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Biofouling has been persisting as a global problem due to the difficulties in finding efficient and environmentally friendly antifouling coatings for long-term applications. Initial attachment of bacteria on material surface and subsequent formation of biofilm are the predominate phenomena accounting for subsequent occurrence of biofouling. Among the various factors influencing the bacterial attachment, conditioning layer formed by organic macromolecules usually plays the key role in mediating bacterial attachment through altering physicochemical properties of substrate surface. In this study, a guanidine-modified polysaccharide conditioning layer with the capability of tuning the bacterial attachment is constructed and characterized. Dextran, a polysaccharide widespread in bacteria extracellular polymeric substances (EPS), is oxidized by sodium periodate, and cationic polymer polyhexamethylene guanidine hydrochloride (PHMG) is anchored to oxidized dextran (ODEX) by Schiff base reaction. AFM characterization reveals morphological changes of the polysaccharide conditioning layer from tangled chain to island conformation after the PHMG modification. The guanidine-based dextran conditioning layer promotes attachment of both P. aeruginosa and S. aureus and disrupted bacterial cytomembranes are seen for the attached bacteria due to electrostatic interaction of the electropositive guanidine group with the electronegative bacteria. The guanidine-based dextran conditioning layer shows a low survival ratio of 22 %-34 % and 1 %-4 % for P. aeruginosa and S. aureus respectively after incubation in the bacterial suspension for 72 hours. The results would give insight into further exploring the potential applications of the newly designed polysaccharides conditioning layer for combating occurrence of biofouling.

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This study aims to present a comprehensive study on the risks associated with the residual presence and transport of Escherichia coli (E. coli) in soil following the application of livestock manure in Chinese farmlands by integrating machine learning algorithms with mechanism-based models (Phydrus). We initially review 28 published papers to gather data on E. coli's die-off and attachment characteristics in soil. Machine learning models, including deep learning and gradient boosting machine, are employed to predict key parameters such as the die-off rate of E. coli and first-order attachment coefficient in soil. Then, Phydrus was used to simulate E. coli transport and survival in 23692 subregions in China. The model considered regional differences in E. coli residual risk and transport, influenced by soil properties, soil depths, precipitation, seasonal variations, and regional disparities. The findings indicate higher residual risks in regions such as the Northeast China, Eastern Qinghai-Tibet Plateau, and pronounced transport risks in the fringe of the Sichuan Basin fringe, the Loess Plateau, the North China Plain, the Northeast Plain, the Shigatse Basin, and the Shangri-La region. The study also demonstrates a significant reduction in both residual and transport risks one month after manure application, highlighting the importance of timing manure application and implementing region-specific standards. This research contributes to the broader understanding of pathogen behavior in agricultural soils and offers practical guidelines for managing the risks associated with manure use. This study's comprehensive method offers a potentially valuable tool for evaluating microbial contaminants in agricultural soils across the globe.

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OBJECTIVES: To create bacteria-resistant dental CAD-CAM blocks with a biofilm-resistant effect by incorporating Nano-crystalline ceramic and polymer (NCP) with 2-methacryloyloxyethyl phosphorylcholine (MPC) and sulfobetaine methacrylate (SBMA) and at an equimolar ratio, referred to as MS. METHODS: Experimental groups comprised NCP blocks containing zwitterions at 0.15wt% (MS015) and 0.45wt% (MS045). NCP blocks without MS served as control (CTRL). Flexural strength, surface hardness, water sorption and solubility, photometric properties, and cytotoxicity were assessed for all samples. Additionally, the resistance to single and multi-species bacterial adhesion was investigated. RESULTS: MS045 showed significant reduction in flexural strength (P < 0.01) compared to both CTRL and MS015. Both MS015 and MS045 showed significantly increased water sorption and significant reduction in water solubility compared to CTRL. Light transmission remained consistent across all MS content levels, but the irradiance value decreased by 12 % in the MS045 group compared to the MS015 group. Notably, compared to the CTRL group, the MS015 group exhibited enhanced resistance to adhesion by Porphyromonas gingivalis and a multi-species salivary biofilm, with biofilm thickness and biomass reduced by 45 % and 56 %, respectively. CONCLUSIONS: NCP containing 0.15 % MS can effectively reduce adhesion of multiple species of bacteria while maintaining physical and mechanical properties. CLINICAL SIGNIFICANCE: NCP integrating zwitterions is clinically advantageous in resisting bacterial adhesion at internal and external margins of milled indirect restoration.

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High-temperature-requirement protein A (HtrA) family proteins play important roles in controlling protein quality and are recognized as virulence factors in numerous animal and human bacterial pathogens. The role of HtrA family proteins in plant pathogens remains largely unexplored. Here, we investigated the HtrA family protein, DegQ, in the crucifer black rot pathogen Xanthomonas campestris pathovar campestris (Xcc). DegQ is essential for bacterial attachment and full virulence of Xcc. Moreover, the degQ mutant strain showed increased sensitivity to heat treatment and sodium dodecyl sulfate. Expressing the intact degQ gene in trans in the degQ mutant could reverse the observed phenotypic changes. In addition, we demonstrated that the DegQ protein exhibited chaperone-like activity. Transcriptional analysis displayed that degQ expression was induced under heat treatment. Our results contribute to understanding the function and expression of DegQ of Xcc for the first time and provide a novel perspective about HtrA family proteins in plant pathogen.

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This study examines the microbial colonization characteristics of microplastics (MPs) in wastewater treatment plants (WWTPs), focusing on polymer types (High-Density Polyethylene (HDPE) and Polyethylene Terephthalate (PET)) and various stages of wastewater treatments. Through individual and sequential deployment approaches, the research aimed to identify the determinants of bacterial colonization on MPs, whether they were introduced at each stage of treatment individually or in sequence from primary to tertiary stages. The study revealed that the stage of wastewater treatment profoundly influenced bacterial colonization on the polymer types MPs, with bacterial attachment being largely niche-specific. HDPE showed increased sensitivity to wastewater composition, leading to selective biofilm formation. For instance, in HDPE, Firmicutes accounted for 25.1 ± 0.04 % during primary treatment, while Alphaproteobacteria increased significantly in the tertiary treatment to 19.8 ± 0.1 %. Conversely, PET exhibited a stochastic pattern of bacterial colonization due to differences in surface hydrophilicity. Additionally, in sequential deployments, a notable shift towards stochastic bacterial attachment on MPs, particularly with HDPE was observed. The Shannon diversity values for MP biofilms were consistently higher than those for wastewater across all stages, with PET showing an increase in diversity in sequential deployments (Shannon diversity: 5.01 ± 0.03 for tertiary stage). These findings highlight the critical role of MPs as carriers of diverse bacteria, emphasizing the necessity for strategies to mitigate their impact in WWTPs. This study presents a significant advancement in our understanding of the interactions between MPs and microbial populations in WWTP environments.

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Quorum Sensing (QS) is a form of cell-to-cell communication that enables bacteria to modify behavior according to their population density. While QS has been proposed as a potential intervention against pathogen infection, QS-mediated communication within the mammalian digestive tract remains understudied. Using an LC-MS/MS approach, we discovered that Citrobacter rodentium, a natural murine pathogen used to model human infection by pathogenic Escherichia coli, utilizes the CroIR system to produce three QS-molecules. We then profiled their accumulation both in vitro and across different gastrointestinal sites over the course of infection. Importantly, we found that in the absence of QS capabilities the virulence of C. rodentium is enhanced. This highlights the role of QS as an effective mechanism to regulate virulence according to the pathogen's spatio-temporal context to optimize colonization and transmission success. These results also demonstrate that inhibiting QS may not always be an effective strategy for the control of virulence.

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Membrane-based wastewater reclamation is used to mitigate water scarcity; however, irreversible biofouling is an elusive problem that hinders the efficiency of a forward-osmosis (FO) membrane-based process, and the protein responsible for fouling is unknown. Herein, we identified fouling proteins by analyzing the microbiome and proteome of wastewater extracellular polymeric substances responsible for strong irreversible FO-membrane fouling. The IGLSSLPR peptide of a PilZ domain-containing protein was found to recruit bacterial attachment when immobilized on the membrane surface while suppressing it when dissolved, in a similar manner to the Arg-Gly-Asp (RGD) peptide in mammalian cell cultures. Bacteria adhere to IGLSSLPR and poly-l-lysine-coated membranes with similar energies and exhibit water fluxes that decline similarly, which is ascribable to interaction as strong as electrostatic interactions in the peptide-coated membranes. We conclude that IGLSSLPR is the key domain responsible for membrane fouling and can be used to develop antifouling technology against bacteria, which is similar to the current usage of RGD peptide in mammalian cell cultures.

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Fresh-cut produces are often consumed uncooked, thus proper sanitation is essential for preventing cross contamination. The reduction and subsequent growth of Salmonella enterica sv Thompson were studied in pre-cut iceberg lettuce washed with simulated wash water (SWW), sodium hypochlorite (SH, free chlorine 25 mg/L), and peroxyacetic acid (PAA, 80 mg/L) and stored for 9 days under modified atmosphere at 9, 13, and 18 °C. Differences in reduction between SH and PAA were non-existent. Overall, visual quality, dehydration, leaf edge and superficial browning and aroma during storage at 9 °C were similar among treatments, but negative effects increased with temperature. These results demonstrated that PAA can be used as an effective alternative to chlorine for the disinfection of Salmonella spp. in fresh-cut lettuce. The growth of Salmonella enterica sv Thompson was successfully described with the Baranyi and Roberts growth model in the studied storage temperature range, and after treatment with SWW, chlorine, and PAA. Subsequently, predictive secondary models were used to describe the relationship between growth rates and temperature based on the models' family described by Belehrádek. Interestingly, the exposure to disinfectants biased growth kinetics of Salmonella during storage. Below 12 °C, growth rates in lettuce treated with disinfectant (0.010-0.011 log CFU/h at 9 °C) were lower than those in lettuce washed with water (0.016 log CFU/h at 9 °C); whereas at higher temperatures, the effect was the opposite. Thus, in this case, the growth rate values registered at 18 °C for lettuce treated with disinfectant were 0.048-0.054 log CFU/h compared to a value of 0.038 log CFU/h for lettuce treated with only water. The data and models developed in this study will be crucial to describing the wash-related dynamics of Salmonella in a risk assessment framework applied to fresh-cut produce, providing more complete and accurate risk estimates.

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Bacterial infections are increasingly problematic due to the rise of antimicrobial resistance. Consequently, the rational design of materials naturally resistant to biofilm formation is an important strategy for preventing medical device-associated infections. Machine learning (ML) is a powerful method to find useful patterns in complex data from a wide range of fields. Recent reports showed how ML can reveal strong relationships between bacterial adhesion and the physicochemical properties of polyacrylate libraries. These studies used robust and predictive nonlinear regression methods that had better quantitative prediction power than linear models. However, as nonlinear models' feature importance is a local rather than global property, these models were hard to interpret and provided limited insight into the molecular details of material-bacteria interactions. Here, we show that the use of interpretable mass spectral molecular ions and chemoinformatic descriptors and a linear binary classification model of attachment of three common nosocomial pathogens to a library of polyacrylates can provide improved guidance for the design of more effective pathogen-resistant coatings. Relevant features from each model were analyzed and correlated with easily interpretable chemoinformatic descriptors to derive a small set of rules that give model features tangible meaning that elucidate relationships between the structure and function. The results show that the attachment of Pseudomonas aeruginosa and Staphylococcus aureus can be robustly predicted by chemoinformatic descriptors, suggesting that the obtained models can predict the attachment response to polyacrylates to identify anti-attachment materials to synthesize and test in the future.

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Salmonella enterica serovar Enteritidis is a common foodborne pathogen that infects both humans and animals. The S. Enteritidis virulence regulation network remains largely incomplete, and knowledge regarding the specific virulence phenotype of small RNAs (sRNAs) is limited. Here, we investigated the role of a previously identified sRNA, Salmonella adhesive-associated sRNA (SaaS), in the virulence phenotype of S. Enteritidis by constructing mutant (ΔsaaS) and complemented (ΔsaaS/psaaS) strains. SaaS did not affect S. Enteritidis; it was activated in the simulated intestinal environment (SIE), regulating the expression of virulence target genes. We discovered that it directly binds ssaV mRNA. Caco-2 and RAW 264.7 cell assays revealed that SaaS promoted S. Enteritidis invasion and damage to epithelial cells while suppressing macrophage overgrowth and destruction. Furthermore, a BALB/c mouse model demonstrated that the deletion of SaaS significantly reduced mortality and attenuated the deterioration of pathophysiology, bacterial dissemination into systemic circulation, and systemic inflammation. Our findings indicate that SaaS is required for S. Enteritidis virulence and further highlight its biological role in bacterial pathogenesis. IMPORTANCE Salmonella is a zoonotic pathogen with high virulence worldwide, and sRNAs have recently been discovered to play important roles. We explored the biological characteristics of the sRNA SaaS and developed two cell infection models and a mouse infection model. SaaS is an SIE-responsive sRNA that regulates the expression of virulence-targeted genes. Additionally, it differentially mediates invasion and intracellular growth for survival and infection of the epithelium and macrophages. We further found that SaaS enhanced bacterial virulence by promoting lethality, colonization, and inflammatory response. These findings provide a better understanding of the critical role of sRNA in bacterial virulence.

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Flagella and their property would influence the initial attachment of bacteria onto plastics, yet their impacts have not been investigated. In present study, four types of E. coli with or without flagella as well as with normal or sticky flagella were utilized to investigate the effects of flagella and their property on the initial attachment behaviors of bacteria onto six types of plastics in freshwater systems. We found that E. coli with flagella exhibited better initial attachment performance onto all six types of plastics than strain without flagella. Flagella could help bacteria swim near to plastics, pierce the energy barrier, and subsequently attach onto plastics. With stronger adhesive force, sticky flagella could further facilitate bacterial attachment onto plastics. Moreover, flagella especially sticky flagella could help bacteria form more rigid attachment layer on plastics. Even with humic acid in suspensions or in river water, flagellar E. coli showed greater attachment onto plastics than E. coli without flagella. Humic acid might adsorb onto sticky flagella and thus decreased the attachment of bacteria with sticky flagella onto plastics. Obviously, flagella as well as their property would impact the initial attachment of bacteria onto plastics and the subsequent formation of plastisphere in freshwater.

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Different bacterial isolates attach to the cuticle of plant-parasitic nematodes, affecting their interactions with the host plant. Nematode populations differ in their genetic and cuticle structures, causing variable interactions with host plants and natural enemies. In the current study, attachment assays were carried out to compare the attachment of soil bacteria in general and the bacterial isolate of Rothia sp. in particular among geographically diverse populations of Pratylenchus neglectus. Biological and molecular assays were further conducted to examine the effect of Rothia attachment on nematode penetration into barley roots and to sequence the fatty acid- and retinol-binding gene (Pn-far-1). The results showed that nematode populations of P. neglectus differed in their bacterial attachment. Soil bacteria and Rothia sp. attached specifically to the cuticle of P. neglectus and did so differently among the nematode populations. Rothia attachment caused a reduction in the infectivity of three nematode populations in barley roots. The sequencing of the far-1 gene revealed genetic variability within and among P. neglectus populations. In conclusion, the interaction between P. neglectus and their bacterial attachers occurs in a population-specific manner, elucidating an essential aspect of using biological agents to manage plant-parasitic nematodes. Key Message: 1. Geographically diverse populations of the root lesion nematode Pratylenchus neglectus differed in the soil bacterial communities attached to their cuticles. 2. The bacterial isolate of Rothia sp. attached to the cuticle of P. neglectus and reduced its penetration into the host plant in a population-specific manner. 3. The fatty acid- and retinol-binding gene (far-1) varied within and among P. neglectus populations with their different bacterial attachment.

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ABSTRACT: Biofilm formation of Listeria monocytogenes on stainless steel, a widely used abiotic surface in the food processing industry, was investigated by focusing on the attachment tendency and behavior of L. monocytogenes 08-5578 on eight different stainless steel surfaces: glass bead blasted (rough and fine), deburred (Timesaver), drum deburred, pickled, pickled and drum polished, electrolytic polished, and cold rolled (untreated control). The aim was to see whether there are finishes with significantly lower bacterial attachment. Surface roughness data (measured via four roughness parameters), determined by interferometry, was also compared with the number of adhering cells to detect possible correlations. Cultivation of L. monocytogenes biofilms was carried out using a CDC biofilm reactor with 1% tryptic soy broth set at 20°C for 4, 8, and 24 h. In addition, a cultivation trial was run with continuous nutrient flow (1% tryptic soy broth, 6.2 mL/min) for 24 h. Eight-hour results showed a significant difference (P < 0.05) in biofilm cell counts in biofilms between the glass bead-blasted surfaces (3.23 and 3.26 log CFU/cm2 for the fine and rough, respectively) and deburred (Timesaver) surface (2.57 log CFU/cm2), between drum deburred and deburred (Timesaver) surface (3.41 versus 2.57 log CFU/cm2), and between drum deburred and pickled surface (3.41 versus 2.77 log CFU/cm2). Data gained after 4-h, 24-h, and 24-h plus an additional 24-h continuous flow cultivation showed no significant difference in attachment among surfaces. No correlation between roughness data and attachment was found after all four incubation times, suggesting that roughness values, at these ranges, are insufficient in determining the surfaces' affinity to bacteria. Overall, this study suggests that roughness values cannot be used to predict the degree of L. monocytogenes attachment to a specific stainless steel surface.

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Genomic integration of genes and pathway-sized DNA cassettes is often an indispensable way to construct robust and productive microbial cell factories. For some uncommon microbial hosts, such as Mycolicibacterium and Mycobacterium species, however, it is a challenge. Here, we present a multiplexed integrase-assisted site-specific recombination (miSSR) method to precisely and iteratively integrate genes/pathways with controllable copies in the chromosomes of Mycolicibacteria for the purpose of developing cell factories. First, a single-step multi-copy integration method was established in M. neoaurum by a combination application of mycobacteriophage L5 integrase and two-step allelic exchange strategy, the efficiencies of which were ∼100% for no more than three-copy integration events and decreased sharply to ∼20% for five-copy integration events. Second, the R4, Bxb1 and ΦC31 bacteriophage Att/Int systems were selected to extend the available integration toolbox for multiplexed gene integration events. Third, a reconstructed mycolicibacterial Xer recombinases (Xer-cise) system was employed to recycle the selection marker of gene recombination to facilitate the iterative gene manipulation. As a proof of concept, the biosynthetic pathway of ergothioneine (EGT) in Mycolicibacterium neoaurum ATCC 25795 was achieved by remodeling its metabolic pathway with a miSSR system. With six copies of the biosynthetic gene clusters (BGCs) of EGT and pentose phosphate isomerase (PRT), the titer of EGT in the resulting strain in a 30 mL shake flask within 5 days was enhanced to 66 mg/L, which was 3.77 times of that in the wild strain. The improvements indicated that the miSSR system was an effective, flexible, and convenient tool to engineer the genomes of Mycolicibacteria as well as other strains in the Mycobacteriaceae due to their proximate evolutionary relationships.

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Fresh cucumbers have been recognized as a vehicle in foodborne disease outbreaks since several multistate outbreaks of salmonellosis linked to fresh cucumbers occurred in the United States. Little is known about how microbial cell surface characteristics that are known to affect adhesion can influence bacterial cross-contamination and transfer. This study investigated the role of S. Newport cell surface components on bacterial attachment and transfer in cucumbers. Wild type Salmonella Newport and its transposon mutants were used to inoculate cucumbers. Attachment strength of S. Newport wild type to cucumber was not significantly different than that of mutants. Log10 percent transfer of mutant strains to edible flesh was not different from the wild type. Significantly less wild type Salmonella remained on the peel and transferred to the peeler than one mutant did, but not the other. Our results suggest that while curli and cellulose enhance Salmonella attachment to surface of cucumbers, there appear to be other mechanisms and factors that govern Salmonella transfer in cucumbers.

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Bacterial infections related to medical devices can cause severe problems, whose solution requires in-depth understanding of the interactions between bacteria and surfaces. This work investigates the influence of surface physicochemistry on bacterial attachment and detachment under flow through both empirical and simulation studies. We employed polydimethylsiloxane (PDMS) substrates having different degrees of crosslinking as the model material and the extended Derjaguin - Landau - Verwey - Overbeek model as the simulation method. Experimentally, the different PDMS materials led to similar numbers of attached bacteria, which can be rationalized by the identical energy barriers simulated between bacteria and the different materials. However, different numbers of residual bacteria after detachment were observed, which was suggested by simulation that the detachment process is determined by the interfacial physicochemistry rather than the mechanical property of a material. This finding is further supported by analyzing the bacteria detachment from PDMS substrates from which non-crosslinked polymer chains had been removed: similar numbers of residual bacteria were found on the extracted PDMS substrates. The knowledge gained in this work can facilitate the projection of bacterial colonization on a given surface.

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The initial step of biofilm formation is bacteria attachment to biotic or abiotic surfaces and other bacteria through intra or interspecies interactions. Adhesion can be influenced by physicochemical conditions of the environment, such as iron. There is no available mathematical model of bacterial attachment giving realistic initiation rather than random adhesion. We describe a simple stochastic attachment model, from the simplest case in two dimensions with one bacterial species attaching on a homogeneous flat surface to more complex situations, with either several bacterial species, inhomogeneous or non-flat surfaces, or in three dimensions. The model depends on attachment probabilities (on the surface, laterally, or vertically on bacteria). Effects of each of these parameters were analyzed. This mathematical model is then applied to experimental oral microcolonies of Porphyromonas gingivalis, Streptococcus gordonii, and Treponema denticola, either as mono-, two, or three species, under different iron concentrations. The model allows to characterize the adhesion of three bacterial species and explore the effect of iron on attachment. This model appears as a powerful tool for initial attachment analysis of bacterial species. It will enable further modeling of biofilm formation in later steps with biofilm initialization more relevant to real-life subgingival biofilms.

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The interaction between A-type interflavan bonds from cranberry proanthocyanidins (PAC) and surface virulence factors of extra-intestinal pathogenic Escherichia coli (ExPEC) was studied. Electrospun nanofibers (ESNF) were fabricated using PAC and polycaprolactone (PCL) solutions and their physical and chemical properties were characterized. The ability of PAC:PCL composite ESNF to interact with and entrap ExPEC strain 5011 (ExPEC-5011) was evaluated in vitro by plate culturing and when formulated as a biofilter and nanocoating. As a biofilter, the PAC:PCL ESNF exhibited a dose-dependent ability to entrap ExPEC-5011. Images from scanning electron and fluorescent microscopies revealed that ESNF sections with higher amounts of PAC led to higher bacterial entrapment. The effectiveness PAC:PCL ESNF to bind ExPEC when applied as a nanocoating was studied using ESNF-coated polyvinyl chloride intermittent catheter. Results indicate that ExPEC-5011 was entrapped well into the PAC:PCL ESNF coating on the catheter. Overall, our results suggest that incorporating the biomolecule PAC in ESNF is a potential means for applications requiring bacterial entrapment, such as biofunctionalization, biofiltration, and surface coating, among others.

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The polycyclic aromatic hydrocarbons (PAHs) that enter the aqueous phase usually coexist with fulvic acid (FA). Therefore, we initiated this investigation to explore the influences of FA on bacterial biofilm formation and its potential to biodegrade pyrene (PYR), using electron microscopic techniques and isobaric tags for relative and absolute quantification (iTRAQ). Our results revealed that FA stimulated biofilm formation and enhanced the biodegradation of PYR. First, FA favored the three-dimensional proliferation of bacteria, with an OD590/OD600 value of up to 14.78, and the extracellular surfaces covered by a layer of biomaterials. Distinctive intracellular morphologies of texture and organization were accompanied by reduced inter-bacterial distances of less than 0.31 µm. The biofilms formed displayed interactions between FA and surficial proteins, as noted by band shifts for the C-O and CO groups. Strikingly, FA triggered the upregulation of 130 proteins that were either operational in biofilm formation or in metabolic adjustments; with the changes supported by the increasing intensity of free amino acids and the newly generated N-O bonds. The results above revealed that the enhanced biodegradation was related to the up-regulation of the proteins functioned for ribosomal and carbon metabolism, and the ultra-structural changes in FA-induced biofilm system.

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Various strategies to accelerate the formation of biofilms on minerals have been studied, and one of them is the use of D-galactose as an inducer of EPS production in planktonic cells of biooxidant bacteria. With the aim to evaluate the influence on the attachment and the effect over the solubilization of a polymetallic mineral concentrate, the behavior of a microbial consortium formed by Acidithiobacillus thiooxidans DSM 14887T and Leptospirillum ferrooxidans DSM 2705T previously induced with D-galactose for the early formation of EPS was studied. These microorganisms were previously adapted to 0.15 and 0.25% of D-galactose, respectively; afterward, different proportions of both strains were put in contact with the particle surface of a concentrate of polymetallic mineral. Also, to evaluate the affinity of each bacterium to the mineral, attachment tests were carried out with one of these species acting as a pre-colonizer. The same consortia were used to evaluate the solubilization of the polymetallic mineral. The results obtained show that the induction by D-galactose increases the microbial attachment percentage to the mineral by at least 10% with respect to the control of non-adapted consortia. On the other hand, the tests carried out with pre-colonization show that the order of inoculation also affects the microbial attachment percentage. From the different proportions tested, it was determined that the use of a consortium with a proportion of 50% of each species previously adapted to D-galactose and inoculated simultaneously, present a microbial attachment percentage to the mineral greater than 95% and better solubilization of a polymetallic mineral, reaching values of 9.7 and 11.7mgL-1 h-1 of Fe3+ and SO4 2-, respectively. Therefore, the use of D-galactose in small concentrations as inducer of EPS in acidophilic cells and the selection of an adequate strategy of inoculation can be beneficial to improve biooxidation since it would allow this process to develop in a shorter time by achieving a greater number of attached cells in a shorter time accelerating the solubilization of a sulfide mineral. Graphical AbstractEPS production using D-galactose as inducer and its influence in the attachment of consortia formed by differents proportions of A. thiooxidans and L. ferrooxidans inoculated at the same time and when one of them acting as a pre-colonizer.