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1.
Arch Microbiol ; 205(2): 73, 2023 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-36705763

RESUMO

Over the last decades, losses of bee populations have been observed worldwide. A panoply of biotic and abiotic factors, as well as the interplay among them, has been suggested to be responsible for bee declines, but definitive causes have not yet been identified. Among pollinators, the honeybee Apis mellifera is threatened by various diseases and environmental stresses, which have been shown to impact the insect gut microbiota that is known to be fundamental for host metabolism, development and immunity. Aimed at preserving the gut homeostasis, many researches are currently focusing on improving the honeybee health through the administration of probiotics e.g., by boosting the innate immune response against microbial infections. Here, we review the knowledge available on the characterization of the microbial diversity associated to honeybees and the use of probiotic symbionts as a promising approach to maintain honeybee fitness, sustaining a healthy gut microbiota and enhancing its crucial relationship with the host immune system.


Assuntos
Microbioma Gastrointestinal , Probióticos , Abelhas , Animais , Probióticos/uso terapêutico , Imunidade Inata , Gerenciamento Clínico
2.
Microorganisms ; 10(12)2022 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-36557733

RESUMO

Flavonoids are a broad class of secondary metabolites with multifaceted functionalities for plant homeostasis and are involved in facing both biotic and abiotic stresses to sustain plant growth and health. Furthermore, they were discovered as mediators of plant networking with the surrounding environment, showing a surprising ability to perform as signaling compounds for a multitrophic inter-kingdom level of communication that influences the plant host at the phytobiome scale. Flavonoids orchestrate plant-neighboring plant allelopathic interactions, recruit beneficial bacteria and mycorrhizal fungi, counteract pathogen outbreak, influence soil microbiome and affect plant physiology to improve its resilience to fluctuating environmental conditions. This review focuses on the diversified spectrum of flavonoid functions in plants under a variety of stresses in the modulation of plant morphogenesis in response to environmental clues, as well as their role as inter-kingdom signaling molecules with micro- and macroorganisms. Regarding the latter, the review addresses flavonoids as key phytochemicals in the human diet, considering their abundance in fruits and edible plants. Recent evidence highlights their role as nutraceuticals, probiotics and as promising new drugs for the treatment of several pathologies.

3.
Antibiotics (Basel) ; 11(9)2022 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-36140010

RESUMO

Antibiotic resistance spread must be considered in a holistic framework which comprises the agri-food ecosystems, where plants can be considered a bridge connecting water and soil habitats with the human microbiome. However, the study of horizontal gene transfer events within the plant microbiome is still overlooked. Here, the environmental strain Acinetobacter baylyi BD413 was used to study the acquisition of extracellular DNA (exDNA) carrying an antibiotic resistance gene (ARG) on lettuce phylloplane, performing experiments at conditions (i.e., plasmid quantities) mimicking those that can be found in a water reuse scenario. Moreover, we assessed how the presence of a surfactant, a co-formulant widely used in agriculture, affected exDNA entry in bacteria and plant tissues, besides the penetration and survival of bacteria into the leaf endosphere. Natural transformation frequency in planta was comparable to that occurring under optimal conditions (i.e., temperature, nutrient provision, and absence of microbial competitors), representing an entrance pathway of ARGs into an epiphytic bacterium able to penetrate the endosphere of a leafy vegetable. The presence of the surfactant determined a higher presence of culturable transformant cells in the leaf tissues but did not significantly increase exDNA entry in A. baylyi BD413 cells and lettuce leaves. More research on HGT (Horizontal Gene Transfer) mechanisms in planta should be performed to obtain experimental data on produce safety in terms of antibiotic resistance.

4.
AMB Express ; 12(1): 98, 2022 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-35895126

RESUMO

Macroalgal surface constitutes a peculiar ecological niche and an advantageous substratum for microorganisms able to degrade the wide diversity of algal glycans. The degrading enzymatic activities of macroalgal epiphytes are of paramount interest for the industrial by-product sector and biomass resource applications. We characterized the polysaccharide hydrolytic profile of bacterial isolates obtained from three macroalgal species: the red macroalgae Asparagopsis taxiformis and Sphaerococcus coronopifolius (Rhodophyceae) and the brown Halopteris scoparia (Phaeophyceae), sampled in South Portugal. Bacterial enrichment cultures supplemented with chlorinated aliphatic compounds, typically released by marine algae, were established using as inoculum the decaying biomass of the three macroalgae, obtaining a collection of 634 bacterial strains. Although collected from the same site and exposed to the same seawater seeding microbiota, macroalgal cultivable bacterial communities in terms of functional and phylogenetic diversity showed host specificity. Isolates were tested for the hydrolysis of starch, pectin, alginate and agar, exhibiting a different hydrolytic potential according to their host: A. taxiformis showed the highest percentage of active isolates (91%), followed by S. coronopifolius (54%) and H. scoparia (46%). Only 30% of the isolates were able to degrade starch, while the other polymers were degraded by 55-58% of the isolates. Interestingly, several isolates showed promiscuous capacities to hydrolyze more than one polysaccharide. The isolate functional fingerprint was statistically correlated to bacterial phylogeny, host species and enrichment medium. In conclusion, this work depicts macroalgae as holobionts with an associated microbiota of interest for blue biotechnologies, suggesting isolation strategies and bacterial targets for polysaccharidases' discovery.

5.
Appl Environ Microbiol ; 88(9): e0252221, 2022 05 10.
Artigo em Inglês | MEDLINE | ID: mdl-35416683

RESUMO

This study shows that Escherichia coli can be temporarily enriched in zooplankton under natural conditions and that these bacteria can belong to different phylogroups and sequence types (STs), including environmental, clinical, and animal isolates. We isolated 10 E. coli strains and sequenced the genomes of two of them. Phylogenetically, the two isolates were closer to strains isolated from poultry meat than to freshwater E. coli, albeit their genomes were smaller than those of the poultry isolates. After isolation and fluorescent protein tagging of strains ED1 and ED157, we show that Daphnia sp. can take up these strains and release them alive again, thus becoming a temporary host for E. coli. In a chemostat experiment, we show that this association does not prolong bacterial long-term survival, but at low abundances it also does not significantly reduce bacterial numbers. We demonstrate that E. coli does not belong to the core microbiota of Daphnia, suffers from competition by the natural Daphnia microbiota, but can profit from its carapax to survive in water. All in all, this study suggests that the association of E. coli with Daphnia is only temporary, but the cells are viable therein, and this might allow encounters with other bacteria for genetic exchange and potential genomic adaptation to the freshwater environment. IMPORTANCE The contamination of freshwater with feces-derived bacteria is a major concern regarding drinking water acquisition and recreational activities. Ecological interactions promoting their persistence are still very scarcely studied. This study, which analyses the survival of E. coli in the presence of zooplankton, is thus of ecological and water safety relevance.


Assuntos
Água Potável , Escherichia coli , Animais , Bactérias , Daphnia/microbiologia , Escherichia coli/genética , Fezes/microbiologia , Água Doce/microbiologia , Zooplâncton/microbiologia
6.
Microbiol Spectr ; 10(1): e0158021, 2022 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-34985334

RESUMO

Stress-associated dysbiosis of microbiome can have several configurations that, under an energy landscape conceptual framework, can change from one configuration to another due to different alternating selective forces. It has been proposed-according to the Anna Karenina Principle-that in stressed individuals the microbiome are more dispersed (i.e., with a higher within-beta diversity), evidencing the grade of dispersion as indicator of microbiome dysbiosis. We hypothesize that although dysbiosis leads to different microbial communities in terms of beta diversity, these are not necessarily differently dispersed (within-beta diversity), but they form disrupted networks that make them less resilient to stress. To test our hypothesis, we select nutrient restriction (NR) stress that impairs host fitness but does not introduce overt microbiome selectors, such as toxic compounds and pathogens. We fed the polyphagous black soldier fly, Hermetia illucens, with two NR diets and a control full-nutrient (FN) diet. NR diets were dysbiotic because they strongly affected insect growth and development, inducing significant microscale changes in physiochemical conditions of the gut compartments. NR diets established new configurations of the gut microbiome compared to FN-fed guts but with similar dispersion. However, these new configurations driven by the deterministic changes induced by NR diets were reflected in rarefied, less structured, and less connected bacterial interactomes. These results suggested that while the dispersion cannot be considered a consistent indicator of the unhealthy state of dysbiotic microbiomes, the capacity of the community members to maintain network connections and stability can be an indicator of the microbial dysbiotic conditions and their incapacity to sustain the holobiont resilience and host homeostasis. IMPORTANCE Changes in diet play a role in reshaping the gut microbiome in animals, inducing dysbiotic configurations of the associated microbiome. Although studies have reported on the effects of specific nutrient contents on the diet, studies regarding the conditions altering the microbiome configurations and networking in response to diet changes are limited. Our results showed that nutrient poor diets determine dysbiotic states of the host with reduction of insect weight and size, and increase of the times for developmental stage. Moreover, the poor nutrient diets lead to changes in the compositional diversity and network interaction properties of the gut microbial communities. Our study adds a new component to the understanding of the ecological processes associated with dysbiosis, by disentangling consequences of diets on microbiome dysbiosis that is manifested with the disruption of microbiome networking properties rather than changes in microbiome dispersion and beta diversity.


Assuntos
Ração Animal/análise , Bactérias/isolamento & purificação , Microbioma Gastrointestinal , Nutrientes/metabolismo , Simuliidae/microbiologia , Animais , Bactérias/classificação , Bactérias/genética , Biodiversidade , Nutrientes/análise , Simuliidae/crescimento & desenvolvimento , Simuliidae/metabolismo
7.
Materials (Basel) ; 14(13)2021 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-34279292

RESUMO

The increasing demand for reconstructions of concrete structures and the wide availability on the market of surface protective products and systems could lead to misunderstandings in the decision of the most effective solution. Surface protectors have become increasingly widespread in recent years in concrete restoration interventions thanks to their properties: they are able to protect the substrate from aggressive agents and consequently extend the useful life of the structures. The aim of this article is first of all to present the surface protective treatments available on the market, outlining their strengths and weaknesses. Subsequently, a characterization of seven different commercial coatings for reinforced-concrete structures is provided, taking into account chemical nature, fields of use and effectiveness, both in terms of physic and elastic performance and resistance to aggressive agents that undermine the durability of the treated concrete elements.

8.
NPJ Biofilms Microbiomes ; 7(1): 42, 2021 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-33963194

RESUMO

The core gut microbiome of adult honeybee comprises a set of recurring bacterial phylotypes, accompanied by lineage-specific, variable, and less abundant environmental bacterial phylotypes. Several mutual interactions and functional services to the host, including the support provided for growth, hormonal signaling, and behavior, are attributed to the core and lineage-specific taxa. By contrast, the diversity and distribution of the minor environmental phylotypes and fungal members in the gut remain overlooked. In the present study, we hypothesized that the microbial components of forager honeybees (i.e., core bacteria, minor environmental phylotypes, and fungal members) are compartmentalized along the gut portions. The diversity and distribution of such three microbial components were investigated in the context of the physico-chemical conditions of different gut compartments. We observed that changes in the distribution and abundance of microbial components in the gut are consistently compartment-specific for all the three microbial components, indicating that the ecological and physiological interactions among the host and microbiome vary with changing physico-chemical and metabolic conditions of the gut.


Assuntos
Bactérias , Abelhas , Biodiversidade , Fungos , Microbioma Gastrointestinal , Animais , Bactérias/classificação , Bactérias/ultraestrutura , Fungos/classificação , Fungos/ultraestrutura , Metagenoma , Metagenômica/métodos
9.
Front Microbiol ; 11: 574301, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33013812

RESUMO

The diffusion of antibiotic resistance determinants in different environments, e.g., soil and water, has become a public concern for global health and food safety and many efforts are currently devoted to clarify this complex ecological and evolutionary issue. Horizontal gene transfer (HGT) has an important role in the spread of antibiotic resistance genes (ARGs). However, among the different HGT mechanisms, the capacity of environmental bacteria to acquire naked exogenous DNA by natural competence is still poorly investigated. This study aimed to characterize the ability of the environmental Escherichia coli strain ED1, isolated from the crustacean Daphnia sp., to acquire exogenous DNA by natural competence. Transformation experiments were carried out varying different parameters, i.e., cell growth phase, amount of exogenous DNA and exposition to artificial lake water (ALW) and treated wastewater to mimic environmental-like conditions that may be encountered in the agri-food system. Results were compared with those showed by the laboratory E. coli strain DH5α. Our experimental data, supported by genomic sequencing, showed that, when exposed to pure water, ED1 strain was able to acquire exogenous DNA with frequencies (10-8-10-9) statistically higher than the ones observed for DH5α strain (10-10). Interestingly, higher values were retrieved for ED1 than DH5α strains exposed to ALW (10-7 vs. 10-9, respectively) or treated wastewater (10-8 vs. 10-10, respectively). We tested, therefore, ED1 strain ability to colonize the rhizosphere of lettuce, a model plant representative of raw-consumed vegetables of high economic importance in the ready-to-eat food industry. Results showed that ED1 strain was able to efficiently colonize lettuce rhizosphere, revealing a stable colonization for 14 days-long period. In conclusion, ED1 strain ability to acquire exogenous DNA in environmental-like conditions by natural competence, combined with its ability to efficiently and stably colonize plant rhizosphere, poses the attention to food and human safety showing a possible route of diffusion of antibiotic resistance in the agri-food system, sustaining the "One Health" warnings related to the antibiotic spread.

10.
Front Microbiol ; 11: 1965, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32903451

RESUMO

Larvae of the black soldier fly (BSF) Hermetia illucens (L.) convert organic waste into high valuable insect biomass that can be used as alternative protein source for animal nutrition or as feedstock for biodiesel production. Since insect biology and physiology are influenced by the gut microbiome, knowledge about the functional role of BSF-associated microorganisms could be exploited to enhance the insect performance and growth. Although an increasing number of culture-independent studies are unveiling the microbiota structure and composition of the BSF gut microbiota, a knowledge gap remains on the experimental validation of the contribution of the microorganisms to the insect growth and development. We aimed at assessing if BSF gut-associated bacteria potentially involved in the breakdown of diet components are able to improve host nutrition. A total of 193 bacterial strains were obtained from guts of BSF larvae reared on a nutritious diet using selective and enrichment media. Most of the bacterial isolates are typically found in the insect gut, with major representatives belonging to the Gammaproteobacteria and Bacilli classes. The hydrolytic profile of the bacterial collection was assessed on compounds typically present in the diet. Finally, we tested the hypothesis that the addition to a nutritionally poor diet of the two isolates Bacillus licheniformis HI169 and Stenotrophomonas maltophilia HI121, selected for their complementary metabolic activities, could enhance BSF growth. B. licheniformis HI169 positively influenced the larval final weight and growth rate when compared to the control. Conversely, the addition of S. maltophilia HI121 to the nutritionally poor diet did not result in a growth enhancement in terms of larval weight and pupal weight and length in comparison to the control, whereas the combination of the two strains positively affected the larval final weight and the pupal weight and length. In conclusion, we isolated BSF-associated bacterial strains with potential positive properties for the host nutrition and we showed that selected isolates may enhance BSF growth, suggesting the importance to evaluate the effect of the bacterial administration on the insect performance.

11.
Sci Rep ; 10(1): 13150, 2020 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-32753591

RESUMO

Virtual screening techniques and in vitro binding/inhibitory assays were used to search within a set of more than 8,000 naturally occurring small ligands for candidate inhibitors of 8-hydroxy-5-deazaflavin:NADPH oxidoreductase (FNO) from Methanobrevibacter smithii, the enzyme that catalyses the bidirectional electron transfer between NADP+ and F420H2 during the intestinal production of CH4 from CO2. In silico screening using molecular docking classified the ligand-enzyme complexes in the range between - 4.9 and - 10.5 kcal/mol. Molecular flexibility, the number of H-bond acceptors and donors, the extent of hydrophobic interactions, and the exposure to the solvent were the major discriminants in determining the affinity of the ligands for FNO. In vitro studies on a group of these ligands selected from the most populated/representative clusters provided quantitative kinetic, equilibrium, and structural information on ligands' behaviour, in optimal agreement with the predictive computational results.


Assuntos
Proteínas de Bactérias , Inibidores Enzimáticos/química , Methanobrevibacter/enzimologia , NADH NADPH Oxirredutases , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/química , Domínio Catalítico , NADH NADPH Oxirredutases/antagonistas & inibidores , NADH NADPH Oxirredutases/química
12.
Materials (Basel) ; 13(8)2020 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-32344730

RESUMO

One of the most important parameters concerning durability is undoubtedly represented by cement matrix resistance to chloride diffusion in environments where reinforced concrete structures are exposed to the corrosion risk induced by marine environment or de-icing salts. This paper deals with protection from chloride ingress by a silane-based surface-applied corrosion inhibitor. Results indicated that the corrosion inhibitor (CI) allows to reduce the penetration of chloride significantly compared to untreated specimens, independently of w/c, cement type, and dosage. Reduction of chloride diffusion coefficient (Dnssn) measured by an accelerated test in treated concrete was in the range 30-60%. Natural chloride diffusion test values indicate a sharp decrease in apparent diffusion coefficient (Dapp) equal to about 75% when concrete is protected by CI. Mechanism of action of CI in slowing down the chloride penetration inside the cement matrix is basically due to the water repellent effect as confirmed by data of concrete bulk electrical resistivity.

13.
Commun Biol ; 3(1): 105, 2020 03 06.
Artigo em Inglês | MEDLINE | ID: mdl-32144396

RESUMO

Wolbachia can reduce the capability of mosquitoes to transmit infectious diseases to humans and is currently exploited in campaigns for the control of arboviruses, like dengue and Zika. Under the assumption that Wolbachia-mediated activation of insect immunity plays a role in the reduction of mosquito vectorial capacity, we focused our attention on the Wolbachia surface protein (WSP), a potential inductor of innate immunity. We hypothesized that the heterologous expression of this protein in gut- and tissue-associated symbionts may reduce parasite transmission. We thus engineered the mosquito bacterial symbiont Asaia to express WSP (AsaiaWSP). AsaiaWSP induced activation of the host immune response in Aedes aegypti and Anopheles stephensi mosquitoes, and inhibited the development of the heartworm parasite Dirofilaria immitis in Ae. aegypti. These results consolidate previous evidence on the immune-stimulating property of WSP and make AsaiaWSP worth of further investigations as a potential tool for the control of mosquito-borne diseases.


Assuntos
Acetobacteraceae/metabolismo , Aedes/microbiologia , Anopheles/microbiologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Dirofilaria immitis/microbiologia , Proteínas de Membrana/metabolismo , Wolbachia/metabolismo , Acetobacteraceae/genética , Aedes/imunologia , Animais , Anopheles/parasitologia , Proteínas da Membrana Bacteriana Externa/genética , Dirofilaria immitis/crescimento & desenvolvimento , Interações Hospedeiro-Parasita , Proteínas de Membrana/genética , Fagocitose , Simbiose , Wolbachia/genética
14.
Chemosphere ; 241: 124843, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31605997

RESUMO

Climate changes push for water reuse as a priority to counteract water scarcity and minimize water footprint especially in agriculture, one of the highest water consuming human activities. Phytodepuration is indicated as a promising technology for water reclamation, also in the light of its economic and ecological sustainability, and the use of specific bacterial inocula for microbial assisted phytodepuration has been proposed as a further advance for its implementation. Here we provided an overview on the selection and use of plant growth promoting bacteria in Constructed Wetland (CW) systems, showing their advantages in terms of plant growth support and pollutant degradation abilities. Moreover, CWs are also proposed for the removal of emerging organic pollutants like antibiotics from urban wastewaters. We focused on this issue, still debated in the literature, revealing the necessity to deepen the knowledge on the antibiotic resistance spread into the environment in relation to treated wastewater release and reuse. In addition, given the presence in the plant system of microhabitats (e.g. rhizosphere) that are hot spot for Horizontal Gene Transfer, we highlighted the importance of gene exchange to understand if these events can promote the diffusion of antibiotic resistance genes and antibiotic resistant bacteria, possibly entering in the food production chain when treated wastewater is used for irrigation. Ideally, this new knowledge will lead to improve the design of phytodepuration systems to maximize the quality and safety of the treated effluents in compliance with the 'One Health' concept.


Assuntos
Bactérias/metabolismo , Biodegradação Ambiental , Purificação da Água/métodos , Áreas Alagadas , Bactérias/genética , Resistência Microbiana a Medicamentos/genética , Humanos , Eliminação de Resíduos Líquidos/métodos , Águas Residuárias/análise , Águas Residuárias/química
15.
Microorganisms ; 7(10)2019 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-31554215

RESUMO

The microbiome associated with plants used in phytodepuration systems can boost plant growth and services, especially in ecosystems dealing with recalcitrant compounds, hardly removed via traditional wastewater (WW) treatments, such as azo-dyes used in textile industry. In this context, we aimed to study the cultivable microbiome selected by Phragmites australis plants in a Constructed Wetland (CW) in Morocco, in order to obtain candidate inoculants for the phytodepuration of azo-dye contaminated WW. A collection of 152 rhizospheric and endophytic bacteria was established. The strains were phylogenetically identified and characterized for traits of interest in the phytodepuration context. All strains showed Plant Growth Promotion potential in vitro and 67% of them significantly improved the growth of a model plant in vivo compared to the non bacterized control plants. Moreover, most of the isolates were able to grow in presence of several model micropollutants typically found in WW, indicating their potential use in phytodepuration of a wide spectrum of effluents. The six most promising strains of the collection were tested in CW microcosms alone or as consortium: the consortium and two single inocula demonstrated to significantly increase the removal of the model azo-dye Reactive Black 5 compared to the non bacterized controls.

16.
Environ Microbiol ; 21(11): 4343-4359, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31502415

RESUMO

Popillia japonica Newman (Coleoptera: Scarabaeidae) is a highly polyphagous invasive beetle originating from Japan. This insect is highly resilient and able to rapidly adapt to new vegetation. Insect-associated microorganisms can play important roles in insect physiology, helping their hosts to adapt to changing conditions and potentially contributing to an insect's invasive potential. Such symbiotic bacteria can be part of a core microbiota that is stably transmitted throughout the host's life cycle or selectively recruited from the environment at each developmental stage. The aim of this study was to investigate the origin, stability and turnover of the bacterial communities associated with an invasive population of P. japonica from Italy. Our results demonstrate that soil microbes represent an important source of gut bacteria for P. japonica larvae, but as the insect develops, its gut microbiota richness and diversity decreased substantially, paralleled by changes in community composition. Notably, only 16.75% of the soil bacteria present in larvae are maintained until the adult stage. We further identified the micro-environments of different gut sections as an important factor shaping microbiota composition in this species, likely due to differences in pH, oxygen availability and redox potential. In addition, P. japonica also harboured a stable bacterial community across all developmental stages, consisting of taxa well known for the degradation of plant material, namely the families Ruminococcacae, Christensenellaceae and Lachnospiraceae. Interestingly, the family Christensenallaceae had so far been observed exclusively in humans. However, the Christensenellaceae operational taxonomic units found in P. japonica belong to different taxonomic clades within this family.


Assuntos
Besouros/crescimento & desenvolvimento , Besouros/microbiologia , Microbioma Gastrointestinal , Estágios do Ciclo de Vida , Animais , Bactérias/classificação , Feminino , Masculino
17.
Front Physiol ; 10: 795, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31281266

RESUMO

Insect immunity is a crucial process in interactions between host and microorganisms and the presence of pathogenic, commensal, or beneficial bacteria may result in different immune responses. In Hemiptera vectors of phytoplasmas, infected insects are amenable to carrying high loads of phytopathogens, besides hosting other bacterial affiliates, which have evolved different strategies to be retained; adaptation to host response and immunomodulation are key aspects of insect-symbiont interactions. Most of the analyses published to date has investigated insect immune response to pathogens, whereas few studies have focused on the role of host immunity in microbiota homeostasis and vectorial capacity. Here the expression of immune genes in the leafhopper vector of phytoplasmas Euscelidius variegatus was investigated following exposure to Asaia symbiotic bacteria, previously demonstrated to affect phytoplasma acquisition by leafhoppers. The expression of four genes related to major components of immunity was measured, i.e., defensin, phenoloxidase, kazal type 1 serine protease inhibitor and Raf, a component of the Ras/Raf pathway. The response was separately tested in whole insects, midguts and cultured hemocytes. Healthy individuals were assessed along with specimens undergoing early- and late-stage phytoplasma infection. In addition, the adhesion grade of Asaia strains was examined to assess whether symbionts could establish a physical barrier against phytoplasma colonization. Our results revealed a specific activation of Raf in midguts after double infection by Asaia and flavescence dorée phytoplasma. Increased expression was observed already in early stages of phytoplasma colonization. Gut-specific localization and timing of Raf activation are consistent with the role played by Asaia in limiting phytoplasma acquisition by E. variegatus, supporting the involvement of this gene in the anti-pathogen activity. However, limited attachment capability was found for Asaia under in vitro experimental conditions, suggesting a minor contribution of physical phytoplasma exclusion from the vector gut wall. By providing evidence of immune modulation played by Asaia, these results contribute to elucidating the molecular mechanisms regulating interference with phytoplasma infection in E. variegatus. The involvement of Raf suggests that in the presence of reduced immunity (reported in Hemipterans), immune genes can be differently regulated and recruited to play additional functions, generally played by genes lost by hemipterans.

18.
Front Microbiol ; 10: 1304, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31244812

RESUMO

The adaptation of sporeformers to extreme environmental conditions is frequently questioned due to their capacity to produce highly resistant endospores that are considered as resting contaminants, not representing populations adapted to the system. In this work, in order to gain a better understanding of bacterial adaptation to extreme habitats, we investigated the phenotypic and genomic characteristics of the halophile Virgibacillus sp. 21D isolated from the seawater-brine interface (SBI) of the MgCl2-saturated deep hypersaline anoxic basin Discovery located in the Eastern Mediterranean Sea. Vegetative cells of strain 21D showed the ability to grow in the presence of high concentrations of MgCl2, such as 14.28% corresponding to 1.5 M. Biolog phenotype MicroArray (PM) was adopted to investigate the strain phenotype, with reference to carbon energy utilization and osmotic tolerance. The strain was able to metabolize only 8.4% of 190 carbon sources provided in the PM1 and PM2 plates, mainly carbohydrates, in accordance with the low availability of nutrients in its habitat of origin. By using in silico DNA-DNA hybridization the analysis of strain 21D genome, assembled in one circular contig, revealed that the strain belongs to the species Virgibacillus dokdonensis. The genome presented compatible solute-based osmoadaptation traits, including genes encoding for osmotically activated glycine-betaine/carnitine/choline ABC transporters, as well as ectoine synthase enzymes. Osmoadaptation of the strain was then confirmed with phenotypic assays by using the osmolyte PM9 Biolog plate and growth experiments. Furthermore, the neutral isoelectric point of the reconstructed proteome suggested that the strain osmoadaptation was mainly mediated by compatible solutes. The presence of genes involved in iron acquisition and metabolism indicated that osmoadaptation was tailored to the iron-depleted saline waters of the Discovery SBI. Overall, both phenomics and genomics highlighted the potential capability of V. dokdonensis 21D vegetative cells to adapt to the environmental conditions in Discovery SBI.

19.
Microbiol Res ; 223-225: 33-43, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31178049

RESUMO

Mangroves, dominating tropical intertidal zones and estuaries, are among the most salt tolerant plants, and propagate through reproductive units called propagules. Similarly to plant seeds, propagules may harbor beneficial bacteria. Our hypothesis was that mangroves, being able to grow into seawater, should harbor bacteria able to interact with the host and to exert positive effects under salt stress, which could be exploited to improve crop production. Therefore, we isolated bacterial endophytes from mangrove propagules with the aim to test whether these bacteria have a beneficial potential on their natural host and on different crops such as barley and rice, cultivated under salt stress. The 172 bacterial isolates obtained were screened for plant growth promotion (PGP) activities in vitro, and the 12 most promising isolates were tested on barley under non-axenic conditions and salt stress. Gordonia terrae KMP456-M40 was the best performing isolate, increasing ear weight by 65%. Based on the in vivo PGP activity and the root colonization ability, investigated by fluorescence in situ hybridization and confocal microscopy, three strains were additionally tested on mangrove propagule germination and on rice growth. The most effective strain was again G. terrae KMP456-M40, which enhanced the root length of mangrove seedlings and the biomass of salt-stressed rice under axenic conditions up to 65% and 62%, respectively. We demonstrated that propagules, the reproductive units of mangroves, host beneficial bacteria that enhance the potential of mangrove seedlings establishment and confer salt tolerance to cereal crops.


Assuntos
Produtos Agrícolas , Grão Comestível/crescimento & desenvolvimento , Endófitos/fisiologia , Desenvolvimento Vegetal , Estresse Salino , Bactérias/classificação , Bactérias/isolamento & purificação , Biomassa , Endófitos/classificação , Endófitos/crescimento & desenvolvimento , Endófitos/isolamento & purificação , Genótipo , Germinação , Hordeum/crescimento & desenvolvimento , Hordeum/microbiologia , Hibridização in Situ Fluorescente , Oryza/microbiologia , Pressão Osmótica , Raízes de Plantas/microbiologia , Salinidade , Tolerância ao Sal , Plantas Tolerantes a Sal/microbiologia , Arábia Saudita , Água do Mar , Plântula/crescimento & desenvolvimento , Plântula/microbiologia , Sementes/crescimento & desenvolvimento , Sementes/microbiologia , Microbiologia do Solo , Áreas Alagadas
20.
J Basic Microbiol ; 58(10): 827-835, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30019339

RESUMO

Androctonus australis is one of the most ubiquitous and common scorpion species in desert and arid lands from North Africa to India and it has an important ecological role and social impact. The bacterial community associated to this arachnid is unknown and we aimed to dissect its species composition in the gut, gonads, and venom gland. A 16S rRNA gene culture-independent diversity analysis revealed, among six other taxonomic groups (Firmicutes, Betaproteobacteria, Gammaproteobacteria, Flavobacteria, Actinobacteria, and Cyanobacteria), a dominance of Mollicutes phylotypes recorded both in the digestive tract and the gonads. These related Mollicutes include two Spiroplasma phylotypes (12.5% of DGGE bands and 15% of clones), and a new Mycoplasma cluster (80% of clones) showing 16S rRNA sequence identities of 95 and 93% with Mollicutes detected in the Mexican scorpions Centruroides limpidus and Vaejovis smithi, respectively. Such scorpion-associated Mollicutes form a new lineage that share a distant ancestor with Mycoplasma hominis. The observed host specificity with the apparent phylogenetic divergence suggests a relatively long co-evolution of these symbionts with the scorpion hosts. From the ecological point of view, such association may play a beneficial role for the host fitness, especially during dormancy or molt periods.


Assuntos
Variação Genética , Filogenia , Escorpiões/microbiologia , Simbiose , Tenericutes/classificação , Tenericutes/fisiologia , Animais , Bactérias/classificação , Bactérias/genética , Fenômenos Fisiológicos Bacterianos , DNA Bacteriano/genética , Especificidade de Hospedeiro , Índia , México , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Tenericutes/genética
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