Host influence on the eukaryotic virome of sympatric mosquitoes and abundance of diverse viruses with a broad host range.

Mosquitoes harbor a large diversity of eukaryotic viruses. Those viromes probably influence mosquito physiology and the transmission of human pathogens. Nevertheless, their ecology remains largely unstudied. Here, we address two key questions in virome ecology. First, we assessed the influence of mosquito species on virome taxonomic diversity and relative abundance. Contrary to most previous studies, the potential effect of the habitat was explicitly included. Thousands of individuals of Culex poicilipes and Culex tritaeniorhynchus, two vectors of viral diseases, were concomitantly sampled in three habitats over two years. A total of 95 viral taxa from 25 families were identified with meta-transcriptomics, with 75% of taxa shared by both mosquitoes. Viromes significantly differed by mosquito species but not by habitat. Differences were largely due to changes in relative abundance of shared taxa. Then, we studied the diversity of viruses with a broad host range. We searched for viral taxa shared by the two Culex species and Aedes vexans, another disease vector, present in one of the habitats. Twenty-six out of the 163 viral taxa were found in the three mosquitoes. These taxa encompassed 14 families. A database analysis supported broad host ranges for many of those viruses, as well as a widespread geographical distribution. Thus, the viromes of mosquitoes from the same genera mainly differed in the relative abundance of shared taxa, whereas differences in viral diversity dominated between mosquito genera. Whether this new model of virome diversity and structure applies to other mosquito communities remains to be determined.

Mosquito (Diptera: Culicidae) populations in contrasting areas of the western regions of Burkina Faso: species diversity, abundance and their implications for pathogen transmission.

BACKGROUND: Mosquitoes (Diptera: Culicidae) can have a significant negative impact on human health. The urbanization of natural environments and their conversion for agricultural use, as well as human population growth, may affect mosquito populations and increase the risk of emerging or re-emerging mosquito-borne diseases. We report on the variety and number of adult mosquitoes found in four environments with varying degrees of human impact (rural, urban, rice fields, and forest) located in a savannah zone of West Africa. METHODS: Mosquitoes were collected from two regions (Hauts-Bassins and Sud-Ouest) of Burkina Faso during five periods between August 2019 and June 2021. Sampling sites were grouped according to environment. Mosquitoes were collected using BG-Sentinel traps and double net traps, and Prokopack Aspirators. Statistical analyses were performed using R software version 4.1.2. Logistic regression, using generalised mixed linear models, was used to test the effect of environment on mosquito abundance and diversity. Alpha diversity analysis was also performed, using the vegan package. RESULTS: A total of 10,625 adult mosquitoes were collected, belonging to 33 species and five genera: Culex, Aedes, Anopheles, Mansonia, and Ficalbia. The most dominant species were Culex quinquefasciatus, Anopheles gambiae sensu lato and Aedes aegypti. Alpha diversity was similar in the two regions. Habitat had a significant effect on mosquito species richness, the Shannon index and the Simpson index. The rural environment had the highest species richness (n = 28) followed by the forest environment (n = 24). The highest number of mosquitoes (4977/10,625) was collected in the urban environment. CONCLUSIONS: The species composition of the mosquito populations depended on the type of environment, with fewer species in environments with a high human impact such as urban areas and rice fields. Due to the diversity and abundance of the mosquito vectors, the human populations of all of the environments examined are considered to be at potential risk of mosquito-borne diseases.

Spatial scale influences the distribution of viral diversity in the eukaryotic virome of the mosquito Culex pipiens.

Our knowledge of the diversity of eukaryotic viruses has recently undergone a massive expansion. This diversity could influence host physiology through yet unknown phenomena of potential interest to the fields of health and food production. However, the assembly processes of this diversity remain elusive in the eukaryotic viromes of terrestrial animals. This situation hinders hypothesis-driven tests of virome influence on host physiology. Here, we compare taxonomic diversity between different spatial scales in the eukaryotic virome of the mosquito Culex pipiens. This mosquito is a vector of human pathogens worldwide. The experimental design involved sampling in five countries in Africa and Europe around the Mediterranean Sea and large mosquito numbers to ensure a thorough exploration of virus diversity. A group of viruses was found in all countries. This core group represented a relatively large and diverse fraction of the virome. However, certain core viruses were not shared by all host individuals in a given country, and their infection rates fluctuated between countries and years. Moreover, the distribution of coinfections in individual mosquitoes suggested random co-occurrence of those core viruses. Our results also suggested differences in viromes depending on geography, with viromes tending to cluster depending on the continent. Thus, our results unveil that the overlap in taxonomic diversity can decrease with spatial scale in the eukaryotic virome of C. pipiens. Furthermore, our results show that integrating contrasted spatial scales allows us to identify assembly patterns in the mosquito virome. Such patterns can guide future studies of virome influence on mosquito physiology.

Knowledge of Pelvic Floor Dysfunction in African American and Afro-Caribbean Women Seeking Medical Care in a Primary Care Ambulatory Setting.

IMPORTANCE: Approximately one fourth of U.S. community-dwelling women will develop a pelvic floor dysfunction (PFD) within their lifetimes. Prior research has revealed that knowledge of PFD was low to moderate in the general population and lower among Black patients. OBJECTIVE: This study aimed to assess the proficiency of urinary incontinence and pelvic organ prolapse (POP) in self-identified African American and Afro-Caribbean adult (age ≥ 18 years) female patients seeking medical care in our ambulatory setting. STUDY DESIGN: In this cross-sectional study, we administered the Prolapse and Incontinence Knowledge Questionnaire to patients in primary care and gynecology ambulatory settings at an academic medical center in Central Brooklyn. We used a multivariable Poisson regression model to find characteristics of the participants that are associated with proficiency in Prolapse and Incontinence Knowledge Questionnaire domains. RESULTS: A total of 266 survey participants self-identified as African American or Afro-Caribbean. Overall, using a multivariable model, knowledge of POP was significantly higher among African Americans than Afro-Caribbeans, and 75.5% of our patients reported that they would seek information on urinary incontinence and POP from a medical provider (gynecologist or primary care doctor) compared with other alternatives (eg, internet, 19.6%). CONCLUSIONS: These findings highlight subgroups that could benefit from provider-initiated education regarding PFD. Furthermore, although Black patients are often homogenized in research studies, differences may exist within subgroups likely because of varying interplays of structural racism and other social determinants of health, which may serve as an area of future research.

Screening of Circulation of Usutu and West Nile Viruses: A One Health Approach in Humans, Domestic Animals and Mosquitoes in Burkina Faso, West Africa.

Usutu virus (USUV) and West Nile virus (WNV) are phylogenetically closely related arboviruses. These viruses mainly follow an enzootic cycle involving mosquitoes and birds, but they occasionally infect humans and other mammals, inducing neurotropic disorders. Since the discovery of USUV, only two human cases have been reported in Africa, including one in Burkina Faso in 2004. Since then, no studies have been conducted to measure the extent of the circulation of this virus in Burkina Faso, and no study regarding the circulation of WNV has been conducted. Our study aimed to determine the seroprevalence of USUV and WNV in blood donations and in animals (horses, dogs, chickens and pigeons) and to perform molecular screening in patients with febrile fever and in Culex quinquefasciatus and Aedes aegypti mosquitoes. The prevalence of USUV and WNV was studied by serological (ELISA and microneutralization tests) and molecular analyses (RT-qPCR) of mosquito, dog, domestic bird, horse, and human samples in Burkina Faso between 2019 and 2021. We detected a very active transmission of both viruses in Burkina Faso. WNV and USUV seroprevalence is particularly high in humans (19.16% and 14.17%, respectively) and horses (17.28% and 6.17%). Molecular screening did not detect WNV or USUV in the mosquito or human samples tested. Our study shows an active spread of USUV and WNV in Burkina Faso, especially for WNV. This study highlights the value of developing surveillance programs to better prevent, detect, and alert people to USUV and WNV circulation in both primary and incidental hosts.

Serological Evidence of Zika Virus Circulation in Burkina Faso.

Zika virus (ZIKV) and dengue virus (DENV) are two closely related members of the Flaviviridae family, both transmitted by mosquitoes of the genus Aedes, and are among the arboviruses most at risk to human health. Burkina Faso has been facing an upsurge in DENV outbreaks since 2013. Unlike DENV, there is no serological evidence of ZIKV circulation in humans in Burkina Faso. The main objective of our study was to determine the seroprevalence of ZIKV and DENV in blood donors in Burkina Faso. A total of 501 donor samples collected in the two major cities of the country in 2020 were first tested by a competitive enzyme-linked immunosorbent assay to detect flavivirus antibodies. Positive sera were then tested using Luminex to detect ZIKV and DENV antibodies and virus-specific microneutralization tests against ZIKV were performed. The ZIKV seroprevalence was 22.75% in the donor samples and we found seropositivity for all DENV-serotypes ranging from 19.56% for DENV-1 to 48.86% for DENV-2. Molecular analyses performed on samples from febrile patients and Aedes aegypti mosquitoes between 2019 and 2021 were negative. Our study showed the important circulation of ZIKV and DENV detected by serology although molecular evidence of the circulation of ZIKV could not be demonstrated. It is essential to strengthen existing arbovirus surveillance in Burkina Faso and more broadly in West Africa by focusing on fevers of unknown origin and integrating vector surveillance to assess the extent of ZIKV circulation and identify the circulating strain. Further studies are needed to better understand the epidemiology of this virus in order to define appropriate prevention and response methods.

One Health surveillance of West Nile and Usutu viruses: a repeated cross-sectional study exploring seroprevalence and endemicity in Southern France, 2016 to 2020.

BackgroundWest Nile virus (WNV) and Usutu virus (USUV), two closely related flaviviruses, mainly follow an enzootic cycle involving mosquitoes and birds, but also infect humans and other mammals. Since 2010, their epidemiological situation may have shifted from irregular epidemics to endemicity in several European regions; this requires confirmation, as it could have implications for risk assessment and surveillance strategies.AimTo explore the seroprevalence in animals and humans and potential endemicity of WNV and USUV in Southern France, given a long history of WNV outbreaks and the only severe human USUV case in France in this region.MethodsWe evaluated the prevalence of WNV and USUV in a repeated cross-sectional study by serological and molecular analyses of human, dog, horse, bird and mosquito samples in the Camargue area, including the city of Montpellier, between 2016 and 2020.ResultsWe observed the active transmission of both viruses and higher USUV prevalence in humans, dogs, birds and mosquitoes, while WNV prevalence was higher in horses. In 500 human samples, 15 were positive for USUV and 6 for WNV. Genetic data showed that the same lineages, WNV lineage 1a and USUV lineage Africa 3, were found in mosquitoes in 2015, 2018 and 2020.ConclusionThese findings support existing literature suggesting endemisation in the study region and contribute to a better understanding of USUV and WNV circulation in Southern France. Our study underlines the importance of a One Health approach for the surveillance of these viruses.

Sensitivity and specificity analysis of 2D small field measurement array: Patient-specific quality assurance of small target treatments and spatially fractionated radiotherapy.

PURPOSE: The aim of this paper is to describe the tests carried out on a SRSMapCheck array, to verify its reliability and sensitivity for quality assurance (QA) of high gradient treatments as an alternative system to the use of high spatial resolution detectors, such as gafchromic film, whose processing requires meticulous and time-consuming procedures. METHODS: In an initial step, general functionality tests were carried out to verify that the equipment meets the manufacturer's specifications. A study of the accuracy of the application of correction factors to compensate for variation in detector response due to dose rate, field size and beam angle incidence has been included. Besides, to assess the ability of the array to detect inaccurately delivered treatments, systematic errors corresponding to the deviation in the position of the leaves and the accuracy of the gantry position, have been introduced. Based on these results, an estimate of sensitivity and specificity values of the device has been completed. The final step included a study applied to high gradient treatment for real cases of spatially fractionated radiotherapy, where the results of SRSMapCheck measurements have been compared with gafchromic films. RESULTS: General commissioning tests meet the manufacturer's specifications. dose rate (DR) response variation is better than 1.5% and for DR above 50 MU/min better than 1%. The results for beam incidences are better than 1% for all gantry angles, including beam incidences parallel to the array. Field size response differences are within the range of ±1% for sizes up to 2 × 2 cm2 , with a maximum value obtained of 3.5%, for 1 × 1 cm2 . From the systematic error study, using a Gamma function Γ (2%, 2 mm), the detector presents a high specificity with a value greater than 90% at its lower limit, while its sensitivity has a moderate mean value of 81%. Sensitivity values increase above 86% when we apply a Gamma function Γ (2%, 1 mm) is applied. Finally, the study of actual cases comprises 17 patients, distributed into 11 lung tumors, 3 gynecological and 3 soft tissue tumors. The gafchromic film showed a lower passing rate with an average value of Γ (2%, 2 mm) = 94.1% compared to Γ (2%, 2 mm) = 98.6% reached by the measurements with the array. CONCLUSIONS: Gamma function obtained with the SRSMapCheck array always presented a higher value than gafchromic film measurements, resulting in a greater number of plans considered correct. This fact, together with the sensitivity and specificity study carried out, allows us to conclude the recommendation that a restrictive metric must be established, in this way we will improve sensitivity, and therefore we will reduce the rate of incorrect plans qualified as correct. The characteristics of the equipment together with the correction factors applied, led to reliably performing acquisitions for complex treatments with multiple small targets in oblique rotational incidences. The spatial resolution of detectors allows the verification of high gradient dose plans such as those achieved in spatially fractionated radiotherapy (SFRT).

A library preparation optimized for metagenomics of RNA viruses.

Our understanding of the viral communities associated to animals has not yet reached the level attained on the bacteriome. This situation is due to, among others, technical challenges in adapting metagenomics using high-throughput sequencing to the study of RNA viromes in animals. Although important developments have been achieved in most steps of viral metagenomics, there is yet a key step that has received little attention: the library preparation. This situation differs from bacteriome studies in which developments in library preparation have largely contributed to the democratisation of metagenomics. Here, we present a library preparation optimized for metagenomics of RNA viruses from insect vectors of viral diseases. The library design allows a simple PCR-based preparation, such as those routinely used in bacterial metabarcoding, that is adapted to shotgun sequencing as required in viral metagenomics. We first optimized our library preparation using mock viral communities and then validated a full metagenomic approach incorporating our preparation in two pilot studies with field-caught insect vectors; one including a comparison with a published metagenomic protocol. Our approach provided a fold increase in virus-like sequences compared to other studies, and nearly-full genomes from new virus species. Moreover, our results suggested conserved trends in virome composition within a population of a mosquito species. Finally, the sensitivity of our approach was compared to a commercial diagnostic PCR for the detection of an arbovirus in field-caught insect vectors. Our approach could facilitate studies on viral communities from animals and the democratization of metagenomics in community ecology of viruses.

Driver glare exposure with different vehicle frontlighting systems.

INTRODUCTION: Highway safety performance at night has received less attention in research than daytime, despite the higher accident rates occurring under night-time conditions. This study presents a procedure to assess the potential hazard for drivers created by headlight glare and its interaction with the geometric design of highways. METHOD: The proposed procedure consists of a line-of-sight analysis performed by a geoprocessing model in geographic information systems to determine whether the rays of light that connect headlights and oncoming drivers are obstructed by either the roadway or its roadsides. Then, the procedure checks whether the non-obstructed rays of light are enclosed by a given headlight beam. Different hypotheses were set concerning the headlight beam features, including the horizontal spread angle and whether the headlights are fixed or swiveling. A highway section was selected to test and validate the procedure proposed. A 3D recreation of the highway and its environment derived from a LiDAR point cloud was used for this purpose. RESULTS: The findings disclose how glare is produced on tangents, horizontal curves, transitions between them and sequences of curves. The effect of visual obstructions conveniently placed is also discussed. CONCLUSIONS: A greater glare incidence is produced as the horizontal headlights spread angle increases. Swiveling headlights increase glare on highways left curves and reduce it on right curves. Practical Applications: The procedure and conclusions of this study can contribute to develop more effective glare avoidance technologies as well as identify and assess glare-prone sections. The glare evaluation assists in evaluating glare countermeasures such as deciding whether to place a vegetation barrier and where.

Identification of Umbre Orthobunyavirus as a Novel Zoonotic Virus Responsible for Lethal Encephalitis in 2 French Patients with Hypogammaglobulinemia.

BACKGROUND: Human encephalitis represents a medical challenge from a diagnostic and therapeutic point of view. We investigated the cause of 2 fatal cases of encephalitis of unknown origin in immunocompromised patients. METHODS: Untargeted metatranscriptomics was applied on the brain tissue of 2 patients to search for pathogens (viruses, bacteria, fungi, or protozoans) without a prior hypothesis. RESULTS: Umbre arbovirus, an orthobunyavirus never previously identified in humans, was found in 2 patients. In situ hybridization and reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) showed that Umbre virus infected neurons and replicated at high titers. The virus was not detected in cerebrospinal fluid by RT-qPCR. Viral sequences related to Koongol virus, another orthobunyavirus close to Umbre virus, were found in Culex pipiens mosquitoes captured in the south of France where the patients had spent some time before the onset of symptoms, demonstrating the presence of the same clade of arboviruses in Europe and their potential public health impact. A serological survey conducted in the same area did not identify individuals positive for Umbre virus. The absence of seropositivity in the population may not reflect the actual risk of disease transmission in immunocompromised individuals. CONCLUSIONS: Umbre arbovirus can cause encephalitis in immunocompromised humans and is present in Europe.

The Genome Segments of Bluetongue Virus Differ in Copy Number in a Host-Specific Manner.

Genome segmentation is mainly thought to facilitate reassortment. Here, we show that segmentation can also allow differences in segment abundance in populations of bluetongue virus (BTV). BTV has a genome consisting in 10 segments, and its cycle primarily involves periodic alternation between ruminants and Culicoides biting midges. We have developed a reverse transcription-quantitative PCR (RT-qPCR) approach to quantify each segment in wild BTV populations sampled in both ruminants and midges during an epizootic. Segment frequencies deviated from equimolarity in all hosts. Interestingly, segment frequencies were reproducible and distinct between ruminants and biting midges. Beyond a putative regulatory role in virus expression, this phenomenon could lead to different evolution rates between segments.IMPORTANCE The variation in viral gene frequencies remains a largely unexplored aspect of within-host genetics. This phenomenon is often considered to be specific to multipartite viruses. Multipartite viruses have segmented genomes, but in contrast to segmented viruses, their segments are each encapsidated alone in a virion. A main hypothesis explaining the evolution of multipartism is that, compared to segmented viruses, it facilitates the regulation of segment abundancy, and the genes the segments carry, within a host. These differences in gene frequencies could allow for expression regulation. Here, we show that wild populations of a segmented virus, bluetongue virus (BTV), also present unequal segment frequencies. BTV cycles between ruminants and Culicoides biting midges. As expected from a role in expression regulation, segment frequencies tended to show specific values that differed between ruminants and midges. Our results expand previous knowledge on gene frequency variation and call for studies on its role and conservation beyond multipartite viruses.

Updated unified phylogenetic classification system and revised nomenclature for Newcastle disease virus.

Several Avian paramyxoviruses 1 (synonymous with Newcastle disease virus or NDV, used hereafter) classification systems have been proposed for strain identification and differentiation. These systems pioneered classification efforts; however, they were based on different approaches and lacked objective criteria for the differentiation of isolates. These differences have created discrepancies among systems, rendering discussions and comparisons across studies difficult. Although a system that used objective classification criteria was proposed by Diel and co-workers in 2012, the ample worldwide circulation and constant evolution of NDV, and utilization of only some of the criteria, led to identical naming and/or incorrect assigning of new sub/genotypes. To address these issues, an international consortium of experts was convened to undertake in-depth analyses of NDV genetic diversity. This consortium generated curated, up-to-date, complete fusion gene class I and class II datasets of all known NDV for public use, performed comprehensive phylogenetic neighbor-Joining, maximum-likelihood, Bayesian and nucleotide distance analyses, and compared these inference methods. An updated NDV classification and nomenclature system that incorporates phylogenetic topology, genetic distances, branch support, and epidemiological independence was developed. This new consensus system maintains two NDV classes and existing genotypes, identifies three new class II genotypes, and reduces the number of sub-genotypes. In order to track the ancestry of viruses, a dichotomous naming system for designating sub-genotypes was introduced. In addition, a pilot dataset and sub-trees rooting guidelines for rapid preliminary genotype identification of new isolates are provided. Guidelines for sequence dataset curation and phylogenetic inference, and a detailed comparison between the updated and previous systems are included. To increase the speed of phylogenetic inference and ensure consistency between laboratories, detailed guidelines for the use of a supercomputer are also provided. The proposed unified classification system will facilitate future studies of NDV evolution and epidemiology, and comparison of results obtained across the world.

Identification of Eilat virus and prevalence of infection among Culex pipiens L. populations, Morocco, 2016.

Eilat virus (EILV) is described as one of the few alphaviruses restricted to insects. We report the record of a nearly-complete sequence of an alphavirus genome showing 95% identity with EILV during a metagenomic analysis performed on 1488 unblood-fed females and 1076 larvae of the mosquito Culex pipiens captured in Rabat (Morocco). Genetic distance and phylogenetic analyses placed the EILV-Morocco as a variant of EILV. The observed infection rates in both larvae and adults suggested an active circulation of the virus in Rabat and its maintenance in the environment either through vertical transmission or through horizontal infection of larvae in breeding sites. This is the first report of EILV out of Israel and in Culex pipiens populations.

Sobrepeso/obesidad en niños en edad escolar y sus factores de riesgo

Introducción: sobrepeso y obesidad son un problema de salud pública, con una prevalencia creciente en niños uruguayos. El diagnóstico precoz y la identificación de factores de riesgo son fundamentales para su tratamiento y prevención. Objetivos: evaluar la prevalencia de sobrepeso/obesidad en niños escolares e identificar factores de riesgo. Metodología: estudio transversal, realizado entre julio de 2015 y julio de 2016 a niños de 4° y 5° año de escuelas públicas y privadas de Rivera y Montevideo. Se recogieron variables patronímicas, peso al nacer, datos sobre su alimentación en el primer año de vida y datos antropométricos. Se interrogó frecuencia de consumo de algunos alimentos y nivel de actividad física. Se describió la prevalencia de sobrepeso/obesidad. Se compararon factores de riesgo, consumo de alimentos y actividad física entre niños con y sin sobrepeso/obesidad. Resultados: se incluyeron 318 niños; 28,3% tenía sobrepeso, 14,5% obesidad. Los niños con y sin sobrepeso/obesidad no mostraron diferencias significativas con respecto al sexo, pertenecer a instituciones públicas, tener bajo peso al nacer, o la alimentación recibida en el primer año de vida. Mayor proporción de niños con sobrepeso/obesidad recibía galletitas, bizcochos, snacks, golosinas y dulces, y permanecía más de 6 horas sentados; menor proporción tenía más de 60 minutos/día de juegos al aire libre (p<0,05). Conclusiones: la prevalencia de sobrepeso/obesidad en el grupo evaluado fue elevada. El consumo de alimentos hipercalóricos y la menor actividad física se asociaron significativamente a su desarrollo. La etapa escolar es un momento crucial en la modificación de hábitos que posibilita la prevención del trastorno.

Summary: Introduction: overweight and obesity are a public health problem, with an increasing prevalence in Uruguayan children. Early diagnosis and identification of risk factors are essential for treatment and prevention. Objective: to evaluate the prevalence of overweight and obesity in school-aged children and to identify risk factors. Methodology: cross-sectional study, carried out between July, 2015 and July, 2016 on 4th and 5th grade children of public and private schools of Rivera and Montevideo. Patronymic variables, birth weight, first year of life feeding data and anthropometric data were collected. Frequency of consumption of some foods and level of physical activity was studied. The prevalence of overweight and obesity was described. Risk factors, food consumption and physical activity among overweight and obese children were compared. Results: a total of 318 children were included; 28.3% were overweight, 14.5% were obese. Children with and without overweight/obesity did not show significant differences regarding sex, belonging to public institutions, having low birth weight, or considering the food they consumed in the first year of life. A higher proportion of overweight/obese children received cookies, biscuits, snacks, and sweets, and remained more than 6 hours sitting; a lower proportion had more than 60 minutes/day of outdoor games (p <0.05). Conclusions: the prevalence of overweight and obesity in the evaluated group was high. The consumption of hyper caloric foods and the lower physical activity were significantly associated to its development. The school stage is a crucial moment in the modification of habits that allow the prevention of the disorder.

Introdução: o sobrepeso e a obesidade são um problema de saúde pública com prevalência crescente em crianças uruguaias. O diagnóstico precoce e a identificação de fatores de risco são fundamentais para o seu tratamento e prevenção. Objetivos: avaliar a prevalência de sobrepeso / obesidade em escolares e identificar fatores de risco. Metodologia: estudo transversal, realizado entre julho / 2015 e julho / 2016 para crianças com 4 e 5 anos de idade de escolas públicas e privadas nas cidades de Rivera e Montevidéu. Foram coletadas variáveis patronímicas, peso ao nascer, dados sobre sua dieta no primeiro ano de vida e dados antropométricos. A pesquisa perguntou sobre a frequência de consumo de alguns alimentos e o nível de atividade física. Descrevemos a prevalência de sobrepeso / obesidade e comparamos fatores de risco, consumo de alimentos e atividade física entre crianças com e sem sobrepeso / obesidade. Resultados: 318 crianças participaram do estudo; 28,3% tinham sobrepeso, 14,5% eram obesos. As crianças com e sem sobrepeso / obesidade não apresentaram diferenças significativas em relação ao sexo, por frequentar escolas públicas, com baixo peso ao nascer ou por causa dos alimentos recebidos no primeiro ano de vida. A maior proporção de crianças com sobrepeso / obesidade consume biscoitos, bolachas, salgadinhos, doces e guloseimas e permanece por mais de 6 horas sentada; a menor proporção passou mais de 60 minutos / dia de jogos ao ar livre (p <0,05). Conclusões: a prevalência de sobrepeso / obesidade no grupo avaliado foi alta. O consumo de alimentos hipercalóricos e a menor atividade física foram significativamente associados ao seu desenvolvimento. A etapa de educação escolar é um momento crucial na modificação de hábitos que possibilitem a prevenção do transtorno.

Can genotype mismatch really affect the level of protection conferred by Newcastle disease vaccines against heterologous virulent strains?

Newcastle disease (ND), caused by virulent class II avian paramyxovirus 1 (Newcastle disease virus, NDV), occurs sporadically in poultry despite their having been immunized with commercial vaccines. These vaccines were all derived from NDV strains isolated around 70 years ago. Since then, class II NDV strains have evolved into 18 genotypes. Whether the vaccination failure results from genotype mismatches between the currently used vaccine strains and field-circulating velogenic strains or from an impaired immune response in the vaccination remains unclear. To test the first hypothesis, we performed a heterologous genotype II vaccine/genotype XI challenge in one-day old specific pathogen free (SPF) chicks and reproduced viral shedding. We then produced two attenuated strains of genotype II and XI by reverse genetics and used them to immunize two-week old SPF chickens that were subsequently challenged with velogenic strains of genotypes II, VII and XI. We found that both vaccines could induce antibodies with hemagglutination inhibition titers higher than 6.5 log2. Vaccination also completely prevented disease, viral shedding in swabs, and blocked viral replication in tissues from different genotypes in contrast to unvaccinated chickens that died shortly after challenge. Taken together, our results support the hypothesis that, in immunocompetent poultry, genotype mismatch is not the main reason for vaccination failure.

Human Usutu Virus Infection with Atypical Neurologic Presentation, Montpellier, France, 2016.

Infection with Usutu virus (USUV) has been recently associated with neurologic disorders, such as encephalitis or meningoencephalitis, in humans. These findings indicate that USUV is a potential health threat. We report an acute human infection with USUV in France putatively associated with a clinical diagnosis of idiopathic facial paralysis.

Emergence of two Usutu virus lineages in Culex pipiens mosquitoes in the Camargue, France, 2015.

Usutu virus (USUV) was detected in 11 Culex pipiens mosquito pools collected in 2015 in Camargue (France) using quantitative real-time RT-PCR assays. Phylogenetic analysis of recovered virus sequences identified USUV lineages Africa 2 and Africa 3, demonstrating the simultaneous occurrence of different strains within the mosquito population. This is the first report on USUV in mosquitoes from France that concurrently accompanied the emergence of Usutu virus in blackbirds and a human case in France during 2015/2016.

Cleavage site of Newcastle disease virus determines viral fitness in persistent infection cells.

Newcastle disease, caused by infection with virulent strains of Newcastle disease virus (NDV), poses a risk for the poultry industry. The virulence of NDV is mainly determined by the cleavage site of F protein. Lentogenic NDV can become velogenic after passages in SPF chicken brain and air sac based on some strains isolated from water birds, because the proportion of virulent-related strains gradually increases. In contrast, this proportion remains unchanged if NDV is passaged via 10-day-old SPF chicken embryos. This information suggests that environmental conditions rather than mutation affect NDV fitness in quasispecies. However, it is unknown how the environment selects virulent-related strains from a viral population. In this study, velogenic and lentogenic NDV marked by green or red fluorescence were used to establish persistent infection (PI) in BHK-21 cells. Monitoring viruses by different methods, we found that, without competition, persistently infected cells harbored lentogenic and velogenic NDV strains similarly in terms of viral release, viral spread and the period of persistent viral infection. In contrast, under competitive co-infection, velogenic NDV became dominant in quasispecies from the fifth passage of PI cells, which resulted in the progressive disappearance of the lentogenic NDV strain. This domination was concomitant with a short-term reduction in the superinfection exclusion and supernatant interference in PI cells resulting in a velogenic virus rebound. We concluded that virulent-related F protein cleavage site facilitates the spread and replication of NDV in conditions under which cells do not secret trypsin-like proteases and do not inhibit free virus infection, resulting in a gradual increase in virulent strains in quasispecies with the number of passages.

Comparison of the efficiency of different newcastle disease virus reverse genetics systems.

Rescue of negative-sense single-stranded RNA viruses ((-)ssRNA virus), generally requires the handling of a large number of plasmids to provide the virus genome and essential components for gene expression and genome replication. This constraint probably renders reverse genetics of (-)ssRNA virus more complex and less efficient. Some authors have shown that the fewer the plasmids, the more efficient reverse genetics is for segmented RNA virus. However, it is not clear if the same applies for (-)ssRNA, such as Newcastle disease virus (NDV). To address this issue, six variants of NDV reverse genetic systems were established by cloning combinations of NP, P and L genes, mini-genome or full-genome in 4, 3, 2 and 1 plasmid. In terms of mini-genome and full-genome rescue, we showed that only the 2-plasmid system, assembling three support plasmids together, was able to improve the rescue efficiency over that of the conventional 4-plasmid system. These results may help establish and/or improve reverse genetics for other mononegaviruses.