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Usutu virus (USUV) is a zoonotic arbovirus infecting mainly wild birds. It is transmitted by ornithophilic mosquitoes, mainly of the genus Culex from birds to birds and to several vertebrate dead-end hosts. Several USUV lineages, differing in their virulence have emerged in the last decade and now co-circulate in Europe, impacting human populations. However, their relative transmission and effects on their mosquito vectors is still not known. We thus compared the vector competence and survival of Culex pipiens mosquitoes experimentally infected with two distinct USUV lineages, EU2 and EU3, that are known to differ in their virulence and replication in vertebrate hosts. Infection rate was variable among blood feeding assays but variations between EU2 and EU3 lineages were consistent suggesting that Culex pipiens was equally susceptible to infection by both lineages. However, EU3 viral load increased with viral titer in the blood meal while EU2 viral load was high at all titers which suggest a greater replication of EU2 than EU3 in mosquito. While their relative transmission efficiencies are similar, at least at low blood meal titer, positive correlation between transmission and blood meal titer was observed for EU3 only. Contrary to published results in vertebrates, EU3 induced a higher mortality to mosquitoes (i.e. virulence) than EU2 whatever the blood meal titer. Therefore, we found evidence of lineage-specific differences in vectorial capacity and virulence to both the vector and vertebrate host which lead to balanced propagation of both viral lineages. These results highlight the need to decipher the interactions between vectors, vertebrate hosts, and the diversity of arbovirus lineages to fully understand transmission dynamics. Author summary: Usutu virus is a zoonotic arbovirus that emerged in the last decade in Europe, impacting wild bird and human populations. Several co-circulating USUV lineages differ in their virulence to vertebrate hosts but their relative transmission and effects on their mosquito vectors is still not known. We compared the vector competence and survival of Culex pipiens mosquitoes experimentally infected with two distinct USUV lineages. We showed that their relative transmission efficiencies are similar but EU3 was more virulent to mosquitoes than EU2. In addition to published results, we revealed lineage-specific differences in vectorial capacity and virulence to both the vector and the vertebrate host suggesting that propagation of both viral lineages might be balanced. Our study provides new insights into the transmission of an emerging arbovirus and highlights the need to investigate the interactions between vectors, vertebrate hosts, and the diversity of arbovirus lineages to fully understand transmission dynamics. [ABSTRACT FROM AUTHOR]

Context. West African crocodylian populations are declining and in need of conservation action. Surveys and other monitoring methods are critical components of crocodile conservation programs; however, surveys are often hindered by logistical, financial and detectability constraints. Increasingly used in wildlife monitoring programs, drones can enhance monitoring and conservation efficacy. Aims. This study aimed to determine a standard drone crocodylian survey protocol and evaluate the drones as a tool to survey the diverse crocodylian assemblage of West Africa. Methods. We surveyed crocodile populations in Benin, Côte d'Ivoire, and Niger in 2017 and 2018, by using the DJI Phantom 4 Pro drone and via traditional diurnal and nocturnal spotlight surveys. We used a series of test flights to first evaluate the impact of drones on crocodylian behaviour and determine standard flight parameters that optimise detectability. We then, consecutively, implemented the three survey methods at 23 sites to compare the efficacy of drones against traditional crocodylian survey methods. Key results. Crocodylus suchus can be closely approached (>10 m altitude) and consumer-grade drones do not elicit flight responses in West African large mammals and birds at altitudes of >40-60 m. Altitude and other flight parameters did not affect detectability, because high-resolution photos allowed accurate counting. Observer experience, field conditions (e.g. wind, sun reflection), and site characteristics (e.g. vegetation, homogeneity) all significantly affected detectability. Drone-based crocodylian surveys should be implemented from 40 m altitude in the first third of the day. Comparing survey methods, drones performed better than did traditional diurnal surveys but worse than standard nocturnal spotlight counts. The latter not only detected more individuals, but also a greater size-class diversity. However, drone surveys provide advantages over traditional methods, including precise size estimation, less disturbance, and the ability to cover greater and more remote areas. Drone survey photos allow for repeatable and quantifiable habitat assessments, detection of encroachment and other illegal activities, and leave a permanent record. Conclusions. Overall, drones offer a valuable and cost-effective alternative for surveying crocodylian populations with compelling secondary benefits, although they may not be suitable in all cases and for all species. Implications. We propose a standardised and optimised protocol for drone-based crocodylian surveys that could be used for sustainable conservation programs of crocodylians in West Africa and globally. [ABSTRACT FROM AUTHOR]

Abstract Background Analysis of Culex pipiens mosquitoes collected from a single site in Lebanon in 2005, revealed an alarming frequency of ace-1 alleles conferring resistance to organophosphate insecticides. Following this, in 2006 the majority of municipalities switched to pyrethroids after a long history of organophosphate usage in the country; however, since then no studies have assessed the impact of changing insecticide class on the frequency of resistant ace-1 alleles in C. pipiens. Methods C. pipiens mosquitoes were captured indoors from 25 villages across the country and subjected to established methods for the analysis of gene amplification at the Ester locus and target site mutations in ace-1 gene that confer resistance to organophosphates. Results We conducted the first large-scale screen for resistance to organosphosphates in C. pipiens mosquitoes collected from Lebanon. The frequency of carboxylesterase (Ester) and ace-1 alleles conferring resistance to organophosphates were assessed among C. pipiens mosquitoes collected from 25 different villages across the country between December 2008 and December 2009. Established enzymatic assay and PCR-based molecular tests, both diagnostic of the major target site mutations in ace-1 revealed the absence of the F290V mutation among sampled mosquitoes and significant reduction in the frequency of G119S mutation compared to that previously reported for mosquitoes collected from Beirut in 2005. We also identified a new duplicated ace-1 allele, named ace-1D13, exhibiting a resistant phenotype by associating a susceptible and a resistant copy of ace-1 in a mosquito line sampled from Beirut in 2005. Fisher’s exact test on ace-1 frequencies in the new sample sites, showed that some populations exhibited a significant excess of heterozygotes, suggesting that the duplicated allele is still present. Starch gel electrophoresis indicated that resistance at the Ester locus was mainly attributed to the Ester2 allele, which exhibits a broad geographical distribution. Conclusions Our analysis suggests that the frequency of resistant ace-1 alleles in mosquito populations can be downshifted, and in certain cases (F290V mutation) even eliminated, by switching to a different class of insecticides, possibly because of the fitness cost associated with these alleles.

Abstract Formal publication of the Culex genome sequence has closed the human disease vector triangle by meeting the Anopheles gambiae and Aedes aegypti genome sequences. Compared to these other mosquitoes, Culex quinquefasciatus possesses many specific hallmark characteristics, and may thus provide different angles for research which ultimately leads to a practical solution for controlling the ever increasing burden of insect-vector-borne diseases around the globe. We argue the special importance of the cosmopolitan species- Culex genome sequence by invoking many interesting questions and the possible of potential of the Culex genome to answer those.

Abstract Background A central hypothesis in the evolutionary ecology of parasitism is that trade-offs exist between resistance to parasites and other fitness components such as fecundity, growth, survival, and predator avoidance, or resistance to other parasites. These trade-offs are called costs of resistance. These costs fall into two broad categories: constitutive costs of resistance, which arise from a negative genetic covariance between immunity and other fitness-related traits, and inducible costs of resistance, which are the physiological costs incurred by hosts when mounting an immune response. We sought to study inducible costs in depth using the crustacean Daphnia magna and its bacterial parasite Pasteuria ramosa. Results We designed specific experiments to study the costs induced by exposure to this parasite, and we re-analysed previously published data in an effort to determine the generality of such costs. However, despite the variety of genetic backgrounds of both hosts and parasites, and the different exposure protocols and environmental conditions used in these experiment, this work showed that costs of exposure can only rarely be detected in the D. magna-P. ramosa system. Conclusions We discuss possible reasons for this lack of detectable costs, including scenarios where costs of resistance to parasites might not play a major role in the co-evolution of hosts and parasites.

Abstract Background Insecticide resistance is a rapid and recent evolutionary phenomenon with serious economic and public health implications. In the mosquito Anopheles gambiae s.s., main vector of malaria, resistance to organophosphates and carbamates is mainly due to a single amino-acid substitution in acetylcholinesterase 1 (AChE1). This mutation entails a large fitness cost. However, a resistant duplicated allele of the gene encoding AChE1 (ace-1), potentially associated to a lower fitness cost, recently appeared in An. gambiae. Methods Using molecular phenotype data collected from natural populations from West Africa, the frequency of this duplicated allele was investigated by statistical inference. This method is based on the departure from Hardy-Weinberg phenotypic frequency equilibrium caused by the presence of this new allele. Results The duplicated allele, Ag-ace-1D, reaches a frequency up to 0.65 in Ivory Coast and Burkina Faso, and is potentially present in Benin. A previous study showed that Ag-ace-1D, present in both M and S molecular forms in different West Africa countries, was generated by a single genetic event. This single origin and its present distribution suggest that this new allele is currently spreading. Conclusion The spread of this less costly resistance allele could represent a major threat to public health, as it may impede An. gambiae control strategies, and thus increases the risk of malaria outbreaks.

BACKGROUND: Mosquitoes of the Culex pipiens complex are the vectors of several arboviruses and are thus subjected to insecticide control worldwide. However, overuse of insecticides selects for resistance. While assessing the resistance status of the vectors is required for effective and sustainable disease control, resistance has so far only been sparsely studied in Morocco. In this study, we establish a first countrywide assessment of the levels of resistance to various insecticides and the potential responsible mechanisms involved. Cx. pipiens larvae were collected from natural populations of five regions of Morocco, and their taxonomic status was determined (molecular forms). The level of their susceptibility to insecticides was assessed by single‐diagnostic‐dose bioassays. Molecular identification of known resistance alleles was investigated to determine the frequency of target‐site mutations. RESULTS: This study confirms that Moroccan populations are an interbreeding mix of pipiens and molestus forms, with large gene flow for the resistance alleles. We also found that Cx. pipiens mosquitoes are resistant to all insecticide families, all over Morocco: resistance is high for insecticides used in mosquito control, but also present for other pesticides. Resistance alleles are similarly more frequent for mosquito control insecticides. However, their distribution is heterogeneous in the five regions, with significant genetic differentiation between populations, revealing the crucial role of local insecticide treatment practices. CONCLUSION: This study provides reference countrywide data that highlight the need for further research to refine the distribution of resistance in Morocco and to understand the role of agriculture/urban residuals in its spread. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND: Arboviruses are controlled through insecticide control of their mosquito vector. However, inconsiderate use of insecticides often results in the selection of resistance in treated populations, so that monitoring is required to optimize their usage. Here, Culex pipiens (West Nile and Rift Valley Fever virus vector) specimens were collected from four Moroccan cities. Levels of susceptibility to the organophosphate (OP) insecticide malathion were assessed using World Health Organization (WHO)‐recommended bioassays. Individual mosquitoes were tested for the presence of the G119S mutation in the ace‐1 gene, the main OP‐target resistance mutation. RESULTS: Bioassays showed that mosquitoes from Mohammedia were significantly more resistant to malathion than those from Marrakech. Analyzing the ace‐1 genotypes in dead and surviving individuals suggested that other resistance mechanisms may be present in Mohammedia. The ace‐1 resistance allele frequencies were relatively moderate (< 0.4). Their analyses in three Moroccan cities (Tangier, Casablanca and Marrakech) however showed disparities between two coexisting Cx. pipiens forms and revealed that the G119S mutation tends to be more frequent in urban than in rural collection sites. CONCLUSION: These findings provide a reference assessment of OP resistance in Morocco and should help the health authorities to develop informed and sustainable vector control programs. © 2018 Society of Chemical Industry Recurrent vector control selects for insecticide resistance. This article presents an evaluation of target‐mutation resistance to organophosphate insecticides in the Culex pipiens complex in Morocco, and its relation to urbanization. [ABSTRACT FROM AUTHOR]