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At least 217 records · Page 12

Syntrophic bacterial and host–microbe interactions in bacterial vaginosis

Bacterial vaginosis (BV) is a common, polymicrobial condition of the vaginal microbiota that is associated with symptoms such as malodor and excessive discharge, along with increased risk of various adverse sequelae. Host–bacteria and bacteria–bacteria interactions are thought to contribute to the condition, but many of these functions have yet to be elucidated. Using untargeted metaproteomics, we identified 1068 host and 1418 bacterial proteins in a set of cervicovaginal lavage samples collected from 20 participants with BV and 9 who were negative for the condition. We identified Dialister micraerophilus as a major producer of malodorous polyamines and identified a syntrophic interaction between this organism and Fannyhessea vaginae that leads to increased production of putrescine, a metabolite characteristic of BV. Although formate synthesis has not previously been noted in BV, we discovered diverse bacteria associated with the condition express pyruvate formate-lyase enzymes in vivo and confirm these organisms secrete formic acid in vitro. Sodium hypophosphite efficiently inhibited this function in multiple taxa. We also found that the fastidious organism Coriobacteriales bacterium DNF00809 can metabolize formic acid secreted by Gardnerella vaginalis, representing another syntrophic interaction. We noted an increased abundance of the host epithelial repair protein transglutaminase 3 in the metaproteomic data, which we confirmed by enzyme-linked immunosorbent assay. Other proteins identified in our samples implicate Finegoldia magna and Parvimonas micra in the production of malodorous trimethylamine. Some bacterial proteins identified represent novel targets for future therapeutics to disrupt BV communities and promote vaginal colonization by commensal lactobacilli.

bacterial vaginosis↗

Evaluation of Protein Extraction Methods for Metaproteomic Analyses of Root-Associated Microbes

Metaproteomics is a powerful tool for the characterization of metabolism, physiology, and functional interactions in microbial communities, including plant-associated microbiota. However, the metaproteomic methods that have been used to study plant-associated microbiota are very laborious and require large amounts of plant tissue, hindering wider application of these methods. We optimized and evaluated different protein extraction methods for metaproteomics of plant-associated microbiota in two different plant species (Arabidopsis and maize). Our main goal was to identify a method that would work with low amounts of input material (40 to 70 mg) and that would maximize the number of identified microbial proteins. We tested eight protocols, each comprising a different combination of physical lysis method, extraction buffer, and cell-enrichment method on roots from plants grown with synthetic microbial communities. We assessed the performance of the extraction protocols by liquid chromatography-tandem mass spectrometry–based metaproteomics and found that the optimal extraction method differed between the two species. For Arabidopsis roots, protein extraction by beating whole roots with small beads provided the greatest number of identified microbial proteins and improved the identification of proteins from gram-positive bacteria. For maize, vortexing root pieces in the presence of large glass beads yielded the greatest number of microbial proteins identified. Based on these data, we recommend the use of these two methods for metaproteomics with Arabidopsis and maize. Furthermore, detailed descriptions of the eight tested protocols will enable future optimization of protein extraction for metaproteomics in other dicot and monocot plants.

59 BASIC BIOLOGICAL SCIENCES↗

The ‘porin–cytochrome’ model for microbe-to-mineral electron transfer

Many species of bacteria can couple anaerobic growth to the respiratory reduction of insoluble minerals containing Fe(III) or Mn(III/IV). It has been suggested that in Shewanella species electrons cross the outer membrane to extracellular substrates via 'porin-cytochrome' electron transport modules. The molecular structure of an outer-membrane extracellular-facing deca-haem terminus for such a module has recently been resolved. It is debated how, once outside the cells, electrons are transferred from outer-membrane cytochromes to insoluble electron sinks. This may occur directly or by assemblies of cytochromes, perhaps functioning as 'nanowires', or via electron shuttles. Here we review recent work in this field and explore whether it allows for unification of the electron transport mechanisms supporting extracellular mineral respiration in Shewanella that may extend into other genera of Gram-negative bacteria.

59 BASIC BIOLOGICAL SCIENCES↗

TES: Modelling Microbes to Predict Post-Fire Carbon Cycling in the Boreal Forest across Burn Severities (Exploratory Proposal)

Wildfires in boreal ecosystems represent a globally significant ecological process that is expected to be sensitive to changing climate as well as forest management strategies, but is currently insufficiently understood and represented in models. We sought to determine whether linking belowground microbial community composition, size, and activity to aboveground properties of burn severity and plant community composition (upland jack pine and upland spruce) would allow us to better model post-fire soil CO 2 fluxes using the Carbon, Organisms, Rhizosphere and Protection in the Soil Environment (CORPSE) model. We used an integrated modelling experimental approach, with mechanistic laboratory experiments designed to inform models.

54 ENVIRONMENTAL SCIENCES↗

Microbes Share Rides Too: Updating Encapsulated Bioburden Estimate Values in Electronic Parts Common to Class D Mission Platforms.

As NASA develops more planetary protection missions that are Category III and in the Class D/rideshare mission platform, there is an increasing reliance on estimation of prelaunch bioburden to understand needs for burn up and breakup analyses or other methods to meet pre-launch bioburden levels. Current levels estimated for the encapsulated bioburden of semiconductor parts cited by NASA have been based on estimates of semiconductor manufacturing approaches from the 1970s that do not incorporate over fifty years of evolving cleanliness of semiconductor manufacturing processes and facilities. This investigation will focus on direct sampling of semiconductor parts for encapsulated bioburden values, drawing upon Goddard’s strength in EEE parts, supply chain management, destructive parts analysis and existing local planetary protection lab facilities to prepare and measure encapsulated bioburden at a statistically significant level for standard electronics parts common to known Class D/rideshare platforms.

planetary protection↗

Microbes Share Rides Too: Updating Encapsulated Bioburden Estimate Values in Electronic Parts Common to Class D Missions

As NASA develops more planetary protection missions that are Category III and in the Class D/rideshare mission platform, there is an increasing reliance on estimation of prelaunch bioburden to understand needs for burn up and breakup analyses or other methods to meet pre-launch bioburden levels. Current levels estimated for the encapsulated bioburden of semiconductor parts cited by NASA have been based on estimates of semiconductor manufacturing approaches from the 1970s that do not incorporate the evolving cleanliness of semiconductor manufacturing since that time. This investigation will focus on direct sampling of semiconductor parts for encapsulated bioburden values, drawing upon Goddard’s strength in EEE parts, supply chain management, destructive parts analysis and existing local planetary protection lab facilities to prepare and measure encapsulated bioburden at a statistically significant level for standard electronics parts common to known Class D/rideshare platforms.

planetary protection↗

Revealing the Role of Microbes in Controlling Contaminants

In Rifle, Colorado, Berkeley Lab earth scientist, Kenneth Hurst Williams, highlights the role subsurface microbial communities can play in controlling the flow of contaminants in groundwater. The DOE Joint Genome Institute is a key collaborator in the research. Williams is Component Lead of Watershed Structure and Controls within Berkeley Lab's Genomes-to-Watershed Scientific Focus Area.

54 ENVIRONMENTAL SCIENCES↗

Sink Engineering in Photosynthetic Microbes

Photosynthetic organisms must balance harvested light energy (source) with a corresponding capacity to utilize the energy via downstream metabolic processes (sink). Insufficient source energy leads to starvation conditions, while insufficient sink capacity results in an overreduction of the electron transport chain, reduced photosynthetic efficiency, and photodamage. Despite the central importance of source/sink balancing in photosynthesis, our understanding of the mechanisms used to control this process in remains poor across diverse phototrophs. In particular, these balancing processes are essentially unstudied in cyanobacteria and microalgae, which possess distinct needs relative to the multicellular plants that have served as the model organisms for elucidating source/sink pathways. Recent bioengineering efforts have uncovered phenomena that are consistent with a well-developed source/sink sensing machinery in unicellular phototrophs. Furthermore, a deeper understanding of the fundamental mechanisms underlying source/sink dynamics in cyanobacteria and algae may prove to be critical for our capacity to boost photosynthetic efficiencies in these organisms, and enable their use as alternative crop species.

Carbon Fixation↗

Rhizosphere Carbon Turnover from Cradle to Grave: The Role of Microbe–Plant Interactions

Plant roots are the primary source of organic materials that become stabilized in soil. While most root carbon is decomposed into carbon dioxide (CO 2 ), the remainder typically undergoes multiple microbial transformations before it forms longer-term associations with soil minerals. However, the mechanisms by which roots affect microbial utilization of organic materials and subsequent mineral stabilization processes are poorly understood. It is well known that living roots increase the biomass of nearby microbial communities, and shape their population dynamics, diversity, and interactions. Community assembly and metabolic potential of these rhizosphere-enriched microorganisms are strongly influenced by the chemical composition of the exudates released by the host plant. The root exudate pools of plants undergo compositional changes as they grow, reproduce, and senesce. In the well-studied annual grasses Avena barbata and Avena fatua, this changing rhizosphere substrate pool and the “bloom” of organisms that respond are phylogenetically coherent; Acidobacteria and Actinobacteria are consistently depleted, whereas Alpha and Betaproteobacteria and Bacteroidetes are reliably enriched. When compared to non-root-influenced bulk soils, the responsive community is predictably less taxon-rich, yet forms more complex networks. Finally, these rhizosphere dynamics have significant downstream effects on the colonization of nearby soil minerals, degradation of prior season’s root litters, and the balance of stabilized versus lost soil carbon.

Pett-Ridge, Jennifer↗

Biodegradation of Poly(Aspartic Acid-Itaconic Acid) Copolymers by Miscellaneous Microbes from Natural Water

Poly(aspartic acid-itaconic acid) copolymers (PAI) is a new scale inhibitor for water treatment. Thus, it is necessary to investigate its biodegradability. The biodegradability of PAI was investigated through CO{sub 2} evolution tests under different conditions based on determination of carbon dioxide production. The investigation results showed that the degradation rate of PAI on day 10 and day 28 were respectively 38.7 and 79.5%, indicating that PAI was one kind of easily biodegradable scale inhibitors. With the increase in the content of itaconic acid in copolymerization process, the biodegradability of PAI was significantly reduced. In addition, the high biodegradability might be attributed to the existence of C–N bone-structure and more –COO–. Finally, Cu{sup 2+} could decrease the degradation percentage and the enzyme inhibition effect of Cu{sup 2+} was not the linear effect, but the “low-dosage effect”.

54 ENVIRONMENTAL SCIENCES↗

Advances in monitoring environmental microbes

Culture-independent approaches, such as next-generation sequencing and microarray-based tools, provide insight into the identity and functional diversity of microbial communities. Although these approaches are potentially powerful tools in understanding microbial structure and function, there are a number of limitations that may bias conclusions. Here, in order to mitigate these biases, an understanding of potential biases within each stage of the experimental process is necessary. This review focuses on the biases associated with sample collection, nucleic acid extraction, processing, sequencing analyses, and Chip technologies used in microbial ecology studies.

54 ENVIRONMENTAL SCIENCES↗