The clinical impact of bacterial biofilms

Bacteria survive in nature by forming biofilms on surfaces and probably most, if not all, bacteria （and fungi） are capable of forming biofilms. A biofilm is a structured consortium of bacteria embedded in a self-produced polymer matrix consisting of polysaccharide, protein and extracellular DNA. Bacterial biofilms are resistant to antibiotics, disinfectant chemicals and to phagocytosis and other components of the innate and adaptive inflammatory defense system of the body. It is known, for example, that persistence of staphylococcal infections related to foreign bodies is due to biofilm formation. Likewise, chronic Pseudomonas aeruginosa lung infections in cystic fibrosis patients are caused by biofilm growing mucoid strains. Gradients of nutrients and oxygen exist from the top to the bottom of biofilms and the bacterial cells located in nutrient poor areas have decreased metabolic activity and increased doubling times. These more or less dormant cells are therefore responsible for some of the tolerance to antibiotics. Biofilm growth is associated with an increased level of mutations. Bacteria in biofilms communicate by means of molecules, which activates certain genes responsible for production of virulence factors and, to some extent, biofilm structure. This phenomenon is called quorum sensing and depends upon the concentration of the quorum sensing molecules in a certain niche, which depends on the number of the bacteria. Biofilms can be prevented by antibiotic prophylaxis or early aggressive antibiotic therapy and they can be treated by chronic suppressive antibiotic therapy. Promising strategies may include the use of compounds which can dissolve the biofilm matrix and quorum sensing inhibitors, which increases biofilm susceptibility to antibiotics and phagocytosis.

Strategies for combating bacterial biofilm infections

Formation of biofilm is a survival strategy for bacteria and fungi to adapt to their living environment, especially in the hostile environment. Under the protection of biofilm, microbial cells in biofilm become tolerant and resistant to antibiotics and the immune responses, which increases the difficulties for the clinical treatment of biofilm infections. Clinical and laboratory investigations demonstrated a perspicuous correlation between biofilm infection and medical foreign bodies or indwelling devices. Clinical observations and experimental studies indicated clearly that antibiotic treatment alone is in most cases insufficient to eradicate biofilm infections. Therefore, to effectively treat biofilm infections with currently available antibiotics and evaluate the outcomes become important and urgent for clinicians. The review summarizes the latest progress in treatment of clinical biofilm infections and scientific investigations, discusses the diagnosis and treatment of different biofilm infections and introduces the promising laboratory progress, which may contribute to prevention or cure of biofilm infections. We conclude that, an efficient treatment of biofilm infections needs a well-established multidisciplinary collaboration, which includes removal of the infected foreign bodies, selection of biofilm-active, sensitive and well-penetrating antibiotics, systemic or topical antibiotic administration in high dosage and combinations, and administration of anti-quorum sensing or biofilm dispersal agents.

Phylogenetic diversity of bacterial biofilms covering the settlement substrates of nona-porous abalones (Haliotis diversicolor supertexta)

The settlement substrates of nona-porous abalones （Haliotis diversicolor supertexta） are covered with biofilms in which several types of microorganisms coexist and interact. These microorganisms are usually important causes of juvenile abalone disease as well as organisms useful in promoting abalones’ adhesion. The bacterial community structure of the biofilms remains unclear. The aim of this research was to determine the genetic diversity and phylogenetic affiliation of the biofilm bacteria. Total DNA of bacteria in biofilms was extracted, and 16S rRNA gene clone library was constructed using the primers specific for the domain bacteria. Subsequently, 30 randomly selected positive clones were screened by PCR-restriction fragment length polymorphism （PCR-RFLP） analysis, and resulted in 15 different RFLP patterns. Sequences analysis of representatives from each unique RFLP type revealed high genetic diversity in the bacterial populations. These sequences fell into nine major lineages of the bacterial domains： α-, β-, γ-and δ-subdivisions of the Proteobacteria; Planctomycete, Actinobacteria, Firmicutes, V errucomicrobium spp., and CytophagaFlexibacter-Bacteroides spp. Phylogenetic analysis indicated that the dominant phylotypes were most closely related to environmental and clinical Burkholderia cepacia of the β-Proteobacteria, and Roseobacteria of the α-Proteobacteria.

Bacterial Biofilm Formation on Resorbing Magnesium Implants

Background: Implant-associated infections are a result of bacterial adhesion to an implant surface and subsequent biofilm formation at the implantation site. This study compares different magnesium materials based on their ability to resist bacterial adhesion as well as further biofilm formation. Material and Methods: The surfaces of four magnesium-based materials (Mg2Ag, Mg10Gd, WE43 and 99.99% pure Mg) were characterized using atomic force microscope. In addition, the samples were tested for their ability to resist biofilm formation. Planktonic bacteria of either S. epidermidis or E. faecalis were allowed to adhere to the magnesium surfaces for two hour followed by rinsing and, for S. epidermidis, further incubation of 24, 72 and 168 h was carried out. Results: E. faecalis had a significantly stronger adhesion to all magnesium surfaces compared to S. epidermidis (p = 0.001). Biofilm growth of S. epidermidis was different on various magnesium materials: the amount of bacteria increased up to 72 h but interestingly a significant decrease was seen at 168 h on Mg2Ag and WE43 surfaces. For pure Mg and Mg10Gd the biofilm formation reached plateau at 72 h. Surface characteristics of resorbable magnesium materials were changing over time, and the surface was generally less rough at 168 h compared to earlier time points. No correlation was found between the surface topology and the amount of adherent bacteria. Conclusion: In early stages of biofilm adhesion, no differences between magnesium materials were observed. However, after 72 h Mg2Ag and WE43 had the best ability to suppress S. epidermidis’ biofilm formation. Also, bacterial adhesion to magnesium materials was not dependent on samples’ surface topology.

A polymyxin B loaded hypoxia-responsive liposome with improved biosafety for efficient eradication of bacterial biofilms

Biofilm-associated bacterial infection brings serious threats to global public health owing to serious antibiotic resistance.It is urgently needed to develop innovative strategies to combat biofilm-associated bacterial infections.Polymyxins stand out as the last line of defense against Gram-negative bacteria.However,serious nephrotoxicity of polymyxins severely limits their clinical utility.Herein,a hypoxia-responsive liposome is designed as the nanocarrier of polymyxin B(PMB)to combat biofilms developed by Gram-negative bacteria.A metronidazole modified lipid(hypoxia-responsive lipid(HRLipid))is synthesized to fabricate hypoxia-responsive liposomes(HRLip).PMB loaded hypoxia-responsive liposomes(HRL-PMB)is then prepared to mitigate the nephrotoxicity of PMB while preserving its excellent bactericidal activity.HRL-PMB shows very low hemolysis and cytotoxicity due to liposomal encapsulation of PMB.PMB can be readily released from HRL-PMB in response to hypoxic biofilm microenvironment,exerting its bactericidal activity to realize biofilm eradication.The excellent in vivo antibiofilm ability of HRL-PMB is confirmed by a Pseudomonas aeruginosa infected zebrafish model and a P.aeruginosa pneumonia infection model.Meanwhile,HRL-PMB can greatly reduce the nephrotoxicity of PMB after intravenous injection.The hypoxia-sensitive liposomes held great promise to improve the biosafety of highly toxic antibiotics while preserving their intrinsic bactericidal ability,which may provide an innovative strategy for combating biofilm-associated infections.

Comparison of MIC with MBEC Assay for in Vitro Antimicrobial Susceptibility Testing in Biofilm Forming Clinical Bacterial Isolates

Context: MIC results can be misleading for treatment of biofilm associated. The Minimum Biofilm Eradication Concentration (MBEC) measures the determination to be made for a biofilm susceptibility to antibiotics. Aims: Assessment of biofilm production and comparison of the MIC and MBEC assays evaluate differences in the antibiotic sensitivity patterns of different clinical bacterial isolates from patients implanted with medical devices. Settings and Design: Random sampling with experimental study at tertiary care institute. Methods and Material: The study was carried out during January 2014 to March 2014 on 50 positive bacteriological cultures of medical devices which were inserted in hospitalized patients. Biofilm forming strains were identified by tissue culture plate method & tube method. Biofilm-producing and non-biofilm forming reference strains were used as controls. Assay has been developed for the use with flat bottom, 96-well microtiter plates. Sterile autoclaved PCR tubes were used as pegs which provided surface for the biofilm formation. Amikacin, ciprofloxacin, trimethoprim-sulfamethoxazole, vancomycin, cefoperazone/ sulbactam, gentamycin were tested for MIC and MBEC assay. Statistical Analysis Used: Results will be discussed in the form of percentages. Results: Colonization by Klebsiella pneumoniae, Acinetobacter baumanni and Pseudomonas aeruginosa was prevalent bacterial isolates in medical devices. MBEC was higher for all the antibiotics as compared to MIC except amikacin MBEC for Pseudomonas was the same as MIC. Conclusions: Device associated bacterial biofilms are the major source of infections in patients of critical care setup. MIC misleads physician for organism’s drug susceptibility testing, which results in therapeutic failure. MBEC can guide regarding choice and proper dosing of antibiotics to be given. That’s why major studies for similar testing should be done with clinical evaluation.

Biofilm formation under high temperature causes the commensal bacteria Bacillus cereus WPySW2 to shift from friend to foe in Neoporphyra haitanensis in vitro model

Although biofilm formation may promote growth,biofilms are not always beneficial to their hosts.The biofilm formation characteristics of Bacillus cereus WPySW2 and its changes at different temperatures were studied.Results show that B.cereus WPySW2 promoted the growth of Neoporphyra haitanensis(an economically cultivated seaweed)at 20℃ but accelerated algal rot at 28℃.Thicker B.cereus WPySW2 biofilms covered the surface of N.haitanensis thalli at 28℃,which hindered material exchange between the algae and surrounding environment,inhibited algal photosynthesis and respiration,and accelerated algal decay.Compared with planktonic bacteria,mature biofilm cells had lower energy consumption and metabolic levels.The biofilm metabolic characteristics of B.cereus WPySW2 changed significantly with temperature.High temperature accelerated biofilm maturation,which made it thicker and more stable,allowing the bacteria to easily adapt to environmental changes and obtain greater benefits from their host.High temperature did not affect the production or increased the abundance of toxic metabolites,indicating that the negative effects of B.cereus WPySW2 on algae were not caused by toxins.This study shows that increased temperature can transform a harmless bacterium into a detrimental one,demonstrating that temperature may change the ecological function of phycospheric bacteria by affecting their morphology and metabolism.

Effects of carbon sources and temperature on the formation and structural characteristics of food-related Staphylococcus epidermidis biofilms

Biofilms are a constant concern in the food industry;understanding the effect of environmental conditions on biofilm formation is essential to develop effective control strategies.Therefore,this study was conducted to investigate biofilms formation by Staphylococcus epidermidis under various conditions.Biofilms were cultured in nutrient broth containing different carbon source concentrations(0–10 mg/mL)on polystyrene surfaces for 32 h of incubation at 37℃or 55℃,with quantification and enumeration at 8,16,24 and 32 h.S.epidermidis developed biofilms under all tested conditions;achieved the highest yield of biofilm biomass at 2.5 mg/mL for all carbon sources at 37℃.The highest efficiency of extracellular polymeric substance(EPS)molecule production occurred under glucose availability in the growth environment,with a higher yield of biomass and a significantly smaller number of metabolically active cells than under other tested conditions.A condensed ball-shaped structure was observed under the lactose condition.Meanwhile,biofilms in the presence of maltose showed mainly opaque thick rich colonies,while a compact multilayered-shaped structure was exhibited under both glucose and sucrose conditions.These results contribute to a better understanding of the biofilm formation by S.epidermidis in order to reduce contamination and recontamination in the food industry.

Novel Cathelicidins with Potent Antimicrobial,Biofilm Inhibitory,and Anti-inflammatory Activities from the Frog Fejervarya multistriata

Antimicrobial peptides（AMPs）,a class of gene-encoded peptides,are the first line of immune system to defense microbial invasions in multicellular organisms.Cathelicidins are an important family of AMPs that have been identified exclusively in vertebrates.However,up to now,cathelicidins from amphibians are poorly understood.In the present study,we reported the identification and characterization of two novel cathelicidins（FM-CATH1 and FMCATH2） from the frog Fejervarya multistriata.The c DNA sequences encoding FM-CATHs were successfully cloned from the constructed lung c DNA library of F.multistriata.Both of the c DNA sequences encoding FM-CATHs are 447 bp in length,and the deduced mature peptides of FM-CATHs are composed of 34 residues.Structural analysis indicated that FM-CATH1 and FM-CATH2 mainly assume amphipathic alpha-helical conformations.Antimicrobial and bacterial killing kinetic analysis indicated that both FM-CATH1 and FM-CATH2 possess potent,broad-spectrum and rapid antimicrobial potency.And cytoplasmic membrane permeabilization analysis indicated that FM-CATH1 and FMCATH2 kill bacteria by inducing the permeabilization of bacterial membrane.Besides direct antimicrobial activities,FM-CATHs also exhibited significant inhibitory effect on the formation of bacterial biofilms at low concentrations below 1×MIC.Furthermore,FM-CATH1 and FM-CATH2 exhibited potent anti-inflammatory activities by inhibiting LPS-induced transcription and production of pro-inflammatory cytokines TNF-α,IL-1β,and IL-6 in mouse peritoneal macrophages.Meanwhile,FM-CATHs showed relatively low cytotoxic activity against mammalian normal and tumor cell lines,and low hemolytic activity against human erythrocytes.In summary,the identification of FM-CATHs provides novel clues for our understanding of the roles of cathelicidins in amphibian immune systems.The potent antimicrobial,biofilm inhibitory,anti-inflammatory activities,and low cytotoxicity of FM-CATHs imply their great potential in novel antibiotics development.

Inhibition of Bacterial Adherence on the Surface of Biliary Stent Materials Modified With Chitosan

Bacterial infection plays an important role in the initiation of biliary sludge formation. Bacterial adherence and biofilm formation on the surface of a material have been considered as one of the main factors of stent re-occlusion in clinic. This work reported preventing bacterial adherence and bacterial biofilm formation on the surface of biliary stent material using chitosan film. The chitosan film was deposited on 316 L stainless steel （SS） plate by electrophoresis method and was characterized by X-ray diffraction （XRD）, Fourier Transform infrared spectroscopy （FTIR） and atomic force microscopy （AFM）. The ability of inhibiting bacterial adherence was investigated by incubating in human fresh bile adding E. coli and Enterobacter at 37±1 ℃ . Scanning electron microscopy （SEM） and fluorescence staining were used for observing bacterial colonization and biofilm formation. The results show that chitosan film was uniformly deposited on material surface, and the composition of the film did not change through cross-linking, but the crystallinity of chitosan film become well. Comparing to un-modified sample, the E. coli and Enterococcus adhesion amount and colonization on the surface of modified sample were significantly decreased by fluorescence staining and SEM. It is suggested that chitosan could be applied to biliary stent in clinical because of its antimicrobial activities.

A nanoplatform with oxygen self-supplying and heat-sensitizing capabilities enhances the efficacy of photodynamic therapy in eradicating multidrug-resistant biofilms

Bacterial biofilms,especially those caused by multidrug-resistant bacteria,have emerged as one of the greatest dangers to global public health.The acceleration of antimicrobial resistance to conventional an-tibiotics and the severe lack of new drugs necessitates the development of novel agents for biofilm eradication.Photodynamic therapy(PDT)is a promising non-antibiotic method for treating bacterial infections.However,its application in biofilm eradication is hampered by the hypoxic microenvironment of biofilms and the physical protection of extracellular polymeric substances.In this study,we develop a composite nanoplatform with oxygen(O_(2))self-supplying and heat-sensitizing capabilities to improve the PDT efficacy against biofilms.CaO_(2)/ICG@PDA nanoparticles(CIP NPs)are fabricated by combining calcium peroxide(CaO_(2))with the photosensitizer indocyanine green(ICG)via electrostatic interactions,followed by coating with polydopamine(PDA).The CIP NPs can gradually generate O_(2)in response to the acidic microenvironment of the biofilm,thereby alleviating its hypoxic state.Under near-infrared(NIR)irradiation,the nanoplatform converts O_(2)into a significant amount of singlet oxygen(^(1)O_(2))and heat to eradicate biofilm.The generated heat enhances the release of O_(2),accelerates the generation of^(1)O_(2)in PDT,increases cell membrane permeability,and increases bacterial sensitivity to^(1)O_(2).This nanoplatform significantly improves the efficacy of PDT in eradicating biofilm-dwelling bacteria without fostering drug resistance.Experiments on biofilm eradication demonstrate that this nanoplatform can eradicate over 99.9999%of methicillin-resistant Staphylococcus aureus(MRSA)biofilms under 5-min NIR irradiation.Notably,these integrated advantages enable the system to promote the healing of MRSA biofilm-infected wounds with negligible toxicity in vivo,indicating great promise for overcoming the obstacles associated with bacterial biofilm eradication.

What’s old is new again:Insights into diabetic foot microbiome

Diabetes is a chronic disease that is considered one of the most stubborn global health problems that continues to defy the efforts of scientists and physicians.The prevalence of diabetes in the global population continues to grow to alarming levels year after year,causing an increase in the incidence of diabetes complications and health care costs all over the world.One major complication of diabetes is the high susceptibility to infections especially in the lower limbs due to the immunocompromised state of diabetic patients,which is considered a definitive factor in all cases.Diabetic foot infections continue to be one of the most common infections in diabetic patients that are associated with a high risk of serious complications such as bone infection,limb amputations,and life-threatening systemic infections.In this review,we discussed the circumstances associated with the high risk of infection in diabetic patients as well as some of the most commonly isolated pathogens from diabetic foot infections and the related virulence behavior.In addition,we shed light on the different treatment strategies that aim at eradicating the infection.

Transcriptomic analysis of biofilm formation by Bacillus cereus under different carbon source conditions

Background:Previous studies found differences in the utilization of different carbon sources during biofilm formation by Bacillus cereus.Illumina HiSeq high-throughput sequencing technology was used to investigate the changes in gene transcript levels in Bacillus cereus biofilm bacteria under different carbon source conditions.Results:Compared with the control group,the number of differentially expressed genes in the glucose,maltose,lactose,and skim milksupplemented groups was 351,1136,133,and 487,respectively.The results showed that the pathways involved in the differentially expressed genes were mainly distributed in glycolysis and pentose phosphate pathway,tricarboxylic acid cycle,amino acid metabolism,and fatty acid metabolism.The gene expression of enzymes related to acetoin synthesis from pyruvate was mostly upregulated in the glucose-supplemented group.The gene expression of enzymes related to pyruvate synthesis of branched-chain amino acids in the maltose-supplemented group was mostly upregulated.In the lactose-supplemented group,the gene expression of acetoin biosynthesis from pyruvate was upregulated.Pyruvate production through glycolysis pathway increased in the skim milk-supplemented group,but the metabolic capacity of the tricarboxylic acid cycle did not change significantly.Conclusion:The content of pyruvate stored by Bacillus cereus biofilm bacteria through glycolysis or pentose phosphate pathway increased,but the carbon flux into the tricarboxylic acid cycle did not increase,which suggested that carbon fluxes in the extracellular polysaccharide synthesis pathway of the biofilm may be increased,resulting in increased biofilm biomass formation.

Molecular dynamics simulations of the interaction between OH radicals in plasma with poly-β-1–6-N-acetylglucosamine

Cold atmospheric plasma shows a satisfactory ability to inactivate bacterial biofilms that are difficult to remove using conventional methods in some cases. However, the researches on the inactivation mechanism are not quite sufficient. Poly-β-1–6-N-acetylglucosamine(PNAG),which is one of the important components in some biofilms, was used as the research subject,and the related mechanism of action triggered by different concentrations of the OH in plasma was studied using reactive molecular dynamics simulations. The results showed that OH radicals could not only trigger the hydrogen abstraction reaction leading to cleavage of the PNAG molecular structure, but undergo an OH addition reaction with PNAG molecules. New reaction pathways appeared in the simulations as the OH concentration increased, but the reaction efficiency first increased and then decreased. The simulation study in this paper could, to some extent, help elucidate the microscopic mechanism of the interaction between OH radicals in plasma and bacterial biofilms at the atomic level.

Transcriptomic analysis of biofilm formation by Bacillus cereus under different carbon source conditions

Background:Previous studies found differences in the utilization of different carbon sources during biofilm formation by Bacillus cereus.Illumina HiSeq high-throughput sequencing technology was used to investigate the changes in gene transcript levels in Bacillus cereus biofilm bacteria under different carbon source conditions.Results:Compared with the control group,the number of differentially expressed genes in the glucose,maltose,lactose,and skim milksupplemented groups was 351,1136,133,and 487,respectively.The results showed that the pathways involved in the differentially expressed genes were mainly distributed in glycolysis and pentose phosphate pathway,tricarboxylic acid cycle,amino acid metabolism,and fatty acid metabolism.The gene expression of enzymes related to acetoin synthesis from pyruvate was mostly upregulated in the glucose-supplemented group.The gene expression of enzymes related to pyruvate synthesis of branched-chain amino acids in the maltose-supplemented group was mostly upregulated.In the lactose-supplemented group,the gene expression of acetoin biosynthesis from pyruvate was upregulated.Pyruvate production through glycolysis pathway increased in the skim milk-supplemented group,but the metabolic capacity of the tricarboxylic acid cycle did not change significantly.Conclusion:The content of pyruvate stored by Bacillus cereus biofilm bacteria through glycolysis or pentose phosphate pathway increased,but the carbon flux into the tricarboxylic acid cycle did not increase,which suggested that carbon fluxes in the extracellular polysaccharide synthesis pathway of the biofilm may be increased,resulting in increased biofilm biomass formation.

New strategy for reversing biofilm-associated antibiotic resistance through ferrocene-substituted carborane ruthenium(Ⅱ)-arene complex

Bacterial biofilms are inherently resistant to antimicrobial agents and are difficult to eradicate with conventional antimicrobial agents, resulting in many persistent and chronic bacterial infections. In this contribution, a new strategy for reversing the biofilm-associated antibiotic resistance has been explored by induction of a carborane ruthenium(II)-arene complex (FcRuSB). Our results demonstrate that the FcRuSB could be utilized as an inducer to efficiently reverse the biofilm-associated antibiotic resistance of multidrug-resistant (MDR) clinical isolates of Staphylococcus aureus and Pseudomonas aeruginosa. The induced effect of FcRuSB is correlated with a considerable decrease in the expression of extracellular matrix proteins (EMP) of the two strains. The considerable decrease of the EMP of induced cells, resulting in the reduction of adherence and biofilm formation ability of the two types of MDR pathogens, and then can cause significantly enhanced sensitivity of them to antibiotics.

Immersed Boundary Approach to Biofilm Spread on Surfaces

We propose a computational model to study the growth and spread of bacterial biofilms on interfaces,as well as the action of antibiotics on them.Bacterial membranes are represented by boundaries immersed in a fluid matrix and subject to interaction forces.Growth,division and death of bacterial cells follow dynamic energy budget rules,in response to variations in environmental concentrations of nutrients,toxicants and substances released by the cells.In this way,we create,destroy and enlarge boundaries,either spherical or rod-like.Appropriate forces represent details of the interaction between cells,and the interaction with the environment.We can investigate geometrical arrangements and the formation of porous structures.Numerical simulations illustrate the evolution of top views and diametral slices of small biofilm seeds,as well as the action of antibiotics.We show that cocktails of antibiotics targeting active and dormant cells can entirely eradicate a biofilm.

Biofilm-overproducing Bacillus subtilis B12ΔYwcc decreases Cd uptake in Chinese cabbage through increasing Cd-immobilizing related gene abundance and root surface colonization

Biofilm-producing bacteria can decrease Cd uptake in vegetables, but mechanisms underlying this effect are poorly characterized. In this study, two mutant strains B12ΔYwcc and B12ΔSlr R were constructed from a biofilm-producing Bacillus subtilis strain B12. Then, the impacts of strain B12 and its high biofilm-producing mutant strain B12ΔYwcc and low biofilmproducing mutant strain B12ΔSlr R on Cd availability and uptake in Chinese cabbage and the related mechanisms were investigated in the Cd-polluted soil. Strain B12 and its mutants B12ΔYwcc and B12ΔSlr R increased the dry biomasses of edible tissues by 54%–130% compared with the controls. Strain B12 and its mutant B12ΔYwcc reduced the soil available Cd content by 36%–50% and root and edible tissue Cd contents by 23%–50% compared with the controls. Furthermore, the mutant strain B12ΔYwcc reduced the edible tissue Cd content by40% and increased the polysaccharide content by 23%, invertase activity by 139%, and gene copies of the cum A by 4.5-fold, eps A by 7.1-fold, and cad A by 4.3-fold, which were involved in Cd adsorption in the rhizosphere soils, respectively, compared with strain B12. The polysaccharide content and cum A, eps A, and cad A gene copy numbers showed significantly reverse correlations with the available Cd content. Notably, the mutant strain B12ΔYwcc showed better ability to colonize the vegetable root surface than strain B12. These findings demonstrated that the biofilm-overproducing mutant strain B12ΔYwcc increased the polysaccharide production and Cd-immobilizing related cum A, eps A, and cad A gene copies, resulting in lower Cd availability and accumulation in Chinese cabbage in the Cd-polluted soil.