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Despite the efforts made in recent decades, glioblastoma is still the deadliest primary brain cancer without cure. The potential role in tumour maintenance and progression of the peritumoural brain zone (PBZ), the apparently normal area surrounding the tumour, has emerged. Little is known about this area due to a lack of common definition and due to difficult sampling related to the functional role of peritumoural healthy brain. The aim of this work was to better characterize the PBZ and to identify genes that may have role in its malignant transformation. Starting from our previous study on the comparison of the genomic profiles of matched tumour core and PBZ biopsies, we selected CDK4 and EXT2 as putative malignant drivers of PBZ. The gene expression analysis confirmed their over-expression in PBZ, similarly to what happens in low-grade glioma and glioblastoma, and CDK4 high levels seem to negatively influence patient overall survival. The prognostic role of CDK4 and EXT2 was further confirmed by analysing the TCGA cohort and bioinformatics prediction on their gene networks and protein–protein interactions. These preliminary data constitute a good premise for future investigations on the possible role of CDK4 and EXT2 in the malignant transformation of PBZ.

During cancer treatments in childhood hematological malignancies, reduced exercise tolerance is one of the main hardships. Precision-based training programs help children, adolescents, and young adults and their families to resume regular physical activity, exercise, and sports once they return to their communities after the intensive phases spent in hospital. This study was aimed at verifying whether an intermittent recovery test, the Yo-Yo AD, could provide a simple and valid way to evaluate an individual’s capacity to perform repeated intense exercise and to follow up on the impact of tailored exercise in children, adolescents, and young adults with hematological malignancies. The Yo-Yo AD involved the repetition of several shuttles to muscle exhaustion, at pre-established speeds (walking and slow running). The heart rate (HR) and oxygen saturation (SaO2 ) were monitored during the test. The total distance and the walking/running ability, measured as the slope of the HR vs. distance correlation, were investigated before (T0) and after 11 weeks (T1) of precision exercise intervention. The Yo-Yo AD was also performed by healthy children (CTRL). Ninety-seven patients (10.58 ± 4.5 years, 46% female) were enrolled. The Yo-Yo AD showed the positive impact of the exercise intervention by increasing the distance covered by the individuals (T0 = 946.6 ± 438.2 vs. T1 = 1352.3 ± 600.6 m, p < 0.001) with a more efficient walking/running ability (T0 = 2.17 ± 0.84 vs. T1 = 1.73 ± 0.89 slope, p < 0.0164). CTRLs performed better (1754.0 ± 444.0 m, p = 0.010). They were equally skillful (1.71 ± 0.27 slope) when compared to the patients after they received the precision-based intervention. No adverse events occurred during the Yo-Yo AD and it proved to be an accurate way of correctly depicting the changes in performance in childhood hematological malignancies.

Our study investigated the effects of hypoxia on visuospatial attention processing during preparation for a single/double-choice motor response. ERPs were recorded in two sessions in which participants breathed either ambient-air or oxygen-impoverished air. During each session, participants performed four cue-target attention orienting and/or alerting tasks. Replicating the classic findings of valid visuospatial attentional orienting modulation, ERPs to pre-target cues elicited both an Anterior directing attention negativity (ADAN)/CNV and a posterior Late directing attention positivity (LDAP)/TP, which in ambient air were larger for attention orienting than for alerting. Hypoxia increased the amplitude of both these potentials in the spatial orienting conditions for the upper visual hemifield, while, for the lower hemifield, it increased ADAN/CNV, but decreased LDAP/TP for the same attention conditions. To these ERP changes corresponded compensatory enhanced activation of right anterior cingulate cortex, left superior parietal lobule and frontal gyrus, as well as detrimental effects of hypoxia on behavioral overt performance. Together, these findings reveal for the first time, to our knowledge, that (1) these reversed alterations of the activation patterns during the time between cue and target occur at a larger extent in hypoxia than in air, and (2) acute normobaric hypoxia alters visuospatial attention orienting shifting in space.

Two by-products containing phenols and polysaccharides, a “pâté” (OP) from the extra virgin olive oil milling process and a decoction of pomegranate mesocarp (PM), were investigated for their effects on human microbiota using the SHIME® system. The ability of these products to modulate the microbial community was studied simulating a daily intake for nine days. Microbial functionality, investigated in terms of short chain fatty acids (SCFA) and NH4+, was stable during the treatment. A significant increase in Lactobacillaceae and Bifidobacteriaceae at nine days was induced by OP mainly in the proximal tract. Polyphenol metabolism indicated the formation of tyrosol from OP mainly in the distal tract, while urolithins C and A were produced from PM, identifying the human donor as a metabotype A. The results confirm the SHIME® system as a suitable in vitro tool to preliminarily investigate interactions between complex botanicals and human microbiota before undertaking more challenging human studies.

Introduction: Parkinson's disease (PD) is one of the most frequent causes of disability among older people, characterized by motor disorders, rigidity, and balance problems. Recently, dance has started to be considered an effective exercise for people with PD. In particular, Irish dancing, along with tango and different forms of modern dance, may be a valid strategy to motivate people with PD to perform physical activity. The present protocol aims to implement and evaluate a rehabilitation program based on a new system called “SI-ROBOTICS,” composed of multiple technological components, such as a social robotic platform embedded with an artificial vision setting, a dance-based game, environmental and wearable sensors, and an advanced AI reasoner module. Methods and Analysis: For this study, 20 patients with PD will be recruited. Sixteen therapy sessions of 50 min will be conducted (two training sessions per week, for 8 weeks), involving two patients at a time. Evaluation will be primarily focused on the acceptability of the SI-ROBOTICS system. Moreover, the analysis of the impact on the patients' functional status, gait, balance, fear of falling, cardio-respiratory performance, motor symptoms related to PD, and quality of life, will be considered as secondary outcomes. The trial will start in November 2021 and is expected to end by April 2022. Discussions: The study aims to propose and evaluate a new approach in PD rehabilitation, focused on the use of Irish dancing, together with a new technological system focused on helping the patient perform the dance steps and on collecting kinematic and performance parameters used both by the physiotherapist (for the evaluation and planning of the subsequent sessions) and by the system (to outline the levels of difficulty of the exercise). Ethics and Dissemination: The study was approved by the Ethics Committee of the IRCCS INRCA. It was recorded in ClinicalTrials.gov on the number NCT05005208. The study findings will be used

Glioblastoma is an extremely heterogeneous disease. Treatment failure and tumor recurrence primarily reflect the presence in the tumor core (TC) of the glioma stem cells (GSCs), and secondly the contribution, still to be defined, of the peritumoral brain zone (PBZ). Using the array-CGH platform, we deepened the genomic knowledge about the different components of GBM and we identified new specific biomarkers useful for new therapies. We firstly investigated the genomic profile of 20 TCs of GBM; then, for 14 cases and 7 cases, respectively, we compared these genomic profiles with those of the related GSC cultures and PBZ biopsies. The analysis on 20 TCs confirmed the intertumoral heterogeneity and a high percentage of copy number alterations (CNAs) in GBM canonical pathways. Comparing the genomic profiles of 14 TC-GSC pairs, we evidenced a robust similarity among the two samples of each patient. The shared imbalanced genes are related to the development and progression of cancer and in metabolic pathways, as shown by bioinformatic analysis using DAVID. Finally, the comparison between 7 TC-PBZ pairs leads to the identification of PBZ-unique alterations that require further investigation.

Not expected

Background. Altitude training has been shown to improve endurance and ultra-en-durance performance at altitude, whereas the possible benefits from altitude/hypoxic training for competing at sea level have been, and still are, a matter for debate. Reasons for this discrepancy may result from the variety of protocols utilized in terms of alti-tude, natural or simulated, to which the athletes were exposed, and amount of the time spent at altitude. In order to conciliate previous findings and provide practical recommendations to athletes, the concept of optimal “hypoxic dose” has been defined. Methods. To perform a review of the literature concerning the effects of altitude training on athletic performance. Results. The dominant paradigm is that the improved performance at sea level is due primarily to an accelerated erythropoiesis due to the reduced oxygen available at alti-tude, leading to an increase in red cell mass. Indeed, in recent years it has become evident that other non-hematological factors (improved muscle efficiency, greater muscle buffering capacity, etc.), may contribute to improve athletic performance. Conclusions. Despite more than fifty years of research and studies, altitude training remains a controversial issue and yet, there are many unanswered questions.

No abstract

During the last decade, there has been a growing interest in understanding the fate of food during digestion in the gastrointestinal tract in order to strengthen the possible effects of food on human health. Ideally, food digestion should be studied in vivo on humans but this is not always ethically and financially possible. Therefore simple static in vitro digestion models mimicking the gastrointestinal tract have been proposed as alternatives to in vivo experiments but these models are quite basic and hardly recreate the complexity of the digestive tract. In contrast, dynamic models that allow pH regulation, flow of the food and injection in real time of digestive enzymes in the different compartments of the gastrointestinal tract are more promising to accurately mimic the digestive process. Most of the systems developed so far have been compared for their performances to in vivo data obtained on animals and/or humans. The objective of this article is to review the validation towards in vivo data of some of the dynamic digestion systems currently available in order to determine what aspects of food digestion they are able to mimic. Eight dynamic digestion systems are presented as well as their validation towards in vivo data. Advantages and limits of each simulator is discussed. This is the result of a cooperative international effort made by some of the scientists involved in Infogest, an international network on food digestion.

Our aim was to investigate the relationship between physiological variables (not previously studied) and performance in elite 1,500-m runners. We assessed eight male athletes with an average personal best time of 233.3 ± 6.9 s (110% of the world record) for the 1,500-m race. Ventilatory measurements, maximal oxygen consumption VO2max maximal vastus lateralis muscle deoxygenation (∆[deoxy(Hb+Mb)])max via near-infrared spectroscopy (NIRS), and maximal velocity (V (max)) were obtained during an incremental treadmill test. During subsequent constant-speed exercise at Vmax, we determined the time to exhaustion (Tlim), end-exercise blood lactate concentration ([La]b(max)), VO2 and ∆[deoxy(Hb+Mb)] kinetics parameters. The mean VO2max, [La]b(max) and Vmax were 70.2 ± 3.9 mL kg(-1) min(-1), 12.7 ± 2.4 mmol L(-1), 21.5 ± 0.5 km h(-1), respectively. VO2 at Vmax showed a significant negative correlation with Tlim, whereas [La]b(max) was positively correlated with Tlim. Race speed was found to significantly correlate with ∆[deoxy(Hb+Mb)](max) (79% of maximal value obtained during a transient limb ischemia), ∆[deoxy(Hb+Mb)] slow component (22.9 ± 9.3% of total amplitude) and [La]b(max) at Vmax. [La]b(max) at Vmax was also significantly correlated with ∆[deoxy(Hb+Mb)] slow component, suggesting a greater release of oxygen from the hemoglobin due to the Bohr effect. We conclude that both the maximal capacity of muscle to extract O2 from the blood and the end-exercise blood lactate accumulation are important predictors of best performance in 1,500-m runners.

Porcelli S, Marzorati M, Lanfranconi F, Vago P, Pisot R, Grassi B. Role of skeletal muscles impairment and brain oxygenation in limiting oxidative metabolism during exercise after bed rest. J Appl Physiol 109: 101-111, 2010. First published April 15, 2010; doi:10.1152/japplphysiol.00782.2009.-"Central" and "peripheral" limitations to oxidative metabolism during exercise were evaluated in 10 young males following a 35-day horizontal bed rest (BR). Incremental exercise (IE) and moderate-and heavy-intensity constant-load exercises (CLE) were carried out on a cycloergometer before and 1-2 days after BR. Pulmonary gas exchange, cardiac output (Q.; by impedance cardiography), skeletal muscle (vastus lateralis), and brain (frontal cortex) oxygenation (by near-infrared spectroscopy) were determined. After BR, "peak" (values at exhaustion during IE) workload, peak O-2 uptake ((V) over dotO(2peak)), peak stroke volume, (Q) over dot(peak), and peak skeletal muscle O-2 extraction were decreased (-18, -18, -22, -19, and -33%, respectively). The gas exchange threshold was similar to 60% of (V) over dotO(2peak) both before and after BR. At the highest workloads, brain oxygenation data suggest an increased O-2 extraction, which was unaffected by BR. (V) over dotO(2) kinetics during CLE (same percentage of peak workload before and after BR) were slower (time constant of the "fundamental" component: 31.1 +/- 2.0 s before vs. 40.0 +/- 2.2 s after BR); the amplitude of the "slow component" was unaffected by BR, thus it would be greater, after BR, at the same absolute workload. A more pronounced "overshoot" of skeletal muscle O-2 extraction during CLE was observed after BR, suggesting an impaired adjustment of skeletal muscle O-2 delivery. The role of skeletal muscles in the impairment of oxidative metabolism during submaximal and maximal exercise after BR was identified. The reduced capacity of peak cardiovascular O-2 delivery did not determine a "competition" for the available O-2 bet

Dynamic, multicompartment in vitro gastrointestinal simulators are often used to monitor gut microbial dynamics and activity. These reactors need to harbor a microbial community that is stable upon inoculation, colon region specific, and relevant to in vivo conditions. Together with the reproducibility of the colonization process, these criteria are often overlooked when the modulatory properties from different treatments are compared. We therefore investigated the microbial colonization process in two identical simulators of the human intestinal microbial ecosystem (SHIME), simultaneously inoculated with the same human fecal microbiota with a high-resolution phylogenetic microarray: the human intestinal tract chip (HITChip). Following inoculation of the in vitro colon compartments, microbial community composition reached steady state after 2 weeks, whereas 3 weeks were required to reach functional stability. This dynamic colonization process was reproducible in both SHIME units and resulted in highly diverse microbial communities which were colon region specific, with the proximal regions harboring saccharolytic microbes (e.g., Bacteroides spp. and Eubacterium spp.) and the distal regions harboring mucin-degrading microbes (e.g., Akkermansia spp.). Importantly, the shift from an in vivo to an in vitro environment resulted in an increased Bacteroidetes/Firmicutes ratio, whereas Clostridium cluster IX (propionate producers) was enriched compared to clusters IV and XIVa (butyrate producers). This was supported by proportionally higher in vitro propionate concentrations. In conclusion, high-resolution analysis of in vitro-cultured gut microbiota offers new insight on the microbial colonization process and indicates the importance of digestive parameters that may be crucial in the development of new in vitro models

Urania basin in the deep Mediterranean Sea houses a lake that is >100 m deep, devoid of oxygen, 6 times more saline than seawater, and has very high levels of methane and particularly sulfide (up to 16 mM), making it among the most sulfidic water bodies on Earth. Along the depth profile there are 2 chemoclines, a steep one with the overlying oxic seawater, and another between anoxic brines of different density, where gradients of salinity, electron donors and acceptors occur. To identify and differentiate the microbes and processes contributing to the turnover of organic matter and sulfide along the water column, these chemoclines were sampled at a high resolution. Bacterial cell numbers increased up to a hundredfold in the chemoclines as a consequence of elevated nutrient availability, with higher numbers in the upper interface where redox gradient was steeper. Bacterial and archaeal communities, analyzed by DNA fingerprinting, 16S rRNA gene libraries, activity measurements, and cultivation, were highly stratified and metabolically more active along the chemoclines compared with seawater or the uniformly hypersaline brines. Detailed analysis of 16S rRNA gene sequences revealed that in both chemoclines - and -Proteobacteria, predominantly sulfate reducers and sulfur oxidizers, respectively, were the dominant bacteria. In the deepest layers of the basin MSBL1, putatively responsible for methanogenesis, dominated among archaea. The data suggest that the complex microbial community is adapted to the basin’s extreme chemistry, and the elevated biomass is driven largely by sulfur cycling and methanogenesis

Urania basin in the deep Mediterranean Sea houses a lake that is > 100 m deep, devoid of oxygen, 6 times more saline than seawater, and has very high levels of methane and particularly sulfide (up to 16 mM), making it among the most sulfidic water bodies on Earth. Along the depth profile there are 2 chemoclines, a steep one with the overlying oxic seawater, and another between anoxic brines of different density, where gradients of salinity, electron donors and acceptors occur. To identify and differentiate the microbes and processes contributing to the turnover of organic matter and sulfide along the water column, these chemoclines were sampled at a high resolution. Bacterial cell numbers increased up to a hundredfold in the chemoclines as a consequence of elevated nutrient availability, with higher numbers in the upper interface where redox gradient was steeper. Bacterial and archaeal communities, analyzed by DNA fingerprinting, 16S rRNA gene libraries, activity measurem! ents, and cultivation, were highly stratified and metabolically more active along the chemoclines compared with seawater or the uniformly hypersaline brines. Detailed analysis of 16S rRNA gene sequences revealed that in both chemoclines delta- and epsilon-Proteobacteria, predominantly sulfate reducers and sulfur oxidizers, respectively, were the dominant bacteria. In the deepest layers of the basin MSBL1, putatively responsible for methanogenesis, dominated among archaea. The data suggest that the complex microbial community is adapted to the basin's extreme chemistry, and the elevated biomass is driven largely by sulfur cycling and methanogenesis., Urania basin in the deep Mediterranean Sea houses a lake that is > 100 m deep, devoid of oxygen, 6 times more saline than seawater, and has very high levels of methane and particularly sulfide (up to 16 mM), making it among the most sulfidic water bodies on Earth. Along the depth profile there are 2 chemoclines, a steep one with the overlying oxic seawater, and another between anoxic brines of different density, where gradients of salinity, electron donors and acceptors occur. To identify and differentiate the microbes and processes contributing to the turnover of organic matter and sulfide along the water column, these chemoclines were sampled at a high resolution. Bacterial cell numbers increased up to a hundredfold in the chemoclines as a consequence of elevated nutrient availability, with higher numbers in the upper interface where redox gradient was steeper. Bacterial and archaeal communities, analyzed by DNA fingerprinting, 16S rRNA gene libraries, activity measurem! ents, and cultivation, were highly stratified and metabolically more active along the chemoclines compared with seawater or the uniformly hypersaline brines. Detailed analysis of 16S rRNA gene sequences revealed that in both chemoclines delta- and epsilon-Proteobacteria, predominantly sulfate reducers and sulfur oxidizers, respectively, were the dominant bacteria. In the deepest layers of the basin MSBL1, putatively responsible for methanogenesis, dominated among archaea. The data suggest that the complex microbial community is adapted to the basin's extreme chemistry, and the elevated biomass is driven largely by sulfur cycling and methanogenesis.

Purpose: The aim was to identify additional noninvasive tools allowing to detect and to quantify the metabolic impairment in patients with mitochondrial myopathies (MM) or McArdle's disease (McA). Methods: Kinetics of adjustment of pulmonary oxygen uptake ((V) over dotO(2) kinetics) during transitions to constant-load moderate-intensity cycle ergometer exercise were determined on 15 MM, 8 McA, 21 patients with signs and/or symptoms of metabolic myopathy but a negative biopsy ("patient controls"; P-CTRL), and 22 healthy untrained controls (CTRL). Results: (V) over dotO(2) kinetics were slower in MM and in McA versus P-CTRL and CTRL, slower in McA versus MM, and not significantly different between P-CTRL and CTRL. The time constants (tau) of the monoexponential function describing the (V) over dotO(2) kinetics were ((X) over bar +/- SE) 59.2 +/- 8.5 s in MM, 87.6 +/- 16.4 s in McA, 36.9 +/- 3.1 s in P-CTRL, and 35.4 +/- 1.9 s in CTRL. In a subgroup of the patients (eight MM and seven McA), tau of (V) over dotO(2) kinetics were negatively correlated with two variables determined in a previous study (Grassi B, Marzorati M, Lanfranconi F, et al. Impaired oxygen extraction in metabolic myopathies: detection and quantification by near-infrared spectroscopy. Muscle Nerve. 2007;35:510-20): a) a muscle oxygenation index, obtained by near-infrared spectroscopy, estimating the peak capacity of skeletal muscle fractional O(2) extraction; and b) (V) over dotO(2) peak. Conclusions: In MM and McA patients, analysis of pulmonary (V) over dotO(2) kinetics during moderate-intensity exercise allows to identify and to quantify, noninvasively, the impairment of skeletal muscle oxidative metabolism. In these patients, the slower (V) over dotO(2) kinetics can be considered a marker of the impaired exercise tolerance. The present data could be useful for clinicians who need an objective, quantitative, and longitudinal evaluation of the impairment to be used in the follow-up of these patient

Scaphoideus titanus is the insect vector of flavescence dorée (FD), a yellow disease of grapevines. Observations on adult females and nymphs of S. titanus showed that this insect is associated with a complex microbial community. Ultrastructural analysis showed that the fat body, salivary glands and ovary of the insect harbour microorganisms showing the brush-like structure typically observed in the genus Cardinium. In particular, it has been shown that these symbiotic bacteria are present both in the follicular cells and in the eggs. In addition, cells resembling bacteriocytes, harbouring numerous Cardinium symbionts in the cytoplasm, were observed in the apical portion of the ovary in adult females. These cells are likely responsible for bacterial transmission to the ovary. Optical microscopy showed that the fat body harbours an enormous population of yeast-like symbionts (YLSs). Ultrastructural observations showed that these symbionts are enclosed within specialized cells of the fat body and are also present in the ovary, where they are found in both the follicular cells and the eggs. There is thus evidence that both Cardinium and the YLSs are transovarially transmitted to the offspring. To our knowledge, S. titanus is the sole insect known to transmit two different kinds of symbionts to the eggs, a prokaryote and an eukaryote. Gene sequence analysis and in situ hybridization led to the identification of YLSs as members of the class Sordariomycetes (=Pyrenomycetes). Finally, ultrastructural observation of the midgut content revealed the presence, in both adult females and nymphs, of a complex microbial community, which include a phytoplasma-like microorganism, likely the agent of FD.

Grape yellows caused by phytoplasmas afflict several important wine-producing areas of Europe. A grape yellows with increasing incidence in European vineyards is “bois noir” (BN), caused by ‘Candidatus Phytoplasma solani’. Its vector is the planthopper Hyalesthes obsoletus Signoret (Hemiptera Cixiidae), occasionally feeding on grapevine. An innovative strategy for reducing the diffusion of the disease could be symbiotic control, exploiting the action of symbiotic microorganisms of the insect host. To investigate the occurrence of possible microbial candidates for symbiotic control we performed a molecular characterization of the bacteria associated to H. obsoletus. Length heterogeneity PCR was applied for a preliminary population screening. Taxonomic affiliations of the bacterial species were analyzed by denaturing gradient gel electrophoresis, showing, within the microbial diversity, the intracellular reproductive parasite Wolbachia pipientis and a Bacteroidetes symbiont with 92% nt identity with ‘Candidatus Sulcia muelleri’. PCR essays specific for these bacteria showed they co-localize in several organs of H. obsoletus. Fluorescent in situ hybridization was performed to assess the distribution of these microorganisms within the insect body, showing interesting localization patterns, particularly in insect gonads and salivary glands. These results could be a starting point for a deeper investigation of functions and relationships between microbial species.

Scaphoideus titanus Ball (Hemiptera Cicadellidae) is a phloem-feeder leafhopper vector of ‘Candidatus Phytoplasma vitis’, the phytoplasma responsible of “flavescence dorée” (FD), a grape yellows afflicting several wine production areas in Europe. Natural symbionts of the leafhopper for symbiotic control of FD transmission could be used. Previous work showed the presence of a CFB bacterium, ‘Candidatus Cardinium sp.’ (named ST1-C) in several body districts of S. titanus. Our work was aimed to investigate the life cycle of ST1-C endosymbiont inside and outside of insect body. We fed S. titanus individuals on artificial media and tested insects and diets after the treatment for the presence of the ST1-C endosymbiont by qualitative and quantitative PCR, showing the release of the bacterium during feeding. We also performed fluorescent in situ hybridization on the sugar solutions after insect feeding, detecting ST1-C cells that confirmed its release into the medium. We tested by ST1-C-specific PCR DNA extracted from small areas of grape leaves exposed to S. titanus, identifying the bacterium in the majority of pierced leaves. Our experiments suggest that the ST1-C endosymbiont can undergo a horizontal transmission and have a complex life cycle possibly involving more than one host.

A recent survey on the microflora associated with the leafhopper Scaphoideus titanus Ball (Hemiptera Cicadellidae), the vector of “flavescence dorée” (FD), showed the presence of bacteria belonging to the genus Asaia. These bacteria are symbionts of insects of the genus Anopheles, malarial mosquito vectors. We focused on the association between Asaia sp. and S. titanus with the aim to evaluate a possible role of this microorganism as a symbiotic control agent. After development of artificial feeding systems for S. titanus, quantitative PCR on insects or feeds DNA was performed with specific primers. Results underlined that Asaia sp. was dominant in the insect microflora. A strain of Asaia expressing a green fluorescent protein was used to perform recolonization of the body of S. titanus, showing the colonization by the bacterium of salivary glands, guts, female and male reproductive organs of the insect. In situ hybridization with specific probes was performed on insect dissections, confirming the presence of the bacterium in spermatic bundles and in Malpighian tubules. Easy cultivability, dominance within the bacterial population in the insect body, cryogenic preservability and easiness for genetic manipulation demonstrated by the overall data make Asaia sp. an optimal candidate for carrying factors for FD control.

Here, we show that an alpha-proteobacterium of the genus Asaia is stably associated with larvae and adults of Anopheles stephensi, an important mosquito vector of Plasmodium vivax, a main malaria agent in Asia. Asaia bacteria dominate mosquito-associated microbiota, as shown by 16S rRNA gene abundance, quantitative PCR, transmission electron microscopy and in situ-hybridization of 16S rRNA genes. In adult mosquitoes, Asaia sp. is present in high population density in the female gut and in the male reproductive tract. Asaia sp. from An. stephensi has been cultured in cell-free media and then transformed with foreign DNA. A green fluorescent protein-tagged Asaia sp. strain effectively lodged in the female gut and salivary glands, sites that are crucial for Plasmodium sp. development and transmission. The larval gut and the male reproductive system were also colonized by the transformed Asaia sp. strain. As an efficient inducible colonizer of mosquitoes that transmit Plasmodium sp., Asaia sp. may be a candidate for malaria control.