Bacterial genome sequences of uncharacterized Chitinophaga species isolated from the International Space Station.

We report four Chitinophaga sp. strains isolated from wastewater collected onboard the International Space Station. Here, we present three finished and one draft genome. Taxonomic ranks established by genome-based analysis indicate that these Chitinophaga sp. strains represent candidates for a new species.

Genome Sequences of Bacteria Isolated from the International Space Station Water Systems.

We report draft genomes of five bacteria recovered from the U.S. and Russian water systems onboard the International Space Station. The five genera include Ralstonia, Burkholderia, Cupriavidus, Methylobacterium, and Pseudomonas. These sequences will help further the understanding of water reclamation and environmental control and life support systems in space.

Diversity of fungal microbiome obtained from plant rhizoplanes.

Microbial communities, and their ecological importance, have been investigated in several habitats. However, so far, most studies could not describe the closest microbial interactions and their functionalities. This study investigates the co-occurring interactions between fungi and bacteria in plant rhizoplanes and their potential functions. The partnerships were obtained using fungal-highway columns with four plant-based media. The fungi and associated microbiomes isolated from the columns were identified by sequencing the ITS (fungi) and 16S rRNA genes (bacteria). Statistical analyses including Exploratory Graph and Network Analysis were used to visualize the presence of underlying clusters in the microbial communities and evaluate the metabolic functions associated with the fungal microbiome (PICRUSt2). Our findings characterize the presence of both unique and complex bacterial communities associated with different fungi. The results showed that Bacillus was associated as exo-bacteria in 80 % of the fungi but occurred as putative endo-bacteria in 15 %. A shared core of putative endo-bacterial genera, potentially involved in the nitrogen cycle was found in 80 % of the isolated fungi. The comparison of potential metabolic functions of the putative endo- and exo-communities highlighted the potential essential factors to establish an endosymbiotic relationship, such as the loss of pathways associated with metabolites obtained from the host while maintaining pathways responsible for bacterial survival within the hypha.

Widespread bacterial diversity within the bacteriome of fungi.

Knowledge of associations between fungal hosts and their bacterial associates has steadily grown in recent years as the number and diversity of examinations have increased, but current knowledge is predominantly limited to a small number of fungal taxa and bacterial partners. Here, we screened for potential bacterial associates in over 700 phylogenetically diverse fungal isolates, representing 366 genera, or a tenfold increase compared with previously examined fungal genera, including isolates from several previously unexplored phyla. Both a 16 S rDNA-based exploration of fungal isolates from four distinct culture collections spanning North America, South America and Europe, and a bioinformatic screen for bacterial-specific sequences within fungal genome sequencing projects, revealed that a surprisingly diverse array of bacterial associates are frequently found in otherwise axenic fungal cultures. We demonstrate that bacterial associations with diverse fungal hosts appear to be the rule, rather than the exception, and deserve increased consideration in microbiome studies and in examinations of microbial interactions.

Democratization of fungal highway columns as a tool to investigate bacteria associated with soil fungi.

Bacteria-fungi interactions (BFIs) are essential in ecosystem functioning. These interactions are modulated not only by local nutritional conditions but also by the physicochemical constraints and 3D structure of the environmental niche. In soils, the unsaturated and complex nature of the substrate restricts the dispersal and activity of bacteria. Under unsaturated conditions, some bacteria engage with filamentous fungi in an interaction (fungal highways) in which they use fungal hyphae to disperse. Based on a previous experimental device to enrich pairs of organisms engaging in this interaction in soils, we present here the design and validation of a modified version of this sampling system constructed using additive printing. The 3D printed devices were tested using a novel application in which a target fungus, the common coprophilous fungus Coprinopsis cinerea, was used as bait to recruit and identify bacterial partners using its mycelium for dispersal. Bacteria of the genera Pseudomonas, Sphingobacterium and Stenotrophomonas were highly enriched in association with C. cinerea. Developing and producing these new easy-to-use tools to investigate how bacteria overcome dispersal limitations in cooperation with fungi is important to unravel the mechanisms by which BFIs affect processes at an ecosystem scale in soils and other unsaturated environments.

Real-Time Culture-Independent Microbial Profiling Onboard the International Space Station Using Nanopore Sequencing.

For the past two decades, microbial monitoring of the International Space Station (ISS) has relied on culture-dependent methods that require return to Earth for analysis. This has a number of limitations, with the most significant being bias towards the detection of culturable organisms and the inherent delay between sample collection and ground-based analysis. In recent years, portable and easy-to-use molecular-based tools, such as Oxford Nanopore Technologies' MinION™ sequencer and miniPCR bio's miniPCR™ thermal cycler, have been validated onboard the ISS. Here, we report on the development, validation, and implementation of a swab-to-sequencer method that provides a culture-independent solution to real-time microbial profiling onboard the ISS. Method development focused on analysis of swabs collected in a low-biomass environment with limited facility resources and stringent controls on allowed processes and reagents. ISS-optimized procedures included enzymatic DNA extraction from a swab tip, bead-based purifications, altered buffers, and the use of miniPCR and the MinION. Validation was conducted through extensive ground-based assessments comparing current standard culture-dependent and newly developed culture-independent methods. Similar microbial distributions were observed between the two methods; however, as expected, the culture-independent data revealed microbial profiles with greater diversity. Protocol optimization and verification was established during NASA Extreme Environment Mission Operations (NEEMO) analog missions 21 and 22, respectively. Unique microbial profiles obtained from analog testing validated the swab-to-sequencer method in an extreme environment. Finally, four independent swab-to-sequencer experiments were conducted onboard the ISS by two crewmembers. Microorganisms identified from ISS swabs were consistent with historical culture-based data, and primarily consisted of commonly observed human-associated microbes. This simplified method has been streamlined for high ease-of-use for a non-trained crew to complete in an extreme environment, thereby enabling environmental and human health diagnostics in real-time as future missions take us beyond low-Earth orbit.

A morphological, enzymatic and metabolic approach to elucidate apoptotic-like cell death in fungi exposed to h- and α-molybdenum trioxide nanoparticles.

The present study compares for the first time the effects of h-MoO3 and α-MoO3 against two fungal strains: Aspergillus niger and Aspergillus flavus. The h-MoO3 nanoparticles were more toxic to both fungi than α-MoO3. The toxic effects of h-MoO3 were more pronounced toward A. flavus, which presented a growth inhibition of 67.4% at 200 mg L-1. The presence of the nanoparticles affected drastically the hyphae morphology by triggering nuclear condensation and compromising the hyphae membrane. Further analysis of the volatile organic compounds (VOCs) produced by both fungi in the presence of the nanomaterials indicated important metabolic changes related to programmed cell death. These nanomaterials induced the production of specific antifungal VOCs, such as ß-Elemene and t-Cadinol, by the fungi. The production of essential enzymes involved in fungal metabolism, such as acid phosphatase, naphthol-As-BI-phosphohydrolase, ß-galactosidase, ß-glucosidase and N-acetyl-ß-glucosaminidase, reduced significantly in the presence of the nanomaterials. The changes in enzymatic production and VOCs corroborate the fact that these nanoparticles, especially h-MoO3, exert changes in the fungal metabolism, triggering apoptotic-like cell death responses in these fungi.

Recent advances in graphene-based biosensor technology with applications in life sciences.

Graphene's unique physical structure, as well as its chemical and electrical properties, make it ideal for use in sensor technologies. In the past years, novel sensing platforms have been proposed with pristine and modified graphene with nanoparticles and polymers. Several of these platforms were used to immobilize biomolecules, such as antibodies, DNA, and enzymes to create highly sensitive and selective biosensors. Strategies to attach these biomolecules onto the surface of graphene have been employed based on its chemical composition. These methods include covalent bonding, such as the coupling of the biomolecules via the 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide reactions, and physisorption. In the literature, several detection methods are employed; however, the most common is electrochemical. The main reason for researchers to use this detection approach is because this method is simple, rapid and presents good sensitivity. These biosensors can be particularly useful in life sciences and medicine since in clinical practice, biosensors with high sensitivity and specificity can significantly enhance patient care, early diagnosis of diseases and pathogen detection. In this review, we will present the research conducted with antibodies, DNA molecules and, enzymes to develop biosensors that use graphene and its derivatives as scaffolds to produce effective biosensors able to detect and identify a variety of diseases, pathogens, and biomolecules linked to diseases.

Influence of environmental factors on tenuazonic acid production by Epicoccum sorghinum: An integrative approach of field and laboratory conditions.

Sorghum is the fifth most cultivated and consumed grain in the world. However, this grain is frequently contaminated with toxins from fungi. The present study evaluated the effects of environmental factors on tenuazonic acid (TeA) production by Epicoccum sorghinum in the field and in controlled laboratory conditions. In this study, 50 sorghum grain samples were collected from summer and autumn growing seasons and analyzed for TeA contamination using LC-MS/MS. To further understand the ecophysiology of this fungus, an isolated strain of E. sorghinum from the field was investigated for its development and TeA production under controlled environmental conditions in the laboratory. In the ecophysiological investigation, the effects of water activity (0.90, 0.95, 0.99) and temperature (18, 22, 26 and 30 °C) were evaluated on the radial growth, enzymatic production and expression of TAS1, which is the gene involved in TeA production. Results showed that in the field, the summer season presented the highest TeA average level in the grains (587.8 µg/kg) compared to level found in the autumn (440.5 µg/kg). The ecophysiological investigation confirmed that E. sorghinum produces more actively TeA under environmental conditions simulating the summer season. Optimum growth, maximum TAS1 gene expression, and higher extracellular enzymatic production were observed at 26 °C with a water activity of 0.99. Pearson correlation analyses showed that the production of TeA highly correlates with fungal growth. The present study demonstrates that abiotic factors in a combined approach of field and laboratory conditions will assist in predicting the driving environmental factors that could affect growth of E. sorghinum and TeA production in sorghum grains.

Chronic toxicity of graphene and graphene oxide in sequencing batch bioreactors: A comparative investigation.

The present study investigates the chronic toxicity of graphene (G) and graphene oxide (GO) in activated sludge. Sequencing batch bioreactors were fed with influents containing 0, 1 and 5mgL-1 of GO or G (12h cycles) for ten days. Reduction in performance of the bioreactors in relation to chemical oxygen demand, ammonia and phosphate removals was observed after three days in the bioreactors fed with 5mgL-1 of nanomaterials. After about eight days, these reactors reached a steady state nutrient removal, which corresponded to recovery of certain groups of ammonia oxidizing bacteria and phosphate accumulating bacteria despite the increasing accumulation of nanomaterials in the sludge. These results suggested that biological treatment can be affected transiently by initial exposure to the nanomaterials, but certain groups of microorganisms, less sensitive to these nanomaterials, can potentially strive in the presence of these nanomaterials. Results of 16S rRNA gene deep sequencing showed that G and GO affected differently the microbial communities in the activated sludge. Between the two nanomaterials investigated, GO presented the highest impact in nutrient removal, gene abundance and changes in microbial population structures.

Level of Fimbriation Alters the Adhesion of Escherichia coli Bacteria to Interfaces.

Adhesion of bacteria to interfaces is the first step in pathogenic infection, in biofilm formation, and in bioremediation of oil spills and other pollutants. Bacteria use a variety of surface structures to promote interfacial adhesion, with the level of expression of these structures varying in response to local conditions and environmental signals. Here, we investigated how overexpression of type 1 fimbriae, one such appendage, modifies the ability of Escherichia coli to adhere to solid substrates, via biofilm formation and yeast agglomeration, and to oil/water interfaces, via a microbial adhesion to hydrocarbon assay. A plasmid that enables inducible expression of E. coli MG1655 type 1 fimbriae was transformed into fimbriae-deficient mutant strain MG1655ΔfimA. The level of fimH gene expression in the engineered strain, measured using quantitative real-time PCR, could be tuned by changing the concentration of inducer isopropyl ß-d-1-thiogalactopyranoside (IPTG), and was higher than that in strain MG1655. Increasing the degree of fimbriation only slightly modified the surface energy and zeta potential of the bacteria, but enhanced their ability to agglomerate yeast cells and to adhere to solid substrates (as measured by biofilm formation) and to oil/water interfaces. We anticipate that the tunable extent of fimbriation accessible with this engineered strain can be used to investigate how adhesin expression modifies the ability of bacteria to adhere to interfaces and to actively self-assemble there.

Designing polymeric adhesives for antimicrobial materials: poly(ethylene imine) polymer, graphene, graphene oxide and molybdenum trioxide - a biomimetic approach.

The synthesis of biocompatible polymers for coating applications has gained significant attention in recent years due to the increasing spread of infectious diseases via contaminated surfaces. One strategy to combat this problem is to apply antimicrobial coatings to surfaces prone to microbial contamination. This study presents a series of biomimetic polymers that can be used as adhesives to immobilize known antimicrobial agents on the surfaces as coatings. Several polymers containing dopamine methacrylate as co-polymers were synthesized and investigated as adhesives for the deposition of an antimicrobial polymer (polyethyleneimine) and antimicrobial nanoparticles (graphene, graphene oxide and molybdenum trioxide) onto glass surfaces. The results showed that different antimicrobials required different types of adhesives for effective coating. Overall, the coatings fabricated from these composites were shown to inactivate E. coli and B. subtilis within 1 h. These coatings were also effective to prevent biofilm growth and demonstrated to be non-toxic to the human corneal epithelial cell line (htCEpi). Leaching tests of the coatings proved that the coatings were stable under biological conditions.

CO2 sequestration by ureolytic microbial consortia through microbially-induced calcite precipitation.

Urea is an abundant nitrogen-containing compound found in urine of mammals and widely used in fertilizers. This compound is part of the nitrogen biogeochemical cycle and is easily biodegraded by ureolytic microorganisms that have the urease enzyme. Previous studies, with ureolytic isolates, have shown that some ureolytic microorganisms are able to sequester CO2 through a process called microbially-induced calcium carbonate precipitation. The present study investigates 15 ureolytic consortia obtained from the "Pamukkale travertines" and the "Cave Without A Name" using different growth media to identify the possible bacterial genera responsible for CO2 sequestration through the microbially-induced calcite precipitation (MICP). The community structure and diversity were determined by deep-sequencing. The results showed that all consortia presented varying CO2 sequestration capabilities and MICP rates. The CO2 sequestration varied between 0 and 86.4%, and it depended largely on the community structure, as well as on pH. Consortia with predominance of Comamonas, Plesiomonas and Oxalobacter presented reduced CO2 sequestration. On the other hand, consortia dominated by Sporosarcina, Sphingobacterium, Stenotrophomonas, Acinetobacter, and Elizabethkingia showed higher rates of CO2 uptake in the serum bottle headspace.