Promote Biofilm Formation Research Articles

A Novel Gene vp0610 Negatively Regulates Biofilm Formation in Vibrio parahaemolyticus.

Vibrio parahaemolyticus is an important foodborne pathogen and its biofilm formation ability facilitates its colonization and persistence in foods by protecting it from stresses including environmental variation and antibiotic exposure. Several important proteins are involved in biofilm formation; however, the identity and function of many remain unknown. In this study, we discovered a hypothetical protein, VP0610 that negatively regulates biofilm formation in Vibrio parahaemolyticus, and we found that the loss of vp0610 typically results in pleiotropic phenotypes that contribute toward promoting biofilm formation, including significantly increased insoluble exopolysaccharide production and swimming motility, decreased soluble exopolysaccharide production, and decreased bis-(3′-5′)-cyclic dimeric guanosine monophosphate production. Pull-down assays revealed that VP0610 can interact with 180 proteins, some of which (Hfq, VP0710, VP0793, and CyaA) participate in biofilm formation. Moreover, deleting vp0610 enhanced the expression of genes responsible for biofilm component (flaE), the sugar phosphotransferase system (PTS) EIIA component (vp0710 and vp0793), and a high-density regulator of quorum sensing (opaR), while reducing the expression of the bis-(3′-5′)-cyclic dimeric guanosine monophosphate degradation protein (CdgC), resulting in faster biofilm formation. Taken together, our results indicate that vp0610 is an integral member of the key biofilm regulatory network of V. parahaemolyticus that functions as a repressor of biofilm formation.

Mining for mouth metabolites.

Genome mining of Streptococcus mutans clinical samples led to the identification of a structurally unique polyketide/non-ribosomal peptide, mutanofactin-697, that promotes biofilm formation. This study highlights the unexplored functional potential of secondary metabolites and opens new avenues for inhibiting pathogenic processes.

The Topical Tranexamic Acid Have Potential Hazard of Promoting Biofilm Formation of Staphylococcus aureus in Microenvironment of the Prosthetic Joint

Background. Perioperative topical tranexamic acid as antifibrinolytic agent is often used for total joint replacement to reduce bleeding currently. Staphylococcus aureus was the most common isolates from perioperative infection of prosthetic joint. The influence of topical application with tranexamic acid on the incidence of acute prosthetic joint infection of Staphylococcus aureus has not been clarified. Methods. Mouse model of Staphylococcus aureus knee prosthesis infection was constructed. Tranexamic acid was intra-articular injected during the perioperative period. CFU counting from tissue and implant sample was evaluated 3 days and 7 days after inoculating of Staphylococcus aureus. Bacterial growth curve, biofilm formation, aggregation, and plasmin inhibition of Staphylococcus aureus were tested with tranexamic acid added to the synovial culture medium. Results. There were no significant differences of CFU counting from tissue and implant samples in knee prosthesis infection after a single local injection of tranexamic acid at the postoperative 3 or 7 days. The amount of bacterial colonization on the surface of implant increased after 3 days’ continuous local injection of tranexamic acid. Tranexamic acid has no effect on bacterial growth at the concentration (10 mg/ml) of clinical application, but it can inhibit bacterial aggregation and mildly inhibit biofilm formation. Plasmin can significantly inhibit biofilm formation which can be revised by adding tranexamic acid. Conclusion. Although continuous local injection of tranexamic acid can promote the biofilm formation of Staphylococcus aureus on the surface of articular implant, it has clinical safety for using one single local injection of tranexamic acid during the perioperative period.

Mutanofactin promotes adhesion and biofilm formation of cariogenic Streptococcus mutans.

Cariogenic Streptococcus mutans is known as a predominant etiological agent of dental caries due to its exceptional capacity to form biofilms. From strains of S. mutans isolated from dental plaque, we discovered, in the present study, a polyketide/nonribosomal peptide biosynthetic gene cluster, muf, which directly correlates with a strong biofilm-forming capability. We then identified the muf-associated bioactive product, mutanofactin-697, which contains a new molecular scaffold, along with its biosynthetic logic. Further mode-of-action studies revealed that mutanofactin-697 binds to S. mutans cells and also extracellular DNA, increases bacterial hydrophobicity, and promotes bacterial adhesion and subsequent biofilm formation. Our findings provided an example of a microbial secondary metabolite promoting biofilm formation via a physicochemical approach, highlighting the importance of secondary metabolism in mediating critical processes related to the development of dental caries.

New Bacterial Surface Display System Development and Application Based on Bacillus subtilis YuaB Biofilm Component as an Anchoring Motif.

Bacterial surface display system has been adopted in various biotechnological applications. In the case of Bacillus subtilis, most of the studies have been developed using spore based surface display system utilizing the inherent rigidity of spore against heat, alkali, and shear stress. But, spore harvest, purification and separation need additional cost and labor. To eliminate this procedure and to use the gram-positive nature of B. subtilis, YuaB, which is one of the major B. subtilis biofilm components and locates in the cell wall, based cell surface display system, is developed. P43 promoter driven overexpression of YuaB-His6 tag does not hamper bacterial cell growth and promoted biofilm formation of recombinant strain. Flow cytometry of recombinant strain and its protoplast using FITC-Anti His6 antibody, verified that YuaB locate in plasma membrane and protrude to the outside of cell wall, which means YuaB can be used as very efficient anchoring motif. Using surface expressed YuaB-His6 tag, removal of divalent metal ion, Cu2+ and Ni2+, was tried to test its possibility for the environmental application of developed system.

Starvation Survival and Biofilm Formation under Subminimum Inhibitory Concentration of QAMs.

Quaternary ammonium methacrylates (QAMs) are useful antimicrobial compounds against oral bacteria. Here, we investigated the effects of two QAMs, dimethylaminododecyl methacrylate (DMADDM) and dimethylaminohexadecyl methacrylate (DMAHDM), on biofilm formation, survival and development of tolerance by biofilm, and survival and development of tolerance against QAMs after prolonged starvation. Enterococcus faecalis (E. faecalis), Streptococcus gordonii (S. gordonii), Lactobacillus acidophilus (L. acidophilus), and Actinomyces naeslundii (A. naeslundii) were used. Minimum inhibitory concentration (MIC) of QAMs against multispecies biofilm was determined. Biofilm formed under sub-MIC was observed by crystal violet staining and confocal laser scanning microscopy (CLSM). Metabolic activity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactic acid production measurement. Development of tolerance was determined by MIC values before and after exposure to QAMs or after prolonged starvation. It was found that E. faecalis and S. gordonii could survive and form biofilm under sub-MIC of QAMs. Lactic acid production from biofilms formed under sub-MIC was significantly higher than control specimens (p < 0.05). The exposure to sub-MIC of QAMs promoted biofilm formation, and prolonged starvation or prolonged contact with sub-MIC helped bacteria develop tolerance against killing by QAMs.

Outer Membrane Vesicles Released From Aeromonas Strains Are Involved in the Biofilm Formation.

Aeromonas spp. are Gram-negative rod-shaped bacteria ubiquitously distributed in diverse water sources. Several Aeromonas spp. are known as human and fish pathogens. Recently, attention has been focused on the relationship between bacterial biofilm formation and pathogenicity or drug resistance. However, there have been few reports on biofilm formation by Aeromonas. This study is the first to examine the in vitro formation and components of the biofilm of several Aeromonas clinical and environmental strains. A biofilm formation assay using 1% crystal violet on a polystyrene plate revealed that most Aeromonas strains used in this study formed biofilms but one strain did not. Analysis of the basic components contained in the biofilms formed by Aeromonas strains confirmed that they contained polysaccharides containing GlcNAc, extracellular nucleic acids, and proteins, as previously reported for the biofilms of other bacterial species. Among these components, we focused on several proteins fractionated by SDS-PAGE and determined their amino acid sequences. The results showed that some proteins existing in the Aeromonas biofilms have amino acid sequences homologous to functional proteins present in the outer membrane of Gram-negative bacteria. This result suggests that outer membrane components may affect the biofilm formation of Aeromonas strains. It is known that Gram-negative bacteria often release extracellular membrane vesicles from the outer membrane, so we think that the outer membrane-derived proteins found in the Aeromonas biofilms may be derived from such membrane vesicles. To examine this idea, we next investigated the ability of Aeromonas strains to form outer membrane vesicles (OMVs). Electron microscopic analysis revealed that most Aeromonas strains released OMVs outside the cells. Finally, we purified OMVs from several Aeromonas strains and examined their effect on the biofilm formation. We found that the addition of OMVs dose-dependently promoted biofilm formation, except for one strain that did not form biofilms. These results suggest that the OMVs released from the bacterial cells are closely related to the biofilm formation of Aeromonas strains.

Pathogenic bacterial toxins and virulence influences in cultivable fish

Bacterial virulence factors play a major role in infection, which provides devastating effect on the aquaculture industry. Bacterial toxins are the major virulence factors that influence the roles of host cells and take control of living organisms’ vital processes to facilitate microbial infection. Various other bacterial virulent components such as adhesins, capsular polysaccharides, lipopolysaccharides, bacterial flagellum, pili and microbial siderophore provide a diverse effect in the microbial pathogenesis and express a variety of pathogenic molecules including aebABCEG, HutA, cpsD and pPHDD1. Bacterial protein secretion system secretes enzymes and toxins from bacterial cytoplasm to the host membrane targeting the immune cells and promoting biofilm formation. However, as they enter, the host body expresses various immune molecules that interact to utilize the pathogen recognition receptors. Toxins are powerful molecules formed by a broad range of bacterial pathogens attacking host cells and playing key roles in the dialogue between host and pathogen. They are important virulence factors that are often enough to decide the outcome of the infection. Hence, a detailed view on the virulence molecules majorly exotoxins from various bacterial pathogenic strains that are necessary to understand the pathogenesis. This review focuses on bacterial toxins and various other virulence factors, which influence the pathogenesis in fish.

Foliar pathogen-induced assemblage of beneficial rhizosphere consortia increases plant defense against Setosphaeria turcica.

Background: Foliar pathogen infection can induce the enrichment of beneficial microbial consortia in plant rhizosphere, but the mechanism for enhanced plant resistance is unclear. Methods: We investigated the effects of foliar pathogen infection on bacterial communities in maize rhizosphere using high throughput sequencing. Results: Maize plants grown in non-sterilized soils displayed stronger defense against the foliar pathogen Setosphaeria turcica than those in sterilized soils. Foliar pathogen infection further triggered the shift in the structure and composition of rhizosphere bacterial communities. The pathogen-infected plants specially promoted rhizosphere colonization of several bacterial taxa. The Pseudomonas genus increased in the rhizosphere after pathogen infection. Other bacterial genera such as Chitinophaga and Flavobacterium were also greatly enriched in the rhizosphere of pathogen-infected plants. Furthermore, the enriched bacterial species were isolated and were shown to interact synergistically to promote biofilm formation. Although both the Chitinophaga and Flavobacterium species did not induce plant defense, the Pseudomonas species markedly increased the resistance of plants against S. turcica. Furthermore, the consortium consisting of the Pseudomonas, Chitinophaga and Flavobacterium species (CONpcf) conferred long-acting disease resistance of maize plants as compared to the individual Pseudomonas species. Furthermore, the inoculation with the CONpcf significantly induced a marked increase in the levels of DIMBOA in maize leaves, indicating that the consortium-induced increases of DIMBOA levels partially contributed to enhancing disease resistance of plants. Conclusions: Foliar infection of maize plants by S. turcica specifically recruited a group of beneficial rhizosphere bacteria, which conferred enhanced plant defense against pathogen infection. This study provided important evidence that above-ground pathogen infection participated in the mediation of below-ground microbiome for regulating plant defense systems.

Evolutionary trade-offs between growth and survival: The delicate balance between reproductive success and longevity in bacteria.

All living cells strive to allocate cellular resources in a way that promotes maximal evolutionary fitness. While there are many competing demands for resources the main decision making process centres on whether to proceed with growth and reproduction or to "hunker down" and invest in protection and survival (or to strike an optimal balance between these two processes). The transcriptional programme active at any given time largely determines which of these competing processes is dominant. At the top of the regulatory hierarchy are the sigma factors that commandeer the transcriptional machinery and determine which set of promoters are active at any given time. The regulatory inputs controlling their activity are therefore often highly complex, with multiple layers of regulation, allowing relevant environmental information to produce the most beneficial response. The tension between growth and survival is also evident in the developmental programme necessary to promote biofilm formation, which is typically associated with low growth rates and enhanced long-term survival. Nucleotide second messengers and energy pools (ATP/ADP levels) play critical roles in determining the fate of individual cells. Regulatory small RNAs frequently play important roles in the decision making processes too. In this review we discuss the trade-off that exists between reproduction and persistence in bacteria and discuss some of the recent advances in this fascinating field.

Onychomycosis as a Chronic Superficial Fungal Infection: "Why is it so Hard to Clear?"

Onychomycosis is a fungal infection of the nail unit. Compared to other superficial dermatophytoses of the skin and hair, onychomycosis is associated with a chronic course and frequent recurrence. Originally, structural characteristics of the nail, such as poor vascular supply and thick layers of hard keratin, were thought to be limiting factors for efficient drug delivery, resulting in prolonged treatment. However, recent research reveals the following crucial mechanisms contributing to the chronicity of nail dermatophytosis: innate characteristics of Trichophyton rubrum, formation of dormant spores called arthroconidia, physical and immunologic characteristics of the nail apparatus, and alteration of the defense system of the host. T. rubrum, the most common causative fungal pathogen for onychomycosis, acquires selective immune tolerance toward the host's defense system. Transformation into dormant phases, such as arthroconidia, creates greater resistance to antifungal medication and nonnutritive environments. The anatomical and biological structures of the nail allow immune evasion, increase infection susceptibility, and promote biofilm formation. Key regulators of the host's innate and adaptive immune systems are downregulated as infection persists, and age-associated immunomodulation aggravates the process.

Indole-3-acetic acid synthesized through the indole-3-pyruvate pathway promotes Candida tropicalis biofilm formation.

We previously found that the elevated abundance of the fungus Candida tropicalis is positively correlated with the bacteria Escherichia coli and Serratia marcescens in Crohn's disease patients and the three pathogens, when co-cultured, form a robust mixed-species biofilm. The finding suggests that these three pathogens communicate and promote biofilm formation, possibly through secretion of small signaling molecules. To identify candidate signaling molecules, we carried out a metabolomic analysis of the single-species and triple-species cultures of the three pathogens. This analysis identified 15 metabolites that were highly increased in the triple-species culture. One highly induced metabolite was indole-3-acetic acid (IAA), which has been shown to induce filamentation of certain fungi. We thus tested the effect of IAA on biofilm formation of C. tropicalis and demonstrated that IAA promotes biofilm formation of C. tropicalis. Then, we carried out isotope tracing experiments using 13C-labeled-tryptophan as a precursor to uncover the biosynthesis pathway of IAA in C. tropicalis. The results indicated that C. tropicalis synthesizes IAA through the indole-3-pyruvate pathway. Further studies using inhibitors of the indole-3-pyruvate pathway are warranted to decipher the mechanisms by which IAA influences biofilm formation.

Effect of water aging on the anti-biofilm properties of glass ionomer cement containing fluoro-zinc-silicate fillers

A previous study has reported that a novel fluoro-zinc-silicate glass ionomer cement (Caredyne Restore) showed superior anti-biofilm effects by interfering with bacterial adhesion. However, the active ions may degrade with time. This study aimed to assess the valid anti-biofilm effects of Caredyne Restore after being aged by water immersion for 3 weeks. Streptococcus mutans biofilm was allowed to grow on the surface before and after water aging for 24 h using a modified Robbins device flow-cell system. The results showed water aging promoted biofilm formation. Insufficient amount of fluoride and zinc ions were released from Caredyne Restore after water aging under neutral pH condition. An acidic pH is needed to exert effective anti-biofilm properties. As the release of active ions from Caredyne Restore will gradually decrease after the restoration, the restoration may not prevent biofilm formation after 3 weeks while neutral pH is maintained by the buffering capacity of saliva.

Virtual Screening for Novel SarA Inhibitors to Prevent Biofilm Formation of Staphylococcus aureus in Prosthetic Joint Infections.

Staphylococcus aureus is one of the predominant causes of periprosthetic joint infections (PJIs). Bacterial adhesion and biofilm formation are important factors in the pathogenesis of PJIs. S. aureus biofilm formation is regulated by several factors, including S. aureus regulator A (SarA). Previous studies have found that SarA mutants have limited ability to develop biofilms. In this study, we identified a SarA-targeting antibiofilm compound, ZINC00990144, and evaluated its efficacy and toxicity. According to static biofilm assay, the antibiofilm ability of the compound was concentration dependent. ZINC00990144 reduced biofilm in multiple strains by 40–86% at a concentration of 11.5 μM. Additionally, ZINC00990144 inhibited biofilm formation on different orthopedic implant materials including Titanium and UHMWPE disc. Furthermore, quantitative polymerase chain reaction results demonstrated that ZINC00990144 upregulated the expression of S. aureus exoproteases to inhibit the formation of biofilms. Moreover, ZINC00990144 prevented biofilm formation when exposed to sub-inhibitory doses of vancomycin, which is known to promote biofilm formation. CCK-8 results demonstrated ZINC00990144 has no significant effect on cell viability at concentration of 11.5 μM or below. Finally, we verified the antibiofilm function of the compound in vivo using implant infection mice model with/without exposure to sub-inhibitory vancomycin. In conclusion, ZINC00990144 acts by modulating between biofilm and planktonic state of S. aureus instead of being bactericidal. Therefore, it has the potential to be used in combination with other antibiotics to prevent PJIs.

Elevated c-di-GMP levels promote biofilm formation and biodesulfurization capacity of Rhodococcus erythropolis.

Bacterial biofilms provide high cell density and a superior adaptation and protection from stress conditions compared to planktonic cultures, making them a very promising approach for bioremediation. Several Rhodococcus strains can desulfurize dibenzothiophene (DBT), a major sulphur pollutant in fuels, reducing air pollution from fuel combustion. Despite multiple efforts to increase Rhodococcus biodesulfurization activity, there is still an urgent need to develop better biocatalysts. Here, we implemented a new approach that consisted in promoting Rhodococcus erythropolis biofilm formation through the heterologous expression of a diguanylate cyclase that led to the synthesis of the biofilm trigger molecule cyclic di-GMP (c-di-GMP). R. erythropolis biofilm cells displayed a significantly increased DBT desulfurization activity when compared to their planktonic counterparts. The improved biocatalyst formed a biofilm both under batch and continuous flow conditions which turns it into a promising candidate for the development of an efficient bioreactor for the removal of sulphur heterocycles present in fossil fuels.

Phenotypic and proteomic approaches of the response to iron-limited condition in Staphylococcus lugdunensis

BackgroundStaphylococcus lugdunensis is a coagulase-negative Staphylococcus part of the commensal skin flora but emerge as an important opportunistic pathogen. Because iron limitation is a crucial stress during infectious process, we performed phenotypic study and compared proteomic profiles of this species incubated in absence and in presence of the iron chelator 2,2′-dipyridyl (DIP).ResultsNo modification of cell morphology nor cell wall thickness were observed in presence of DIP. However iron-limitation condition promoted biofilm formation and reduced the ability to cope with oxidative stress (1 mM H2O2). In addition, S. lugdunensis N920143 cultured with DIP was significantly less virulent in the larvae of Galleria mellonella model of infection than that grown under standard conditions. We verified that these phenotypes were due to an iron limitation by complementation experiments with FeSO4. By mass spectrometry after trypsin digestion, we characterized the first iron-limitation stress proteome in S. lugdunensis. Among 1426 proteins identified, 349 polypeptides were differentially expressed. 222 were more and 127 less abundant in S. lugdunensis incubated in iron-limitation condition, and by RT-qPCR, some of the corresponding genes have been shown to be transcriptionally regulated. Our data revealed that proteins involved in iron metabolism and carriers were over-expressed, as well as several ABC transporters and polypeptides linked to cell wall metabolism. Conversely, enzymes playing a role in the oxidative stress response (especially catalase) were repressed.ConclusionsThis phenotypic and global proteomic study allowed characterization of the response of S. lugdunensis to iron-limitation. We showed that iron-limitation promoted biofilm formation, but decrease the oxidative stress resistance that may, at least in part, explained the reduced virulence of S. lugdunensis observed under low iron condition.

Biosynthetic FeS/BC hybrid particles enhanced the electroactive bacteria enrichment in microbial electrochemical systems

Modifying the surface of an anode can improve electroactive bacteria (EAB) enrichment, thereby enhancing the performance of the associated microbial electrochemical systems (MESs). In this study, biosynthetic FeS nanoparticles were used to modify the anode in MESs. The experimental results demonstrated that the stable maximum voltage of the FeS composited biochar (FeS/BC)-modified anode reached 0.72 V, which is 20% higher than that of the control. The maximum power density with the FeS/BC anode was 793 mW/m2, which is 46.31% higher than that obtained with the control (542 mW/m2). According to cyclic voltammetry (CV) analysis, FeS/BC facilitates the direct electron transfer between bacteria and the electrode. The biomass protein concentration of the FeS/BC anode was 841.75 μg/cm2, which is almost 1.5 times higher than that of the carbon cloth anode (344.25 μg/cm2); hence, FeS/BC modification can promote biofilm formation. The composition of Geobacter species on the FeS/BC anode (75.16%) was much higher than that on the carbon cloth anode (4.81%). All the results demonstrated that the use of the biosynthetic FeS/BC anode is an environmentally friendly and efficient strategy for enhancing the electroactive biofilm formation and EAB enrichment in MESs.

Degradation of subµ-sized bioplastics by clinically important bacteria under sediment and seawater conditions: Impact on the bacteria responses

In this study, we investigated the interaction of submicron-sized bioplastics with environmentally and clinically important bacteria under seawater and sediment conditions. To examine the relationship between submicron-sized bioplastics and bacteria in seawater and sediment, we focused on the bacterial activation and their biochemical key events toward the protein, carbohydrate, lipid, and antioxidant response. In addition, culture-dependent biofilm formation on submicron-sized bioplastics and their characterization was performed. The results indicated that selected bacteria increased their viability both in seawater and sediment with the submicron-sized bioplastics in that the bioplastics decreased their mass at the level of 10–23%. However, the activation level and mechanism affected the polymer type, bacteria, and environmental media, and submicron-sized bioplastics promoted biofilm formation with enhancing basophilic characteristics of biofilms.

Improving the freeze-drying survival rate of Lactobacillus plantarum LIP-1 by increasing biofilm formation based on adjusting the composition of buffer salts in medium

Lactic acid bacteria can improve their resistance to adverse environments through the formation of biofilm. This study found that adding different buffer salts in culture medium had a great impact on the freeze-drying survival rate of the Lactobacillus plantarum LIP-1, which could be linked to biofilm formation. Transcriptome data showed that potassium ions in buffer salt increased the expression of the luxS gene in the LuxS/autoinducer-2 (AI-2) quorum sensing system and increase synthesis of the quorum sensing signal AI-2. The AI-2 signal molecules up-regulated the cysE gene, which helps to promote biofilm formation. By adding a biofilm inhibitor, d-galactose, and performing a real-time quantitative polymerase chain reaction experiment, we found that d-galactose could down-regulated the luxS and cysE genes, reduced biofilm formation, and decreased the freeze-drying survival rate. The results of this study showed that promoting biofilm formation using appropriate buffer salts may lead to better freeze-drying survival rates.

Metabolites Produced by Kaistia sp. 32K Promote Biofilm Formation in Coculture with Methylobacterium sp. ME121.

Previously, we reported that the coculture of motile Methylobacterium sp. ME121 and non-motile Kaistia sp. 32K, isolated from the same soil sample, displayed accelerated motility of strain ME121 due to an extracellular polysaccharide (EPS) produced by strain 32K. Since EPS is a major component of biofilms, we aimed to investigate the biofilm formation in cocultures of the two strains. The extent of biofilm formation was measured by a microtiter dish assay with the dye crystal violet. A significant increase in the amount of biofilm was observed in the coculture of the two strains, as compared to that of the monocultures, which could be due to a metabolite produced by strain 32K. However, in the coculture with strain 32K, using Escherichia coli or Pseudomonas aeruginosa, there was no difference in the amount of biofilm formation as compared with the monoculture. Elevated biofilm formation was also observed in the coculture of strain ME121 with Kaistia adipata, which was isolated from a different soil sample. Methylobacterium radiotolerans, isolated from another soil sample, showed a significant increase in biofilm formation when cocultured with K. adipata, but not with strain 32K. We also found that the culture supernatants of strains 32K and K. adipata accelerated the motility of strains ME121 and M. radiotolerans, wherein culture supernatant of K. adipata significantly increased the motility of M. radiotolerans, as compared to that by the culture supernatant of strain 32K. These results indicated that there was a positive relationship between accelerated motility and increased biofilm formation in Methylobacterium spp. This is the first study to report that the metabolites from Kaistia spp. could specifically modulate the biofilm-forming ability of Methylobacterium spp. Methylobacterium spp. biofilms are capable of inhibiting the biofilm formation of mycobacteria, which are opportunistic pathogens that cause problems in infectious diseases. Thus, the metabolites from the culture supernatant of Kaistia spp. have the potential to contribute to the environment in which increased biofilm production of Methylobacterium is desired.