Shedding light on biogas: a transparent reactor triggers the development of a biofilm dominated by Rhodopseudomonas faecalis that holds potential for improved biogas production

AbstractConventional anaerobic digesters intended for the production of biogas usually operate in complete darkness. Therefore, little is known about the effect of light on microbial communities operating in anaerobic digesters. In the present work, we have studied through 16S rRNA gene amplicon Nanopore sequencing and shotgun metagenomic sequencing the taxonomic and functional structure of the microbial community forming a biofilm on the inner wall of a lab-scale transparent anaerobic biodigester illuminated with natural sunlight. The biofilm was composed of microorganisms involved in the four metabolic processes needed for biogas production. The biofilm proved surprisingly rich in Rhodops…

Chemically Stressed Bacterial Communities in Anaerobic Digesters Exhibit Resilience and Ecological Flexibility

Anaerobic digestion is a technology known for its potential in terms of methane production. During the digestion process, multiple metabolites of high value are synthesized. However, recent works have demonstrated the high robustness and resilience of the involved microbiomes; these attributes make it difficult to manipulate them in such a way that a specific metabolite is predominantly produced. Therefore, an exact understanding of the manipulability of anaerobic microbiomes may open up a treasure box for bio-based industries. In the present work, the effect of nalidixic acid, γ-aminobutyric acid (GABA), and sodium phosphate on the microbiome of digested sewage sludge from a water treatmen…

Synthetic microbiology as a source of new enterprises and job creation: a Mediterranean perspective

Peer reviewed

Are multi-omics enough?

Multi-omic techniques are often seen as the future of microbiome studies. We argue that recent strategies for simplifying complex omic-derived data will need to be combined with improved cultivation techniques to pave the way towards a more targeted approach for understanding microbial communities.

The Generalist Inside the Specialist: Gut Bacterial Communities of Two Insect Species Feeding on Toxic Plants Are Dominated by Enterococcus sp.

Some specialist insects feed on plants rich in secondary compounds, which pose a major selective pressure on both the phytophagous and the gut microbiota. However, microbial communities of toxic plant feeders are still poorly characterized. Here, we show the bacterial communities of the gut of two specialized Lepidoptera, Hyles euphorbiae and Brithys crini, which exclusively feed on latex-rich Euphorbia sp. and alkaloid-rich Pancratium maritimum, respectively. A metagenomic analysis based on high-throughput sequencing of the 16S rRNA gene revealed that the gut microbiota of both insects is dominated by the phylum Firmicutes, and especially by the common gut inhabitant Enterococcus sp. Staph…

Liquid co-substrates repower sewage microbiomes

AbstractA range of parameters are known to shape the methanogenic communities of biogas-producing digesters and to strongly influence the amount of biogas produced. In this work, liquid and solid fractions of grass biomass were used separately for semicontinuous batch methanation using sewage sludge as seed sludge. During 6 months of incubation, the amount of input COD was increased gradually, and the underlying methanogenic microbiome was assessed by means of microscopy-based automated cell counting and full-length 16S rRNA high-throughput sequencing. In this sense, we prove for the first time the suitability of the ONT™MinION platform as a monitoring tool for anaerobic digestion systems. …

Selecting Microbial Strains from Pine Tree Resin: Biotechnological Applications from a Terpene World

Resin is a chemical and physical defensive barrier secreted by many plants, especially coniferous trees, with insecticidal and antimicrobial properties. The degradation of terpenes, the main components accounting for the toxicity of resin, is highly relevant for a vast range of biotechnological processes, including bioremediation. In the present work, we used a resin-based selective medium in order to study the resin-tolerant microbial communities associated with the galls formed by the moth Retinia resinella; as well as resin from Pinus sylvestris forests, one of the largest ecosystems on Earth and a yet-unexplored source of terpene-degrading microorganisms. The taxonomic and functional di…

The coffee-machine bacteriome: biodiversity and colonisation of the wasted coffee tray leach

AbstractMicrobial communities are ubiquitous in both natural and artificial environments. However, microbial diversity is usually reduced under strong selection pressures, such as those present in habitats rich in recalcitrant or toxic compounds displaying antimicrobial properties. Caffeine is a natural alkaloid present in coffee, tea and soft drinks with well-known antibacterial properties. Here we present the first systematic analysis of coffee machine-associated bacteria. We sampled the coffee waste reservoir of ten different Nespresso machines and conducted a dynamic monitoring of the colonization process in a new machine. Our results reveal the existence of a varied bacterial community…

Assembly methods for nanopore-based metagenomic sequencing: a comparative study

ABSTRACTBackgroundMetagenomic sequencing has lead to the recovery of previously unexplored microbial genomes. In this sense, short-reads sequencing platforms often result in highly fragmented metagenomes, thus complicating downstream analyses. Third generation sequencing technologies, such as MinION, could lead to more contiguous assemblies due to their ability to generate long reads. Nevertheless, there is a lack of studies evaluating the suitability of the available assembly tools for this new type of data.FindingsWe benchmarked the ability of different short-reads and long-reads tools to assembly two different commercially available mock communities, and observed remarkable differences i…

1H,13C and19F NMR spectroscopy of polyfluorinated ureas. Correlations involving NMR chemical shifts and electronic substituent effects

Seventeen N-(mono-, di-, tri-, tetra- and penta-fluorophenyl)-N′-(3-nitrophenyl)ureas were prepared and characterized. Complete assignment of their 1H, 13C and 19F NMR data was undertaken and the correlation of the chemical shifts of the ureido protons with field-inductive and mesomeric electronic substituent parameters was studied using the Swain–Lupton model. The best correlations were obtained when the study was limited to certain substitution patterns, e.g. non-ortho, mono-ortho- and di-ortho-fluorinated ureas, which reveal probable changes in conformations caused by the degree of ortho fluorination at the phenyl ring. Additionally, there is an excellent linear cross-correlation between…

Ammonia removal during leach-bed acidification leads to optimized organic acid production from chicken manure

This work demonstrates the suitability of nitrogen removal during anaerobic acidification in batch configuration for a more efficient pre-treatment of chicken manure prior to anaerobic digestion. High loading rates corresponding to a total nitrogen input between 6.3 and 9.5 g L−1 allowed successful suppression of methanogenic archaea. To eliminate nitrogen, NH3-stripping and MAP (magnesium ammonium phosphate hexahydrate) precipitation were compared. In spite of decreased cell quantities detected using qPCR, removal of nitrogen caused an increase in volatile fatty acid (VFA) formation from 13 to 19%. The highest nitrogen removal during acidification (up to 29%) was achieved with three consec…

Unveiling Bacterial Interactions through Multidimensional Scaling and Dynamics Modeling

AbstractWe propose a new strategy to identify and visualize bacterial consortia by conducting replicated culturing of environmental samples coupled with high-throughput sequencing and multidimensional scaling analysis, followed by identification of bacteria-bacteria correlations and interactions. We conducted a proof of concept assay with pine-tree resin-based media in ten replicates, which allowed detecting and visualizing dynamical bacterial associations in the form of statistically significant and yet biologically relevant bacterial consortia.

Preparation and Promotion of Fruit Growth in Kiwifruit of Fluorinated N-Phenyl-N‘-1,2,3-thiadiazol-5-yl Ureas

Seventeen phenyl-fluorinated analogues of thidiazuron [N-phenyl-N'-(1,2,3-thiadiazol-5-yl)urea, TDZ] have been prepared and characterized. The effects of each fluorinated urea on growth and quality of kiwifruits (Actinidia deliciosa) were evaluated by comparison with untreated (control) and TDZ-treated fruits. The results obtained showed a clear dependence of the growth-promoting activity of these fluorinated ureas on the pattern and degree of fluorine substitution in the phenyl ring. The most effective for promoting fruit growth was N-(2,3,5,6-tetrafluorophenyl)-N'-(1',2',3'-thiadiazol-5'-yl)urea at 25 ppm (at harvest, treated fruits were 58% heavier than untreated ones) followed by N-(3,5…

Shedding light on biogas: Phototrophic biofilms in anaerobic digesters hold potential for improved biogas production

Conventional anaerobic digesters intended for the production of biogas usually operate in complete darkness. Therefore, little is known about the effect of light on their microbial communities. In the present work, 16S rRNA gene amplicon Nanopore sequencing and shotgun metagenomic sequencing were used to study the taxonomic and functional structure of the microbial community forming a biofilm on the inner wall of a laboratory-scale transparent anaerobic biodigester illuminated with natural sunlight. The biofilm was composed of microorganisms involved in the four metabolic processes needed for biogas production, and it was surprisingly rich in Rhodopseudomonas faecalis, a versatile bacterium…

Sphingomonas solaris sp. nov., isolated from a solar panel in Boston, Massachusetts

Solar panel surfaces, although subjected to a range of extreme environmental conditions, are inhabited by a diverse microbial community adapted to solar radiation, desiccation and temperature fluctuations. This is the first time a new bacterial species has been isolated from this environment. Strain R4DWNT belongs to the genus Sphingomonas and was isolated from a solar panel surface in Boston, MA, USA. Strain R4DWNT is a Gram-negative, non-motile and rod-shaped bacteria that tested positive for oxidase and catalase and forms round-shaped, shiny and orange-coloured colonies. It is mesophilic, neutrophilic and non-halophilic, and presents a more stenotrophic metabolism than its closest neighb…

Extremophilic microbial communities on photovoltaic panel surfaces: a two‐year study

Solar panel surfaces can be colonized by microorganisms adapted to desiccation, temperature fluctuations and solar radiation. Although the taxonomic and functional composition of these communities has been studied, the microbial colonization process remains unclear. In the present work, we have monitored this microbial colonization process during 24 months by performing weekly measurements of the photovoltaic efficiency, carrying out 16S rRNA gene high‐throughput sequencing, and studying the effect of antimicrobial compounds on the composition of the microbial biocenosis. This is the first time a long‐term study of the colonization process of solar panels has been performed, and our results…

Additional file 1: Table S1. of Standards not that standard

Biobrick parts (Bb1 to Bb6) included in the different DNA constructions tested in this work. Table S2. Proteins displaying statistically significant alterations in their expression levels, as detected by iTRAQ analysis (DOCX 18 kb).

Standards not that standard

There is a general assent on the key role of standards in Synthetic Biology. In two consecutive letters to this journal, suggestions on the assembly methods for the Registry of standard biological parts have been described. We fully agree with those authors on the need of a more flexible building strategy and we highlight in the present work two major functional challenges standardization efforts have to deal with: the need of both universal and orthogonal behaviors. We provide experimental data that clearly indicate that such engineering requirements should not be taken for granted in Synthetic Biology. Electronic supplementary material The online version of this article (doi:10.1186/s1303…

A highly diverse, desert-like microbial biocenosis on solar panels in a Mediterranean city

AbstractMicroorganisms colonize a wide range of natural and artificial environments although there are hardly any data on the microbial ecology of one the most widespread man-made extreme structures: solar panels. Here we show that solar panels in a Mediterranean city (Valencia, Spain) harbor a highly diverse microbial community with more than 500 different species per panel, most of which belong to drought-, heat- and radiation-adapted bacterial genera, and sun-irradiation adapted epiphytic fungi. The taxonomic and functional profiles of this microbial community and the characterization of selected culturable bacteria reveal the existence of a diverse mesophilic microbial community on the …

Eubacteria and archaea communities in seven mesophile anaerobic digester plants in Germany

Background Only a fraction of the microbial species used for anaerobic digestion in biogas production plants are methanogenic archaea. We have analyzed the taxonomic profiles of eubacteria and archaea, a set of chemical key parameters, and biogas production in samples from nine production plants in seven facilities in Thuringia, Germany, including co-digesters, leach-bed, and sewage sludge treatment plants. Reactors were sampled twice, at a 1-week interval, and three biological replicates were taken in each case. Results A complex taxonomic composition was found for both eubacteria and archaea, both of which strongly correlated with digester type. Plant-degrading Firmicutes as well as Bacte…

Towards a microbial thermoelectric cell.

Microbial growth is an exothermic process. Biotechnological industries produce large amounts of heat, usually considered an undesirable by-product. In this work, we report the construction and characterization of the first microbial thermoelectric cell (MTC), in which the metabolic heat produced by a thermally insulated microbial culture is partially converted into electricity through a thermoelectric device optimized for low ΔT values. A temperature of 41°C and net electric voltage of around 250–600 mV was achieved with 1.7 L baker’s yeast culture. This is the first time microbial metabolic energy has been converted into electricity with an ad hoc thermoelectric device. These results might…

Aequorin-expressing yeast emits light under electric control

In this study, we show the use of direct external electrical stimulation of a jellyfish luminescent calcium-activated protein, aequorin, expressed in a transgenic yeast strain. Yeast cultures were electrically stimulated through two electrodes coupled to a standard power generator. Even low (1.5. V) electric pulses triggered a rapid light peak and serial light pulses were obtained after electric pulses were applied periodically, suggesting that the system is re-enacted after a short refraction time. These results open up a new scenario, in the very interphase between synthetic biology and cybernetics, in which complex cellular behavior might be subjected to electrical control.

A lab in the field: applications of real-time, in situ metagenomic sequencing

High-throughput metagenomic sequencing is considered one of the main technologies fostering the development of microbial ecology. Widely used second-generation sequencers have enabled the analysis of extremely diverse microbial communities, the discovery of novel gene functions, and the comprehension of the metabolic interconnections established among microbial consortia. However, the high cost of the sequencers and the complexity of library preparation and sequencing protocols still hamper the application of metagenomic sequencing in a vast range of real-life applications. In this context, the emergence of portable, third-generation sequencers is becoming a popular alternative for the rapi…

Art‐omics: multi‐omics meet archaeology and art conservation

Summary Multi‐omics can informally be described as the combined use of high‐throughput techniques allowing the characterization of complete microbial communities by the sequencing/identification of total pools of biomolecules including DNA, proteins or metabolites. These techniques have allowed an unprecedented level of knowledge on complex microbial ecosystems, which is having key implications in land and marine ecology, industrial biotechnology or biomedicine. Multi‐omics have recently been applied to artistic or archaeological objects, with the goal of either contributing to shedding light on the original context of the pieces and/or to inform conservation approaches. In this minireview,…

Engineering Bacteria to Form a Biofilm and Induce Clumping in Caenorhabditis elegans

Bacteria are needed for a vast range of biotechnological processes, which they carry out either as pure cultures or in association with other bacteria and/or fungi. The potential of bacteria as biofactories is hampered, though, by their limited mobility in solid or semisolid media such as agricultural or domestic waste. This work represents an attempt toward overcoming this limitation by associating bacterial biotechnological properties with the transport ability of the nematode Caenorhabditis elegans. We report here biofilm formation on C. elegans by engineered Escherichia coli expressing a Xhenorhabdus nematophila adhesion operon and induction of nematode social feeding behavior (clumping…

Thermoelectric Heat Exchange and Growth Regulation in a Continuous Yeast Culture

We have designed a thermoelectric heat exchanger (TEHE) for microbial fermentations that is able to produce electric power from a microbial continuous culture using the intrinsic heat generated by microbial growth. While the TEHE was connected, the system proved able to stably self-maintain both the temperature and the optical density of the culture. This paves the way toward a more sustainable operation of microbial fermentations, in which energy could be saved by converting part of the metabolic heat into usable electric power.

A round trip to the desert: In situ nanopore sequencing informs targeted bioprospecting

Bioprospecting expeditions are often performed in remote locations, in order to access previously unexplored samples. Nevertheless, the actual potential of those samples is only assessed once scientists are back in the laboratory, where a time-consuming screening must take place. This work evaluates the suitability of using Nanopore sequencing during a journey to the Tabernas Desert (Spain) for forecasting the potential of specific samples in terms of bacterial diversity and prevalence of radiation- and desiccation-resistant taxa, which were the target of the bioprospecting activities. Samples collected during the first day were analyzed through 16S rRNA gene sequencing using a mobile labor…

Bioprospecting challenges in unusual environments

Editorial: The microbiome as a source of new enterprises and job creation.

Towards light-mediated sensing of bacterial comfort

Abstract Bacterial comfort is central to biotechnological applications. Here, we report the characterization of different sensoring systems, the first step within a broader synthetic biology-inspired light-mediated strategy to determine Escherichia coli perception of environmental factors critical to bacterial performance. We did so by directly ‘asking’ bacterial cultures with light-encoded questions corresponding to the excitation wavelength of fluorescent proteins placed under the control of environment-sensitive promoters. We built four genetic constructions with fluorescent proteins responding to glucose, temperature, oxygen and nitrogen; and a fifth construction allowing UV-induced exp…

iGEM 2.0—refoundations for engineering biology

A Bacillus thuringiensis strain producing epizootics on Plodia interpunctella: A case study

Abstract After several disease outbreaks in laboratory cultures of pyralid moths in Tabriz University, Iran, during 2004 and 2005, a new Bacillus thuringiensis aizawai strain EF495116 (BTA) was isolated from a dead Plodia interpunctella larva. A complete characterization of the strain was performed, including serological identification, protein and plasmid pattern determination, a PCR-based identification of virulence-related genes, nucleotide sequence analysis of the 16S rDNA and gyrB genes (in order to find out relationships between the species with other virulent Bacillus pathogens), and biological activity assays. These studies revealed that BTA produced a major parasporal protein band …

Bacteria from acidic to strongly alkaline insect midguts: Potential sources of extreme cellulolytic enzymes

Abstract Bacterial isolates from the European Corn Borer (ECB) Ostrinia nubilalis and the coleopteran Colorado Potato Beetle (CPB) Leptinotarsa decemlineata midguts were identified and characterized. Twenty-four colonies, selected on solid media with cellulose as the sole carbon source and pH values ranging from 5 to 12, were identified through 16S sequencing as members of the genera Acinetobacter , Comamonas , Klebsiella , Microbacterium , Micrococcus and Sphingobacterium . A complete enzymatic characterization revealed widespread – albeit moderate – cellulase properties in all but one isolate and high xylanase activity in the four CPB isolates. Different enzymatic patterns in terms of opt…