Accurate identification of Culicidae at aquatic developmental stages by MALDI-TOF MS profiling

International audienceBACKGROUND: The identification of mosquito vectors is generally based on morphological criteria, but for aquatic stages, morphological characteristics may be missing, leading to incomplete or incorrect identification. The high cost of molecular biology techniques requires the development of an alternative strategy. In the last decade, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling has proved to be efficient for arthropod identification at the species level.METHODS:To investigate the usefulness of MALDI-TOF MS for the identification of mosquitoes at aquatic stages, optimizations of sample preparation, diet, body parts and storage conditions were tested. Protein extracts of whole specimens from second larval stage to pupae were selected for the creation of a reference spectra database. The database included a total of 95 laboratory-reared specimens of 6 mosquito species, including Anopheles gambiae (S form), Anopheles coluzzi (M form), Culex pipiens pipiens, Culex pipiens molestus, Aedes aegypti and 2 colonies of Aedes albopictus.RESULTS:The present study revealed that whole specimens at aquatic stages produced reproducible and singular spectra according to the mosquito species. Moreover, MS protein profiles appeared weakly affected by the diet provided. Despite the low diversity of some MS profiles, notably for cryptic species, clustering analyses correctly classified all specimens tested at the species level followed by the clustering of early vs. late aquatic developmental stages. Discriminant mass peaks were recorded for the 6 mosquito species analyzed at larval stage 3 and the pupal stage. Querying against the reference spectra database of 149 new specimens at different aquatic stages from the 6 mosquito species revealed that 147 specimens were correctly identified at the species level and that early and late developmental stages were also distinguished.CONCLUSIONS:The present work highlights that MALDI-TOF MS profiling may be useful for the rapid and reliable identification of mosquito species at aquatic stages. With this proteomic tool, it becomes now conceivable to survey mosquito breeding sites prior to the mosquitoes' emergence and to adapt anti-vectorial measures according to the mosquito fauna detected

Raman Identification of Pigments in Wall Paintings of the Colonial Period from Bolivian Churches in the Ruta De La Plata

Micro-Raman spectroscopy was used to analyze and identify pigments and determine the palette of color in wall paintings from four churches of the Colonial Period, in the Bolivian highlands. The ancient churches Santiago de Callapa, Curahuara de Carangas, NuestraSeñora de Copacabana de Andamarca and San José de Soracachi were constructed during the 17th and 18th centuries, around the important colonial Ruta de la Plata, between Potosí city and Arica harbor on the Pacific coast of South America. Most pigments used in the wall paintings correspond to inorganic minerals as well as to organic colorants. Fully chlorinated copper phthalocyanine (green), copper phthalocyanine (blue), β-naphthol (yellow) and benzoimidazolone (red), were identified in the corresponding colored areas suggesting subsequent recent interventions or restorations. Ground layer materials were also characterized. The present results allow for discussion of the pictorial techniques used in the Andean highland during the colonial period and allow conservators and restorers to make important decisions regarding the type of intervention to perform in the case of these important historical and patrimonial monuments

Estrategias pedagógicas para el aprendizaje de la gestión integral de los residuos sólidos peligrosos del Barrio José Prudencio Padilla en el Municipio de Urumita, La Guajira

The purpose of the study was to reveal the pedagogical strategies for learning the integral management of hazardous solid waste in the José Prudencio Padilla neighborhood of the municipality of Urumita, La Guajira, Colombia. Its qualitative descriptive methodology, its scope was to work with the protagonists of the research that will have an important role, since they are the ones who will make known the practices and realities that are generated within this context and that will bring us closer to the response of the investigative questions. The population of this study will be 5 inhabitants, contemplated by the community action board made up of a president, a vice president, a secretary, a treasurer and a prosecutor, as a technique and data collection instruments, the survey is used as an observation technique and the questionnaire of 19 closed questions as an instrument. It is concluded that it is important to consider the integration of hazardous waste management as a task to avoid affecting different sectors of society, since many measures must be taken to ensure that this waste is adequately treated, used and disposed of, therefore, methods, programs, activities, structures and organizations. it must be coordinated, involving the public in the process of an important role in decision-making and the search for alternative solutions.El propósito del estudio fue Develar las estrategias pedagógicas para el aprendizaje de la gestión integral de residuos sólidos peligrosos en el barrio José Prudencio Padilla del municipio de Urumita, la Guajira, Colombia. Su metodología cualitativa tipo descriptiva, su alcance fue trabajar con los protagonistas de la investigación que tendrán un papel importante, puesto que son ellos quienes darán a conocer las prácticas y las realidades que se gestan dentro de este contexto y que nos acercaran a la respuesta de las preguntas investigativa. La población de este estudio será de 5 habitantes, contemplada por la junta de acción comunal integrada por un presidente, un vicepresidente, un secretario, un tesorero y un fiscal, como técnica e instrumentos de recolección de datos emplea la encuesta como técnica de observación y el cuestionario de 19 preguntas cerradas como instrumento. Se concluye que es importante considerar la integración de gestión de los residuos peligrosos como tarea para evitar la afectación a diferentes sectores de la sociedad, ya que se deben tomar muchas medidas para asegurar que estos residuos sean adecuadamente tratados, aprovechados y dispuestos, por tanto, los métodos programas, actividades, estructuras y organizaciones. debe ser coordinado, involucrando al público en el proceso de un papel importante en la toma de decisiones y la búsqueda de soluciones alternativas

Basic Reproduction Number of Chikungunya Virus Transmitted by Aedes Mosquitoes

We estimated the weighted mean basic reproduction number (R0) of chikungunya virus based on outbreak size. R0 was 3.4 (95% CI 2.4–4.2) and varied for 2 primary chi- kungunya mosquito vectors: 4.1 (95% CI 1.5–6.6) for Ae- des aegypti and 2.8 (95% CI 1.8–3.8) for Ae. albopictu

Biochemical and structural characterization of a novel thermophilic esterase EstD11 provide catalytic insights for the HSL family

[Abstract]: A novel esterase, EstD11, has been discovered in a hot spring metagenomic library. It is a thermophilic and thermostable esterase with an optimum temperature of 60 C. A detailed substrate preference analysis of EstD11 was done using a library of chromogenic ester substrate that revealed the broad substrate specificity of EstD11 with significant measurable activity against 16 substrates with varied chain length, steric hindrance, aromaticity and flexibility of the linker between the carboxyl and the alcohol moiety of the ester. The tridimensional structures of EstD11 and the inactive mutant have been determined at atomic resolutions. Structural and bioinformatic analysis, confirm that EstD11 belongs to the family IV, the hormone-sensitive lipase (HSL) family, from the α/β-hydrolase superfamily. The canonical a/b hydrolase domain is completed by a cap domain, composed by two subdomains that can unmask of the active site to allow the substrate to enter. Eight crystallographic complexes were solved with different substrates and reaction products that allowed identification of the hot-spots in the active site underlying the specificity of the protein. Crystallization and/or incubation of EstD11 at high temperature provided unique information on cap dynamics and a first glimpse of enzymatic activity in vivo. Very interestingly, we have discovered a unique Met zipper lining the active site and the cap domains that could be essential in pivotal aspects as thermo-stability and substrate promiscuity in EstD11Ministerio de Ciencia e Innovación; BFU2017-90030-

Chikungunya Virus Transmission Potential by Local Aedes Mosquitoes in the Americas and Europe

Citation: Vega-Rua, A., Lourenco-de-Oliveira, R., Mousson, L., Vazeille, M., Fuchs, S., Yebakima, A., . . . Failloux, A. B. (2015). Chikungunya Virus Transmission Potential by Local Aedes Mosquitoes in the Americas and Europe. Plos Neglected Tropical Diseases, 9(5), 18. doi:10.1371/journal.pntd.0003780Background Chikungunya virus (CHIKV), mainly transmitted in urban areas by the mosquitoes Aedes aegypti and Aedes albopictus, constitutes a major public health problem. In late 2013, CHIKV emerged on Saint-Martin Island in the Caribbean and spread throughout the region reaching more than 40 countries. Thus far, Ae. aegypti mosquitoes have been implicated as the sole vector in the outbreaks, leading to the hypothesis that CHIKV spread could be limited only to regions where this mosquito species is dominant. Methodology/Principal Findings We determined the ability of local populations of Ae. aegypti and Ae. albopictus from the Americas and Europe to transmit the CHIKV strain of the Asian genotype isolated from Saint-Martin Island (CHIKV_SM) during the recent epidemic, and an East-Central-South African (ECSA) genotype CHIKV strain isolated from La Reunion Island (CHIKV_LR) as a well-characterized control virus. We also evaluated the effect of temperature on transmission of CHIKV_SM by European Ae. albopictus. We found that (i) Aedes aegypti from Saint-Martin Island transmit CHIKV_SM and CHIKV_LR with similar efficiency, (ii) Ae. aegypti from the Americas display similar transmission efficiency for CHIKV_SM, (iii) American and European populations of the alternative vector species Ae. albopictus were as competent as Ae. aegypti populations with respect to transmission of CHIKV_SM and (iv) exposure of European Ae. albopictus to low temperatures (20 degrees C) significantly reduced the transmission potential for CHIKV_SM. Conclusions/Significance CHIKV strains belonging to the ECSA genotype could also have initiated local transmission in the new world. Additionally, the ongoing CHIKV outbreak in the Americas could potentially spread throughout Ae. aegypti-and Ae. albopictus-infested regions of the Americas with possible imported cases of CHIKV to Ae. albopictus-infested regions in Europe. Colder temperatures may decrease the local transmission of CHIKV_SM by European Ae. albopictus, potentially explaining the lack of autochthonous transmission of CHIKV_SM in Europe despite the hundreds of imported CHIKV cases returning from the Caribbean.Additional Authors: de Lamballerie, X.;Failloux, A. B

Shifting From Sectoral to Integrated Surveillance by Changing Collaborative Practices: Application to West Nile Virus Surveillance in a Small Island State of the Caribbean

After spreading in the Americas, West Nile virus was detected in Guadeloupe (French West Indies) for the first time in 2002. Ever since, several organizations have conducted research, serological surveys, and surveillance activities to detect the virus in horses, birds, mosquitoes, and humans. Organizations often carried them out independently, leading to knowledge gaps within the current virus' situation. Nearly 20 years after the first evidence of West Nile virus in the archipelago, it has not yet been isolated, its impact on human and animal populations is unknown, and its local epidemiological cycle is still poorly understood. Within the framework of a pilot project started in Guadeloupe in 2019, West Nile virus was chosen as a federative model to apply the “One Health” approach for zoonotic epidemiological surveillance and shift from a sectorial to an integrated surveillance system. Human, animal, and environmental health actors involved in both research and surveillance were considered. Semi-directed interviews and a Social Network Analysis were carried out to learn about the surveillance network structure and actors, analyze information flows, and identify communication challenges. An information system was developed to fill major gaps: users' needs and main functionalities were defined through a participatory process where actors also tested and validated the tool. Additionally, all actors shared their data, which were digitized, cataloged, and centralized, to be analyzed later. An R Shiny server was integrated into the information system, allowing an accessible and dynamic display of data showcasing all of the partners' information. Finally, a series of virtual workshops were organized among actors to discuss preliminary results and plan the next steps to improve West Nile Virus and vector-borne or emerging zoonosis surveillance. The actors are willing to build a more resilient and cooperative network in Guadeloupe with improved relevance, efficiency, and effectiveness of their work

Forest biomass density across large climate gradients in northern South America is related to water availability but not with temperature

Understanding and predicting the likely response of ecosystems to climate change are crucial challenges for ecology and for conservation biology. Nowhere is this challenge greater than in the tropics as these forests store more than half the total atmospheric carbon stock in their biomass. Biomass is determined by the balance between biomass inputs (i.e., growth) and outputs (mortality). We can expect therefore that conditions that favor high growth rates, such as abundant water supply, warmth, and nutrient-rich soils will tend to correlate with high biomass stocks. Our main objective is to describe the patterns of above ground biomass (AGB) stocks across major tropical forests across climatic gradients in Northwestern South America. We gathered data from 200 plots across the region, at elevations ranging between 0 to 3400 m. We estimated AGB based on allometric equations and values for stem density, basal area, and wood density weighted by basal area at the plotlevel. We used two groups of climatic variables, namely mean annual temperature and actual evapotranspiration as surrogates of environmental energy, and annual precipitation, precipitation seasonality, and water availability as surrogates of water availability. We found that AGB is more closely related to water availability variables than to energy variables. In northwest South America, water availability influences carbon stocks principally by determining stand structure, i.e. basal area. When water deficits increase in tropical forests we can expect negative impact on biomass and hence carbon storage

A New High-Throughput Tool to Screen Mosquito-Borne Viruses in Zika Virus Endemic/Epidemic Areas

International audienceMosquitoes 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 BioMark™ 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