Biology subjects

Dusfour, I.

Publications and source records attributed to Dusfour, I..

13 recordsLinked to original sources

bioRxiv2025

Bridging laboratory and field research: method adjustments to manipulate field-derived Aedes aegypti

Reference strains of Aedes aegypti, reared over numerous generations under laboratory conditions, are selected to fit laboratory conditions and commonly used in research due to their consistency and ease of handling. However, working exclusively with such strains may not fully reflect the traits of natural populations. While working on such strains is relevant in terms of reproducibility between labs, it is also important to perform some work on field-collected mosquitoes and their progeny to capture more representative biological and behavioural variation. In this study, we used the New Orleans reference strain as a control to adjust and evaluate methods to manipulate the F1 progeny of field-collected mosquitoes from Cayenne (French Guiana). To improve blood feeding rate, we tested the impact of several blood feeding systems for mosquitoes kept in a cage or in individual vials and adjusted starvation time. To monitor fertilization, we assessed if dissection buffer affects burst of spermatheca during dissection, whether mosquitoes were collected alive of shortly after death. The results described here may be helpful for studies on mosquito fitness, particularly on field-derived mosquitoes or on experiments requiring individual level monitoring. Author summaryAedes aegypti is the major vector of dengue virus, causing hundreds of millions of cases per year. To fight against disease transmission, mosquitoes are studied in the laboratory, notably using colonies of mosquitoes kept over decades in the laboratory. These reference strains have been selected to fit laboratory conditions and may thus not fully reflect the traits of natural populations. While working on such strains is relevant in a sake of consistency between labs and ease of handling, it is also important to work on field-collected mosquitoes and their progeny to capture more representative biological and behavioural variation. In this study, we adjusted and evaluated methods to manipulate the F1 progeny of field-collected mosquitoes from Cayenne (French Guiana), using the New Orleans reference strain as a control. To improve blood feeding rate, we tested the impact of several blood feeding systems for mosquitoes kept in a cage or in individual vials and adjusted starvation time. To monitor fertilization, we assessed if dissection buffer affects burst of spermatheca during dissection, whether mosquitoes were collected alive of shortly after death. The results described here may be helpful for studies on mosquito fitness, particularly on field-derived mosquitoes or on experiments requiring individual level monitoring.

zoology↗

bioRxiv2024

A DNA barcode library for Culex mosquitoes (Diptera: Culicidae) of South America with the description of two cryptic species of subgenus Melanoconion

The genus Culex is one of the most diverse in the world and includes numerous known vector species of parasites and viruses to humans. Morphological identification of Culex species is notoriously difficult and rely mostly on the examination of properly dissected male genitalia which largely prevents female and immature identification during entomological, ecological or arboviral surveys. The aims of this study were (i) to establish a DNA barcode library for Culex mosquitoes of French Guiana based on the mitochondrial gene cytochrome c oxidase I (COI) marker, (ii) to compare three approaches of molecular delimitation of species to morphological identification, and (iii) to test the effectiveness of the COI marker at a broader geographical scale across South America. Mosquitoes used in this study were sampled in French Guiana between 2013 and 2023. We provide 246 COI sequences for 90 morphologically identified species of Culex, including five new country records and two newly described species. Overall, congruence between morphological identification and molecular delimitations using the COI barcode were high. The Barcode of Life Data clustering approach into Barcode Index Numbers gives the best result in terms of species delimitation, followed by the muti-rate Poisson Tree Processes and the Assemble Species by Automatic Partitioning methods. Inconsistencies between morphological identification and molecular delimitation can be explained by introgression, incomplete lineage sorting, imperfect taxonomy or the effect of the geographical scale of sampling. This increases by almost two-fold the number of mosquito species for which a DNA barcode is available in French Guiana, including 75% of the species of Culex currently known in the territory. Finally, this study confirms the usefulness of the COI barcode in identifying Culex mosquitoes of South America, but also points the limits of this marker for some groups of species within the subgenera Culex and Melanoconion.

molecular biology↗

bioRxiv2024

A genomic duplication spanning multiple P450s contributes to insecticide resistance in the dengue mosquito Aedes aegypti

Resistance of mosquitoes to insecticides is one example of rapid adaptation to anthropogenic selection pressures having a strong impact on human health and activities. Target-site modification and increased insecticide detoxification are the two main mechanisms underlying insecticide resistance in mosquitoes. While target-sites mutations are well characterised and often used to track resistance in the field, the genomic events associated with insecticide detoxification remain partially characterised. Recent studies evidenced the key role of gene duplications in the over-expression of detoxification enzymes and their potential use to track metabolic resistance alleles in the field. However, such genomic events remain difficult to characterise due to their complex genomic architecture and their co-occurrence with other resistance alleles. In this concern, the present work investigated the role of a large genomic duplication affecting a cluster of detoxification enzymes in conferring resistance to the pyrethroid insecticide deltamethrin in the mosquito Aedes aegypti. Two isofemale lines originating from French Guiana and being deprived from major target-site mutations showed distinct insecticide resistance levels. Combining RNA-seq and whole genome pool-seq identified a 220 Kb genomic duplication enhancing the expression of multiple contiguous cytochrome P450s in the resistant line. The genomic architecture of the duplicated loci was elucidated through long read sequencing, evidencing its transposon-mediated evolutionary origin. The involvement of this P450 duplication in deltamethrin survival was supported by a significant phenotypic response to the P450 inhibitor piperonyl butoxide together with genotype-phenotype association and RNA interference. Experimental evolution suggested that this P450 duplication is associated with a significant fitness cost, potentially affecting its adaptive value in presence of other resistance alleles. Overall, this study supports the importance of genomic duplications affecting detoxification enzymes in the rapid adaptation of mosquitoes to insecticides. Deciphering their genomic architecture provides new insights into the evolutionary processes underlying such rapid adaptation. Such findings provide new tools for the surveillance and management of resistance in the field.

evolutionary biology↗

bioRxiv2023

Potential of MALDI-TOF MS biotyping to detect deltamethrin resistance in the dengue vector Aedes aegypti.

Insecticide resistance in mosquitoes is spreading worldwide and represents a growing threat to vector control. Insecticide resistance is caused by different mechanisms including higher metabolic detoxication, target-site modification, reduced penetration and behavioral changes that are not easily detectable with simple diagnostic methods. Indeed, most molecular resistance diagnostic tools are costly and labor intensive and then difficult to use for routine monitoring of insecticide resistance. The present study aims to determine whether mosquito susceptibility status against the pyrethroid insecticides (mostly used for mosquito control) could be established by the protein signatures of legs and/or thoraxes submitted to MALDI-TOF Mass Spectrometry (MS). The quality of MS spectra for both body parts was controlled to avoid any bias due to unconformity protein profiling. The comparison of MS profiles from three inbreeds Ae. aegypti lines from French Guiana (IRF, IR03, IR13), with distinct deltamethrin resistance genotype / phenotype and the susceptible reference laboratory line BORA (French Polynesia), showed different protein signatures. On both body parts, the analysis of whole protein profiles revealed a singularity of BORA line compared to the three inbreeding lines from French Guiana origin, suggesting that the first criteria of differentiation is the geographical origin and/or the breeding history rather than the insecticide susceptibility profile. However, a deeper analysis of the protein profiles allowed to identify 10 and 11 discriminating peaks from leg and thorax spectra, respectively. Among them, a specific peak around 4870 Da was detected in legs and thoraxes of pyrethroid resistant lines compared to the susceptible counterparts hence suggesting that MS profiling may be promising to rapidly distinguish resistant and susceptible phenotypes. Further work is needed to confirm the nature of this peak as a deltamethrin resistant marker and to validate the routine use of MS profiling to track insecticide resistance in Ae. aegypti field populations. Author SummaryThe monitoring of mosquito insecticide resistance in local populations is essential to guide the choice of the vector control strategy. Current methods for resistance monitoring rely on biological, biochemical and molecular assays that all have their weakness. To circumvent these limitations, alternative methods have to be explored. In previous studies, MALDI-TOF MS profiling have proved it performance to classify mosquitoes at the species and sub-species levels. The present work aim was to assess whether MALDI-TOF MS profiling strategy could be useful for determination of mosquito susceptibility to the most used pyrethroid insecticide. In this way, four mosquito lines with distinct deltamethrin resistance genotype / phenotype were submitted to MS analysis. The accurate comparison of MS spectra showed different peak intensities between mosquitoes exhibiting different insecticide resistance profiles. Among discriminant peaks, one may be promising to detect insecticide-resistance mechanisms in public health mosquitoes. A better characterization of mosquito life traits will help countries to implement timely and locally adapted vector control interventions.

bioengineering↗

bioRxiv2022

Relevance study of vector competence and insecticide resistance in Aedes aegypti laboratory lines

The urban mosquito species Aedes aegypti is the main vector of arboviruses worldwide. Mosquito control with insecticides is the most prevalent method for preventing transmission in the absence of effective vaccines and available treatments; however, the extensive use of insecticides has led to the development of resistance in mosquito populations throughout the world, and the number of epidemics caused by arboviruses has increased. Three mosquito lines with different resistance profiles to deltamethrin were isolated in French Guiana, including one with the I1016 knock-down resistant allele. Significant differences were observed in the cumulative proportion of mosquitoes with a disseminated chikungunya virus infection over time. In addition, certain genes (CYP6BB2, CYP6N12, GST2, trypsin) were variably overexpressed in the midgut at 7 days after an infectious blood meal in these three lines. Therefore, detoxification enzymes and kdr mutations may contribute to an enhanced midgut barrier and reduced dissemination rate. Our work shows that vector competence for chikungunya virus varied between Ae. aegypti laboratory lines with different deltamethrin-resistance profiles. More accurate verification of the functional association between insecticide resistance and vector competence remains to be demonstrated. ImportanceThree Ae. aegypti lines, isolated from the same collection site, underwent different insecticide selection pressures against deltamethrin under laboratory conditions. As a result, they developed different resistant profiles. In this study, when these lines were fed an artificial infectious blood meal containing chikungunya virus, all three lines including the reference strain showed a high infection rate. There was no statistical difference in infection rate found; however, the dissemination rate of the virus from midgut to head were significantly different. A higher resistance level detected by the WHO test was correlated with a lower viral dissemination rate for each strain. This study presented evidence that the insecticide selection pressure or the existence of insecticide resistance could lead to differences in viral dissemination or even transmission in mosquito populations. We hope that our study can give more insights into understanding the roles of mosquito insecticide resistance on viral transmission.

molecular biology↗

bioRxiv2021

Quantification of breeding sites parameters in shaping bacterial communities in Aedes aegypti (Diptera: Culicidae).

Studies have demonstrated the importance of breeding site, few had disentangled the role of microbiome, physico-chemical and biological factors of water as well as landuse on larval microbial communities and their recruitment in mosquito. A quantitative exploration of the interplay of multiple factors on mosquito microbiome was performed using a dataset obtained through a field survey undertaken in French Guiana. Two complementary hypotheses were tested (i) the most dissimilar larval microbiome structures in breeding sites displayed the most contrasting water properties and land-use, (ii) a higher specificity level of environmental parameters have an incidence on larval microbiome. Variance partitioning approach validated the two hypothesis by providing evidence that water bacterial community is a most significant driver shaping the structure of the bacteriome in mosquito than other environmental parameters from the breeding sites. However, land-use does not play such important role to explain variance. Our results consolidate and complement the knowledge shaping mosquito microbiota but also highlighted the large unknown in understanding the ecology of the recruitment into host.

ecology↗

bioRxiv2021

New Records of California Serogroup Virus in Aedes Mosquitoes and First Detection in Simulioidae Flies from Northern Canada and Alaska

An expected consequence of climate warming is an expansion of the geographic distribution of biting insects and associated arthropod-borne diseases (arboviruses). Emerging and reemerging arboviruses that can affect human health are likely to pose significant consequences for Northern communities where access to health resources is limited. In the North American Arctic, little is known about arboviruses. Thus, in 2019, we sampled biting insects in Nunavik (Kuujjuaq), Nunavut (Igloolik, Karrak Lake and Cambridge Bay), Northwest Territories (Igloolik and Yellowknife) and Alaska (Fairbanks). The main objective was to detect the presence of California serogroup viruses (CSGv) - a widespread group of arboviruses across North America and that is known to cause a wide range of symptoms, ranging from mild febrile illness to fatal encephalitis. Biting insects were captured twice daily for a 7-day period in mid-summer, using a standardized protocol consisting of 100 figure-eight movements of a sweep net. Captured specimens were separated by genus (mosquitoes) or by superfamily (other insects), and then grouped into pools of 75 by geographical locations. In total, 5079 Aedes mosquitoes and 1014 Simulioidae flies were caught. We report the detection of CSGv RNA in mosquitoes captured in Nunavut (Karrak Lake) and Nunavik (Kuujjuaq). We also report, for the first time in North America, the presence of CSGv RNA in Simulioidae flies. These results highlight the potential of biting insects for tracking any future emergence of arboviruses in the North, thereby providing key information for public health in Northern communities.

microbiology↗

bioRxiv2020

Variation in pyrethroid resistance phenotypes in Anopheles darlingi from residual malaria transmission area: warning on suspected resistance built-up in French Guiana.

Anopheles darlingi is the major vector of malaria in South America. In French Guiana, malaria transmission occurs inland and along the rivers with a particular reemergence in the lower Oyapock area. Control against malaria vector is performed using deltamethrin indoor residual spraying and long lasting impregnated bednets. For four years, the level of resistance to pyrethroids was monitored using CDC bottle tests in An. darlingi populations. Resistance built-up was suspected in a mosquito population in malaria endemic area but did not sustained, supposably due to the reintroduction of susceptible alleles. No mutation on the insecticide target genes was found, metabolic resistance is then suspected.

zoology↗

bioRxiv2020

Identification of French Guiana anopheline mosquitoes by MALDI-TOF MS profiling using protein signatures from two body parts.

In French Guiana, the malaria, a parasitic infection transmitted by Anopheline mosquitoes, remains a disease of public health importance. To prevent malaria transmission, the main effective way remains Anopheles control. For an effective control, accurate Anopheles species identification is indispensable to distinguish malaria vectors from non-vectors. Although, morphological and molecular methods are largely used, an innovative tool, based on protein pattern comparisons, the Matrix Assisted Laser Desorption / Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) profiling, emerged this last decade for arthropod identification. However, the limited mosquito fauna diversity of reference MS spectra remains one of the main drawback for it large usage. The aim of the present study was then to create and to share reference MS spectra for the identification of French Guiana Anopheline species. A total of eight distinct Anopheles species, among which four are malaria vectors, were collected in 6 areas. To improve Anopheles identification, two body parts, legs and thoraxes, were independently submitted to MS for the creation of respective reference MS spectra database (DB). This study underlined that double checking by MS enhanced the Anopheles identification confidence and rate of reliable classification. The sharing of this reference MS spectra DB should made easier Anopheles species monitoring in endemic malaria area to help malaria vector control or elimination programs.

microbiology↗

bioRxiv2019

A new high-throughput tool to screen mosquito-borne viruses in Zika virus endemic/epidemic areas

Mosquitoes are vectors of arboviruses affecting animal and human health. Arboviruses circulate primarily within an enzootic cycle and recurrent spillovers contribute to the emergence of human-adapted viruses able to initiate an urban cycle involving anthropophilic mosquitoes. The increasing volume of travel and trade offers multiple opportunities for arbovirus introduction in new regions. This scenario has been exemplified recently with the Zika pandemic. To incriminate a mosquito as vector of a pathogen, several criteria are required such as the detection of natural infections in mosquitoes. In this study, we used a high-throughput chip based on the BioMarkTM Dynamic arrays system capable of detecting 64 arboviruses in a single experiment. A total of 17,958 mosquitoes collected in Zika-endemic/epidemic countries (Brazil, French Guiana, Guadeloupe, Suriname, Senegal, and Cambodia) were analyzed. Here we show that this new tool can detect endemic and epidemic viruses in different mosquito species in an epidemic context. Thus, this fast and low-cost method can be suggested as a novel epidemiological surveillance tool to identify circulating arboviruses.

molecular biology↗

bioRxiv2019

Combining genetic crosses and pool targeted DNA-seq for untangling genomic variations associated with resistance to multiple insecticides in the dengue vector Aedes aegypti

In addition to combating vector-borne diseases, studying the adaptation of mosquitoes to insecticides provides a remarkable example of evolution-in-action driving the selection of complex phenotypes. Indeed, most resistant mosquito populations show multi-resistance phenotypes as a consequence of the variety of insecticides employed and of the complexity of selected resistance mechanisms. Such complexity makes challenging the identification of alleles conferring resistance to specific insecticides and prevents the development of molecular assays to track them in the field. Here we showed that combining simple genetic crosses with pool targeted DNA-seq can enhance the specificity of resistance alleles detection while maintaining experimental work and sequencing effort at reasonable levels. A multi-resistant population of the mosquito Aedes aegypti was exposed to three distinct insecticides (deltamethrin, bendiocarb and fenitrothion) and survivors to each insecticide were crossed with a susceptible strain to generate 3 distinct lines. F2 individuals from each line were then segregated with 2 insecticide doses. Bioassays supported the improved segregation of resistance alleles between lines. Hundreds of genes covering all detoxifying enzymes and insecticide targets together with more than 7,000 intergenic regions equally spread over mosquito genome were sequenced from pools of F0 and F2 individuals unexposed or surviving insecticide. Differential coverage analysis identified 39 detoxification enzymes showing an increased gene copy number in association with resistance. Combining an allele frequency filtering approach with a Bayesian FST-based genome scan identified multiple genomic regions showing strong selection signatures together with 50 non-synonymous variations associated with resistance. This study provides a simple and cost-effective approach to improve the segregation of resistant alleles in multi-resistant populations while reducing false positives frequently arising when comparing populations showing divergent genetic backgrounds. The identification of these insecticide resistance markers paves the way for the design of novel DNA-based resistance tracking assays.\n\nAuthor summaryIn addition to combating vector-borne diseases, understanding how mosquitoes adapt to insecticides provides a remarkable example of evolution-in-action. However, the variety of insecticides used and the complexity of adaptive mechanisms make it difficult to identify the genetic changes conferring resistance to each insecticide. Here we combined simple controlled crosses with mass DNA sequencing for enhancing the specificity of resistance gene detection. A multi-resistant mosquito population was exposed to three distinct insecticides and survivors were crossed with a susceptible strain to generate 3 distinct mosquito lines. Individuals from the second generation of each line were then segregated based on their resistance to each insecticide. Bioassays supported the improved segregation of genetic resistance markers between lines. Hundreds of genes potentially involved in resistance together with thousands non-genic regions equally spread over mosquito genome were sequenced from individuals from each line. Genomic analyses identified detoxification enzymes showing an increased gene copy number in association with resistance and multiple genomic regions showing strong selection signatures and carrying point mutations associated with resistance. Such approach improves the specificity of resistance gene detection in field mosquito populations resisting to multiple insecticides and paves the way for the design of novel DNA-based resistance tracking tools.

genomics↗

bioRxiv2018

Worldwide survey reveals lower susceptibility of African Aedes aegypti mosquitoes to diverse strains of Zika virus

Zika virus (ZIKV) is a flavivirus mainly transmitted to humans through the bite of infected Aedes aegypti mosquitoes. First isolated in Uganda in 1947, ZIKV was shown to circulate in enzootic sylvatic cycles in Africa and Asia for at least half a century before the first reported human epidemic occurred in 2007 on the Pacific island of Yap, Micronesia. Subsequently, larger ZIKV outbreaks were recorded in French Polynesia and other South Pacific islands during 2013-2014. In 2015, ZIKV reached Brazil from where it rapidly spread across the Americas and the Caribbean, causing hundreds of thousands of human cases. The factors that have fueled the explosiveness and magnitude of ZIKV emergence in the Pacific and the Americas are poorly understood. Reciprocally, the lack of major human epidemics of ZIKV in regions with seemingly favorable conditions, such as Africa or Asia, remains largely unexplained. To evaluate the potential contribution of vector population diversity to ZIKV epidemiological patterns, we established dose-response curves for eight field-derived Ae. aegypti populations representing the global range of the species, following experimental exposure to six low-passage ZIKV strains spanning the current viral genetic diversity. Our results reveal that African Ae. aegypti are significantly less susceptible than non-African Ae. aegypti across all ZIKV strains tested. We suggest that low susceptibility of vector populations may have contributed to prevent large-scale human transmission of ZIKV in Africa.

epidemiology↗

bioRxiv2017

Diverse laboratory colonies of Aedes aegypti harbor the same adult midgut bacterial microbiome

BackgroundHost-associated microbes, collectively known as the microbiota, play an important role in the biology of multicellular organisms. In mosquito vectors of human pathogens, the gut bacterial microbiota influences vectorial capacity and has become the subject of intense study. In laboratory studies of vector biology, genetic effects are often inferred from differences between geographically and genetically diverse colonies of mosquitoes that are reared in the same insectary. It is unclear, however, to what extent genetic effects can be confounded by uncontrolled differences in the microbiota composition among mosquito colonies. To address this question, we used 16S metagenomics to compare the midgut bacterial microbiome of six recent laboratory colonies of Aedes aegypti representing the geographical range and genetic diversity of the species.\n\nResultsWe found that the diversity, abundance, and community structure of the midgut bacterial microbiome was remarkably similar among the six different colonies of Ae. aegypti, regardless of their geographic origin. We also confirmed the relatively low complexity of bacterial communities inhabiting the mosquito midgut.\n\nConclusionsOur finding that geographically diverse colonies of Ae. aegypti reared in the same insectary harbor a similar gut bacterial microbiome supports the conclusion that the gut microbiota of adult mosquitoes is environmentally determined regardless of the host genotype. Thus, uncontrolled differences in microbiota composition are unlikely to represent a significant confounding factor in genetic studies of vector biology.

genetics↗