RESUMO
BACKGROUND: Network inference is an important aim of systems biology. It enables the transformation of OMICs datasets into biological knowledge. It consists of reverse engineering gene regulatory networks from OMICs data, such as RNAseq or mass spectrometry-based proteomics data, through computational methods. This approach allows to identify signalling pathways involved in specific biological functions. The ability to infer causality in gene regulatory networks, in addition to correlation, is crucial for several modelling approaches and allows targeted control in biotechnology applications. METHODS: We performed simulations according to the approximate Bayesian computation method, where the core model consisted of a steady-state simulation algorithm used to study gene regulatory networks in systems for which a limited level of details is available. The simulations outcome was compared to experimentally measured transcriptomics and proteomics data through approximate Bayesian computation. RESULTS: The structure of small gene regulatory networks responsible for the regulation of biological functions involved in biomining were inferred from multi OMICs data of mixed bacterial cultures. Several causal inter- and intraspecies interactions were inferred between genes coding for proteins involved in the biomining process, such as heavy metal transport, DNA damage, replication and repair, and membrane biogenesis. The method also provided indications for the role of several uncharacterized proteins by the inferred connection in their network context. CONCLUSIONS: The combination of fast algorithms with high-performance computing allowed the simulation of a multitude of gene regulatory networks and their comparison to experimentally measured OMICs data through approximate Bayesian computation, enabling the probabilistic inference of causality in gene regulatory networks of a multispecies bacterial system involved in biomining without need of single-cell or multiple perturbation experiments. This information can be used to influence biological functions and control specific processes in biotechnology applications.
Assuntos
Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Redes Reguladoras de Genes , Proteômica , Algoritmos , Bactérias/genética , Teorema de Bayes , Biologia Computacional/métodos , Simulação por Computador , Transdução de Sinais , Biologia de Sistemas/métodosRESUMO
Industrial biomining processes are currently focused on metal sulfides and their dissolution, which is catalyzed by acidophilic iron(II)- and/or sulfur-oxidizing microorganisms. Cell attachment on metal sulfides is important for this process. Biofilm formation is necessary for seeding and persistence of the active microbial community in industrial biomining heaps and tank reactors, and it enhances metal release. In this study, we used a method for direct quantification of the mineral-attached cell population on pyrite or chalcopyrite particles in bioleaching experiments by coupling high-throughput, automated epifluorescence microscopy imaging of mineral particles with algorithms for image analysis and cell quantification, thus avoiding human bias in cell counting. The method was validated by quantifying cell attachment on pyrite and chalcopyrite surfaces with axenic cultures of Acidithiobacillus caldus, Leptospirillum ferriphilum, and Sulfobacillus thermosulfidooxidans. The method confirmed the high affinity of L. ferriphilum cells to colonize pyrite and chalcopyrite surfaces and indicated that biofilm dispersal occurs in mature pyrite batch cultures of this species. Deep neural networks were also applied to analyze biofilms of different microbial consortia. Recent analysis of the L. ferriphilum genome revealed the presence of a diffusible soluble factor (DSF) family quorum sensing system. The respective signal compounds are known as biofilm dispersal agents. Biofilm dispersal was confirmed to occur in batch cultures of L. ferriphilum and S. thermosulfidooxidans upon the addition of DSF family signal compounds.IMPORTANCE The presented method for the assessment of mineral colonization allows accurate relative comparisons of the microbial colonization of metal sulfide concentrate particles in a time-resolved manner. Quantitative assessment of the mineral colonization development is important for the compilation of improved mathematical models for metal sulfide dissolution. In addition, deep-learning algorithms proved that axenic or mixed cultures of the three species exhibited characteristic biofilm patterns and predicted the biofilm species composition. The method may be extended to the assessment of microbial colonization on other solid particles and may serve in the optimization of bioleaching processes in laboratory scale experiments with industrially relevant metal sulfide concentrates. Furthermore, the method was used to demonstrate that DSF quorum sensing signals directly influence colonization and dissolution of metal sulfides by mineral-oxidizing bacteria, such as L. ferriphilum and S. thermosulfidooxidans.
Assuntos
Automação Laboratorial/métodos , Bactérias/metabolismo , Aderência Bacteriana , Metais/metabolismo , Microscopia/métodos , Sulfetos/metabolismo , Acidithiobacillus/metabolismo , Algoritmos , Automação Laboratorial/instrumentação , Biofilmes/crescimento & desenvolvimento , Cobre/metabolismo , Ferro/metabolismo , Consórcios Microbianos , Enxofre/metabolismoRESUMO
A mathematical model of dental plaque has been developed in order to investigate the processes leading to dental caries. The one-dimensional time-dependent model integrates existing knowledge on biofilm processes (mass transfer, microbial composition, microbial conversions and substrate availability) with tooth demineralisation kinetics. This work is based on the pioneering studies of Dibdin who, nearly two decades ago, build a mathematical model roughly describing the metabolic processes taking place in dental plaque. We extended Dibdin's model with: multiple microbial species (aciduric and non-aciduric Streptococci, Actinomyces and Veillonella), more metabolic processes (i.e., aerobic and anaerobic glucose conversion, low and high glucose uptake affinity pathways, formation and consumption of storage compounds), ion transport by Nernst-Planck equations, and we coupled the obtained pH and chemical component gradients inside the plaque with tooth demineralisation. The new model implementation was complemented with faster and more rigorous numerical methods for the model solution. Model results confirm the protective effect of Veillonella due to lactate consumption. Interestingly, on short term, the storage compounds may not necessarily have a negative effect on demineralisation. Individual feeding patterns can also be easily studied with this model. For example, slow ("social") consumption of sugar-containing drinks proves to be more harmful than drinking the same amount over a short period of time.
Assuntos
Placa Dentária/patologia , Modelos Biológicos , Desmineralização do Dente/patologia , Ácidos/metabolismo , Aerobiose , Anaerobiose , Bactérias/metabolismo , Fenômenos Biológicos , Esmalte Dentário/metabolismo , Esmalte Dentário/microbiologia , Placa Dentária/microbiologia , Comportamento de Ingestão de Líquido , Glucose/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Cinética , Fosfatos/metabolismo , Saliva/metabolismo , Fatores de Tempo , Desmineralização do Dente/microbiologiaRESUMO
Society's demand for metals is ever increasing while stocks of high-grade minerals are being depleted. Biomining, for example of chalcopyrite for copper recovery, is a more sustainable biotechnological process that exploits the capacity of acidophilic microbes to catalyze solid metal sulfide dissolution to soluble metal sulfates. A key early stage in biomining is cell attachment and biofilm formation on the mineral surface that results in elevated mineral oxidation rates. Industrial biomining of chalcopyrite is typically carried out in large scale heaps that suffer from the downsides of slow and poor metal recoveries. In an effort to mitigate these drawbacks, this study investigated planktonic and biofilm cells of acidophilic (optimal growth pH < 3) biomining bacteria. RNA and proteins were extracted, and high throughput "omics" performed from a total of 80 biomining experiments. In addition, micrographs of biofilm formation on the chalcopyrite mineral surface over time were generated from eight separate experiments. The dataset generated in this project will be of great use to microbiologists, biotechnologists, and industrial researchers.
Assuntos
Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Metais/isolamento & purificação , Biologia de Sistemas , Ácidos/química , Proteínas de Bactérias/genética , Cobre/isolamento & purificação , RNA Bacteriano/genéticaRESUMO
BACKGROUND: Deep neural networks have been successfully applied to diverse fields of computer vision. However, they only outperform human capacities in a few cases. METHODS: The ability of deep neural networks versus human experts to classify microscopy images was tested on biofilm colonization patterns formed on sulfide minerals composed of up to three different bioleaching bacterial species attached to chalcopyrite sample particles. RESULTS: A low number of microscopy images per category (<600) was sufficient for highly efficient computational analysis of the biofilm's bacterial composition. The use of deep neural networks reached an accuracy of classification of â¼90% compared to â¼50% for human experts. CONCLUSIONS: Deep neural networks outperform human experts' capacity in characterizing bacterial biofilm composition involved in the degradation of chalcopyrite. This approach provides an alternative to standard, time-consuming biochemical methods.