSTRACT FROM AUTHOR]

Gene drives for mosquito population modification are novel tools for malaria control. Strategies to safely test antimalarial effectors in the field are required. Here, we modified the Anopheles gambiae zpg locus to host a CRISPR/Cas9 integral gene drive allele (zpgD) and characterized its behaviour and resistance profile. We found that zpgD dominantly sterilizes females but can induce efficient drive at other loci when it itself encounters resistance. We combined zpgD with multiple previously characterized non-autonomous payload drives and found that, as zpgD self-eliminates, it leads to conversion of mosquito cage populations at these loci. Our results demonstrate how self-eliminating drivers could allow safe testing of non-autonomous effector-traits by local population modification. They also suggest that after engendering resistance, gene drives intended for population suppression could nevertheless serve to propagate subsequently released non-autonomous payload genes, allowing modification of vector populations initially targeted for suppression. Author summary: Gene drive is a method that allows the genetic modification of entire populations of harmful organisms. Their application to tackle invasive species, agricultural pests or insect disease vectors has been suggested. For example, they could reduce the capacity of malaria mosquitoes to transmit this deadly disease to humans by producing effector molecules inhibiting the development of the Plasmodium parasite in the mosquito vector. We describe a strategy to modularize and test multiple transgenes destined for release, and to introduce only the minimal set of modifications needed into a mosquito population. We show how some elements, once no longer needed, can be made to self-eliminate from populations and we also study how several independent gene drive traits, located in different parts of the genome, can interact and propagate at the level of mosquito cage populations. [ABSTRACT FROM AUTHOR]

Direct atmospheric 14CO2 measurements began in New Zealand in 1954, initially to improve 14C as a dating tool, but quickly evolving into a method for understanding the carbon cycle. These early 14CO2 measurements immediately demonstrated the existence of an "Atom Bomb Effect," as well as an "Industrial Effect." These two gigantic tracer experiments have been utilized via 14CO2 measurements over the years to produce a wealth of knowledge in multiple research fields including atmospheric carbon cycle research, oceanography, soil science, and aging of post-bomb materials. [ABSTRACT FROM AUTHOR]

Roles of constraints in shaping evolutionary outcomes are often considered in the contexts of developmental biology and population genetics, in terms of capacities to generate new variants and how selection limits or promotes consequent phenotypic changes. Comparative genomics also recognizes the role of constraints, in terms of shaping evolution of gene and genome architectures, sequence evolutionary rates, and gene gains or losses, as well as on molecular phenotypes. Characterizing patterns of genomic change where putative functions and interactions of system components are relatively well described offers opportunities to explore whether genes with similar roles exhibit similar evolutionary trajectories. Using insect immunity as our test case system, we hypothesize that characterizing gene evolutionary histories can define distinct dynamics associated with different functional roles. We develop metrics that quantify gene evolutionary histories, employ these to characterize evolutionary features of immune gene repertoires, and explore relationships between gene family evolutionary profiles and their roles in immunity to understand how different constraints may relate to distinct dynamics. We identified three main axes of evolutionary trajectories characterized by gene duplication and synteny, maintenance/stability and sequence conservation, and loss and sequence divergence, highlighting similar and contrasting patterns across these axes amongst subsets of immune genes. Our results suggest that where and how genes participate in immune responses limit the range of possible evolutionary scenarios they exhibit. The test case study system of insect immunity highlights the potential of applying comparative genomics approaches to characterize how functional constraints on different components of biological systems govern their evolutionary trajectories. [ABSTRACT FROM AUTHOR]

When dating documents by radiocarbon (14C), what we typically measure is the concentration of the support (e.g. paper, parchment, or papyrus). This can however lead to a possible misinterpretation of results because the support may be older than the writing itself. To minimize such a possible ambiguity, the ideal approach would be direct dating of the ink or color (if organic). Here we propose a feasibility study to date carbon-based black inks when deposited on papyrus, one of the most widespread writing supports used in the past. We prepared test samples, using a commercial papyrus and a homemade black ink, obtained combining modern charcoal fragments and Arabic gum. Even though the ink binder might have represented the best candidate to be dated, we verified by FTIR that the molecular composition of its soluble fraction is very similar to papyrus extractives, thus identifying the residual charcoals recovered after extraction as the most suitable material for the measurement. Enough charcoal material was extracted from the test samples and processed using our new setup optimized for microgram-size samples. The overall experimental procedure was found to be reproducible, and measured 14C concentrations were coherent with the data obtained from larger samples and raw materials. [ABSTRACT FROM AUTHOR]

Background: Nyssorhynchus darlingi (also known as Anopheles darlingi) is the primary malaria vector in the Amazon River Basin. In Brazil, analysis of single nucleotide polymorphisms (SNPs) previously detected three major population clusters, and a common garden experiment in a laboratory setting revealed significant population variation in life history traits. Increasing temperatures and local level variation can affect life history traits, i.e. adult longevity, that alter vectorial capacity with implications for malaria transmission in Ny. darlingi. Methods: We investigated the population structure of Ny. darlingi from 7 localities across Brazil utilizing SNPs and compared them to a comprehensive Ny. darlingi catalog. To test the effects of local level variation on life history traits, we reared F1 progeny from the 7 localities at three constant temperatures (20, 24 and 28 °C), measuring key life history traits (larval development, food-starved adult lifespan, adult size and daily survival). Results: Using nextRAD genotyping-by-sequencing, 93 of the field-collected Ny. darlingi were genotyped at 33,759 loci. Results revealed three populations (K = 3), congruent with major biomes (Amazonia, Cerrado and Mata Atlântica), with greater FST values between biomes than within. In the life history experiments, increasing temperature reduced larval development time, adult lifespan, and wing length in all localities. The variation of family responses for all traits within four localities of the Amazonia biome was significant (ANOVA, P < 0.05). Individual families within localities revealed a range of responses as temperature increased, for larval development, adult lifespan, wing length and survival time. Conclusions: SNP analysis of several Brazilian localities provided results in support of a previous study wherein populations of Ny. darlingi were clustered by three major Brazilian biomes. Our laboratory results of temperature effects demonstrated that population variation in life history traits of Ny. darlingi exists at the local level, supporting previous research demonstrating the high plasticity of this species. Understanding this plasticity and inherent variation between families of Ny. darlingi at the local level should be considered when deploying intervention strategies and may improve the likelihood of successful malaria elimination in South America. [ABSTRACT FROM AUTHOR]

A radiocarbon (14C) activity analysis in the tree rings around Ignalina nuclear power plant (INPP) has been carried out with the aim to test the hypothesis to use 14C tree-ring analysis data as a tool for the reconstruction of gaseous releases from NPP to the environment. The INPP has been in decommissioning state since the end of 2009. Tree-ring samples for 14C analysis were collected 7 yr after final power unit shutdown from the INPP vicinity. The samples from 5 sampling locations were collected, prepared and measured using the Single Stage Accelerator Mass Spectrometer (SSAMS). Data analysis represents observable Ignalina NPP influence by 14C increase up to 15 pMC (percent modern carbon) in tree rings. Good correlations of the 14C concentrations and wind direction were obtained. The main purpose of this article was to match 14C measurement data along with the atmospheric dispersion modeling of emissions in order to retrospectively characterize the emission source. [ABSTRACT FROM AUTHOR]

Radiocarbon (14C) is the most accurate tracer available for quantifying atmospheric CO2 derived from fossil fuel (CO2ff), but it is expensive and time-consuming to measure. Here, we used common hourly Air Quality Index (AQI) pollutants (AQI, PM2.5, PM10, and CO) to indirectly trace diurnal CO2ff variations during certain days at the urban sites in Beijing and Xiamen, China, based on linear relationships between AQI pollutants and CO2ff traced by 14C (CO2ff−14C) for semimonthly samples obtained in 2014. We validated these indirectly traced CO2ff (CO2ff-in) concentrations against CO2ff−14C concentrations traced by simultaneous diurnal 14CO2 observations. Significant (p < 0.05) strong correlations were observed between each of the separate AQI pollutants and CO2ff−14C for the semimonthly samples. Diurnal variations in CO2ff traced by each of the AQI pollutants generally showed similar trends to those of CO2ff−14C, with high agreement at the sampling site in Beijing and relatively poor agreement at the sampling site in Xiamen. AQI pollutant tracers showed high normalized root-mean-square (NRMS) errors for the summer diurnal samples due to low CO2ff−14C concentrations. After the removal of these summer samples, the NRMS errors for AQI pollutant tracers were in the range of 31.6-64.2%. CO generally showed a high agreement and low NRMS errors among these indirect tracers. Based on these linear relationships, monthly CO2ff averages at the sampling sites in Beijing and Xiamen were traced using CO concentration as a tracer. The monthly CO2ff averages at the Beijing site showed a shallow U-type variation. These results indicate that CO can be used to trace CO2ff variations in Chinese cities with CO2ff concentrations above 5 ppm. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) is a powerful mechanism that can be exploited not only for physiology research but also for designing insect pest management approaches. Some insects cause harm by vectoring diseases dangerous to humans, livestock, or plants or by damaging crops. For at least a decade now, different insect control strategies that induce RNAi by delivering double stranded RNA (dsRNA) targeting essential genes have been proposed. Here, we focus on nontransgenic RNAi-based approaches that use oral delivery of dsRNA through feeding of inactivated bacteria to produce RNAi in disease vectors and in a crop pest. This potential pest management method could be easily adapted to target different genes or similar organisms., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Satellite imagery will offer unparalleled global spatial coverage at high-resolution for long term cost-effective monitoring of CO2 concentration plumes generated by emission hotspots. CO2 emissions can then be estimated from the magnitude of these plumes. In this paper, we assimilate pseudo-observations in a global atmospheric inversion system to assess the performance of a constellation of one to four sun-synchronous low-Earth orbit (LEO) imagers to monitor anthropogenic CO2 emissions. The constellation of imagers follows the specifications from the European Spatial Agency (ESA) for the Copernicus Anthropogenic Carbon Dioxide Monitoring (CO2M) concept for a future operational mission dedicated to the monitoring of anthropogenic CO2 emissions. This study assesses the uncertainties in the inversion estimates of emissions ("posterior uncertainties"). Results: The posterior uncertainties of emissions for individual cities and power plants are estimated for the 3 h before satellite overpasses, and extrapolated at annual scale assuming temporal auto-correlations in the uncertainties in the emission products that are used as a prior knowledge on the emissions by the Bayesian framework of the inversion. The results indicate that (i) the number of satellites has a proportional impact on the number of 3 h time windows for which emissions are constrained to better than 20%, but it has a small impact on the posterior uncertainties in annual emissions; (ii) having one satellite with wide swath would provide full images of the XCO2 plumes, and is more beneficial than having two satellites with half the width of reference swath; and (iii) an increase in the precision of XCO2 retrievals from 0.7 ppm to 0.35 ppm has a marginal impact on the emission monitoring performance. Conclusions: For all constellation configurations, only the cities and power plants with an annual emission higher than 0.5 MtC per year can have at least one 8:30–11:30 time window during one year when the emissions can be constrained to better than 20%. The potential of satellite imagers to constrain annual emissions not only depend on the design of the imagers, but also strongly depend on the temporal error structure in the prior uncertainties, which is needed to be objectively assessed in the bottom-up emission maps. [ABSTRACT FROM AUTHOR]

Background: Vector control is a key component of malaria prevention. Two major vector control strategies have been implemented in São Tomé and Príncipe (STP), indoor residual spraying (IRS) and outdoor larval control using Bacillus thuringiensis israelensis (Bti). This study evaluated post-intervention effects of control strategies on vector population density, composition, and knockdown resistance mutation, and their implications for malaria epidemiology in STP. Methods: Mosquitoes were collected by indoor and outdoor human landing catches and mosquito light traps in seven districts. Mosquito density was calculated by numbers of captured adult mosquitoes/house/working hour. Mitochondrial cytochrome c oxidase subunit I (COI) was PCR amplified and sequenced to understand the spatial–temporal population composition of malaria vector in STP. Knockdown resistance L1014F mutation was detected using allele-specific PCR. To estimate the malaria transmission risks in STP, a negative binomial regression model was constructed. The response variable was monthly incidence, and the explanatory variables were area, rainfall, entomological inoculation rate (EIR), and kdr mutation frequency. Results: Malaria vector in STP is exophilic Anopheles coluzzii with significant population differentiation between Príncipe and São Tomé (mean FST = 0.16, p < 0.001). Both vector genetic diversity and knockdown resistance mutation were relatively low in Príncipe (mean of kdr frequency = 15.82%) compared to São Tomé (mean of kdr frequency = 44.77%). Annual malaria incidence rate in STP had been rapidly controlled from 37 to 2.1% by three rounds of country-wide IRS from 2004 to 2007. Long-term application of Bti since 2007 kept the mosquito density under 10 mosquitoes/house/hr/month, and malaria incidence rate under 5% after 2008, except for a rising that occurred in 2012 (incidence rate = 6.9%). Risk factors of area (São Tomé compared to Príncipe), rainfall, outdoor EIR, and kdr mutation frequency could significantly increase malaria incidence by 9.33–11.50, 1.25, 1.07, and 1.06 fold, respectively. Conclusions: Indoor residual spraying could rapidly decrease Anopheles density and malaria incidence in STP. Outdoor larval control using Bti is a sustainable approach for controlling local vector with exophilic feature and insecticide resistance problem. Vector control interventions should be intensified especially at the north-eastern part of São Tomé to minimize impacts of outbreaks. [ABSTRACT FROM AUTHOR]

Mosquitoes play a predominant role as leading agents in the spread of vector-borne diseases and the consequent mortality in humans. Despite reports on increase of new and recurrent mosquito borne-disease outbreaks such as chikungunya, dengue fever and Rift Valley fever in Kenya, little is known about the genetic characteristics and diversity of the vector species that have been incriminated in transmission of disease pathogens. In this study, mosquito species were collected from Kisumu city, Kilifi town and Nairobi city and we determined their genetic diversity and phylogenetic relationships. PCR was used to amplify the partial cytochrome oxidase subunit 1 (CO1) gene of mosquito samples. Molecular-genetic and phylogenetic analysis of the partial cytochrome oxidase subunit 1 (CO1) gene were employed to identify their relationship with known mosquito species. Fourteen (14) haplotypes belonging to genus Aedes, nine (9) haplotypes belonging to genus Anopheles and twelve (12) haplotypes belonging to genus Culex were identified in this study. Findings from this study revealed a potentially new haplotype belonging to Anopheles genus and reported the first molecular characterization of Aedes cumminsii in Kenya. Sequence results revealed variation in mosquito species from Kilifi, Kisumu and Nairobi. Since vector competence varies greatly across species as well as species-complexes and is strongly associated with specific behavioural adaptations, proper species identification is important for vector control programs.

Mosquitoes play a predominant role as leading agents in the spread of vector-borne diseases and consequent mortality in humans. Despite reports on increase of new and recurrent mosquito borne-disease outbreaks such as chikungunya, dengue fever and Rift valley fever in Kenya little is known about the genetic characteristics and diversity of the vector species that have been incriminated in transmission of disease pathogens. In this study, we identified mosquito species across Kisumu, Kilifi and Nairobi Counties and determined their genetic diversity and phylogenetic relationships. PCR was used to amplify and sequence the partial cytochrome oxidase subunit 1 (CO1) gene of mosquito samples. Molecular-genetic and phylogenetic analysis of the partial cytochrome oxidase subunit 1 (CO1) gene was employed to identify their relationships with known mosquito species. Fourteen (14) haplotypes belonging to genus Aedes, nine (9) haplotypes belonging to genus Anopheles and twelve (12) haplotypes belonging to genus Culex were identified in this study. Findings from this study revealed a potentially new haplotype belonging to Anopheles genus and reported the first molecular characterization of Aedes cummnisii in Kenya. Sequence results revealed variation in mosquito species from Kilifi, Kisumu and Nairobi. Since vector competence varies greatly across species and species-complexes and is strongly associated with specific behavioural adaptations, proper species identification is important for vector control programs.