Shared larval rearing environment, sex, female size and genetic diversity shape Ae. albopictus bacterial microbiota

The Asian tiger mosquito Aedes albopictus became of public health concern as it can replicate and transmit viral and filarial pathogens with a strong invasive success over the world. Various strategies have been proposed to reduce mosquito population's vectorial capacity. Among them, symbiotic control of mosquito borne disease offers promising perspectives. Such method is likely to be affected by the dynamics of mosquito-associated symbiotic communities, which might in turn be affected by host genotype and environment. Our previous study suggested a correlation between mosquitoes' origin, genetic diversity and midgut bacterial diversity. To distinguish the impact of those factors, we have been studying the midgut bacterial microbiota of two Ae. albopictus populations from tropical (La Re A union) and temperate (Montpellier) origins under controlled laboratory conditions. the two populations experienced random mating or genetic bottleneck. Microbiota composition did not highlight any variation of the alpha and beta-diversities in bacterial communities related to host's populations. However, sizes of the mosquitoes were negatively correlated with the bacterial a-diversity of females. Variations in mosquito sex were associated with a shift in the composition of bacterial microbiota. The females' mosquitoes also exhibited changes in the microbiota composition according to their size and after experiencing a reduction of their genetic diversity. These results provide a framework to investigate the impact of population dynamics on the symbiotic communities associated with the tiger mosquito.Peer reviewe

Extracorporeal Membrane Oxygenation for Severe Acute Respiratory Distress Syndrome associated with COVID-19: An Emulated Target Trial Analysis.

RATIONALE: Whether COVID patients may benefit from extracorporeal membrane oxygenation (ECMO) compared with conventional invasive mechanical ventilation (IMV) remains unknown. OBJECTIVES: To estimate the effect of ECMO on 90-Day mortality vs IMV only Methods: Among 4,244 critically ill adult patients with COVID-19 included in a multicenter cohort study, we emulated a target trial comparing the treatment strategies of initiating ECMO vs. no ECMO within 7 days of IMV in patients with severe acute respiratory distress syndrome (PaO2/FiO2 <80 or PaCO2 ≥60 mmHg). We controlled for confounding using a multivariable Cox model based on predefined variables. MAIN RESULTS: 1,235 patients met the full eligibility criteria for the emulated trial, among whom 164 patients initiated ECMO. The ECMO strategy had a higher survival probability at Day-7 from the onset of eligibility criteria (87% vs 83%, risk difference: 4%, 95% CI 0;9%) which decreased during follow-up (survival at Day-90: 63% vs 65%, risk difference: -2%, 95% CI -10;5%). However, ECMO was associated with higher survival when performed in high-volume ECMO centers or in regions where a specific ECMO network organization was set up to handle high demand, and when initiated within the first 4 days of MV and in profoundly hypoxemic patients. CONCLUSIONS: In an emulated trial based on a nationwide COVID-19 cohort, we found differential survival over time of an ECMO compared with a no-ECMO strategy. However, ECMO was consistently associated with better outcomes when performed in high-volume centers and in regions with ECMO capacities specifically organized to handle high demand. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Diversity of culturable bacteria including Pantoea in wild mosquito Aedes albopictus

Background: The microbiota has been shown to play an important role in the biology of insects. In recent decades, significant efforts have been made to better understand the diversity of symbiotic bacteria associated with mosquitoes and assess their influence on pathogen transmission. Here, we report the bacterial composition found in field-caught Aedes albopictus populations by using culture-dependent methods. Results: A total of 104 mosquito imagos (56 males and 48 females) were caught from four contrasting biotopes of Madagascar and their bacterial contents were screened by plating whole body homogenates on three different culture media. From 281 bacterial colony types obtained, amplified ribosomal DNA restriction analysis (ARDRA) showed they had 40 distinct ribotypes. Sequencing and BLAST analysis of the 16S rDNA genes responsible for each representative profile made it possible to identify 27 genera distributed in three major phyla. In female mosquitoes, bacterial isolates were mostly Proteobacteria (51.3%) followed by Firmicutes (30.3%) and Actinobacteria (18.4%). Conversely, Actinobacteria was the most abundant phylum in male mosquitoes (48%) followed by Proteobacteria (30.6%) and Firmicutes (20.4%). The relative abundance and composition of isolates also varied between sampling sites, ranging from 3 distinct families in Ankazobe to 8 in Tsimbazaza Park, and Toamasina and Ambohidratrimo. Pantoea was the most common genus in both females and males from all sampling sites, except for Ambohidratrimo. No differences in genome size were found between Pantoea isolates from mosquitoes and reference strains in pulse field gel electrophoresis. However, according to the numbers and sizes of plasmids, mosquito isolates clustered into three different groups with other strains isolated from insects but distinct from isolates from the environment. Conclusions: The recent upsurge in research into the functional role of the insect microbiota prompts the interest to better explore the role some bacteria detected here may have in the mosquito biology. Future studies of culturable bacteria might decipher whether they have a biological role in the invasiveness of Ae. albopictus. As a possible candidate for paratransgenesis, the predominant genus Pantoea will be characterized to better understand its genetic contents and any possible influence it may have on vector competence of Ae. albopictus.Arthropod Symbioses: from fundamental to pest disease managemen

Short-term impacts of anthropogenic stressors on Aedes albopictus mosquito vector microbiota

Recent studies have highlighted the potential role of microbiota in the biology of the Aedes albopictus mosquito vector. This species is highly anthropogenic and exhibits marked ecological plasticity, with a resulting high potential to colonize a wide range of habitats-including anthropized areas-under various climatic conditions. We put forward the hypothesis that climate and anthropogenic activities, such as the use of antibiotics in agriculture and human medicine, might affect the mosquito-associated bacterial community. We thus studied the additive impact of a temperature decrease and antibiotic ingestion on the temporal dynamics of Ae. albopictus survival and its associated bacterial communities. The results showed no effects of disturbances on mosquito survival. However, short-term temperature impacts on bacterial diversity were observed, while both the community structure and bacterial diversity were affected by early antibiotic ingestion. The genera Elizabethkingia, Chryseobacterium and Wolbachia, as well as an unclassified member of the Bacteroidales order were particularly affected. Antibiotics negatively impacted Elizabethkingia abundance, while Chryseobacterium was completely eliminated following both disturbances, to the benefit of Wolbachia and the unclassified Bacteroidales species. These results generated fresh insight into the effects of climate and anthropogenic activities such as the use of antibiotics on mosquito microbiota

Are disulphide bonds formed during acid gelation of preheated milk?

The role of thiol/disulphide exchanges during acid gelation of preheated milk was studied with milk samples with or without N-ethylmaleimide (NEM), a thiol-blocking agent, and acidified to pH 4 by the addition of glucono-delta-lactone at 20 °C. Active or total thiol groups, particle size with light scattering measurements in a dissociating solvent or by SDS-agarose electrophoresis were determined on acidified milk samples. Diffusing wave spectroscopy and rheology in low strain were applied during acidification of sample, while rheology in large strain was applied on final acid gels. The only effect of the presence of NEM was a reduced firmness of acid gels as measured at large strain and a reduced tendency to form large aggregates at pH<5.5. In conclusions, thiol/disulphide exchanges during acidification of milk played only a minor role in the building of acid gel networks from heated milk

Aedes albopictus mosquitoes host a locally structured mycobiota with evidence of reduced fungal diversity in invasive populations

The Asian tiger mosquito Aedes albopictus, native to Southeast Asia, has invaded a wide range of tropical and temperate areas worldwide. Recent studies pointed out that invasive populations from Europe harbored reduced bacterial microbiota compared to the native populations. Beside bacteria, mosquitoes also contain fungal communities that have so far been largely ignored. To investigate whether the mosquito invasion process displays a similar impact on fungal diversity, we compared the mycobiota structure of three autochthonous mosquito populations in Vietnam and six populations recently introduced in France and Madagascar. All mosquito populations host a locally structured fungal community and carry a "core mycobiota" dominated by yeasts. However, invasive populations from France and Madagascar harbor a lower fungal diversity compared to Vietnamese populations. These results suggest that similar factors shape the overall composition of the mosquito-associated microbiota during the invasion process as bacterial and fungal communities demonstrate a loss of diversity

Native Wolbachia from Aedes albopictus Blocks Chikungunya Virus Infection In Cellulo

International audienceWolbachia, a widespread endosymbiont of terrestrial arthropods, can protect its host against viral and parasitic infections, a phenotype called "pathogen blocking". However, in some cases Wolbachia may have no effect or even enhance pathogen infection, depending on the host-Wolbachia-pathogen combination. The tiger mosquito Aedes albopictus is naturally infected by two strains of Wolbachia, wAlbA and wAlbB, and is a competent vector for different arboviruses such as dengue virus (DENV) and Chikungunya virus (CHIKV). Interestingly, it was shown in some cases that Ae. albopictus native Wolbachia strains are able to inhibit DENV transmission by limiting viral replication in salivary glands, but no such impact was measured on CHIKV replication in vivo. To better understand the Wolbachia/CHIKV/Ae. albopictus interaction, we generated a cellular model using Ae. albopictus derived C6/36 cells that we infected with the wAlbB strain. Our results indicate that CHIKV infection is negatively impacted at both RNA replication and virus assembly/secretion steps in presence of wAlbB. Using FISH, we observed CHIKV and wAlbB in the same mosquito cells, indicating that the virus is still able to enter the cell in the presence of the bacterium. Further work is needed to decipher molecular pathways involved in Wolbachia-CHIKV interaction at the cellular level, but this cellular model can be a useful tool to study the mechanism behind virus blocking phenotype induced by Wolbachia. More broadly, this underlines that despite Wolbachia antiviral potential other complex interactions occur in vivo to determine mosquito vector competence in Ae. albopictus

Potential role of autophagy during Wolbachia antiviral interference against chikungunya virus in mosquito cells

International audienceArthropod-borne viruses (Arboviruses) like dengue (DENV) or chikungunya (CHIKV) virus are a major threat for public health, responsible for millions of cases each year worldwide. Arboviruses are mainly transmitted by mosquitoes, whose vectorial competence depends on environmental, genetic and immune factors. Recently, it was showed that mosquito microflora, like fungi and bacteria, could also modulate host vectorial competence. This is the case for Wolbachia, an obligatory intracellular Alphaproteobacteria commonly found in arthropods, which was shown to decrease DENV, WNV and CHIKV transmission in mosquito. Nevertheless, the cellular mechanisms of this interaction remain largely unknown. To improve our understanding of the host mechanisms involved in Wolbachia-arbovirus interaction, we used the C6/36 cell line from Aedes albopictus, a competent vector of CHIKV and DENV. We stably infected C6/36 cells with the Wolbachia strain wAlbB, naturally present in Ae. albopictus, then performed CHIKV infections. Results showed that viral replication and infectiosity decrease in the presence of the bacteria compare to cells were Wolbachia was cleared using antibiotic. In light of recent studies suggesting the importance of autophagic process during arbovirus infection, we showed that autophagy was modulated in mosquito during viral and bacterial co-infection, and could partially explain the antiviral effect of Wolbachia. These results must be thorough and extended to other mosquito species to try to decipher the role of mosquito endosymbionts on pathogen transmission