Bacterial Biofilm Development Research Articles

The effect of 3D-printed bone tissue scaffolds geometrical designs on bacterial biofilm formation

Bone fractures are recognized as a global health problem. A common strategy to tackle this issue is to employ a tissue engineering scaffold to accelerate tissue healing. However, one of the main challenges that can result in delaying the recovery is the risk of bacterial infections. This study aims to assess the impact of the geometry and the porosity of tissue scaffolds on the Staphylococcus aureus biofilm formation. Three triply periodic minimal surface designs of Schwarz primitive (SP), gyroid (GY), and Schwarz diamond (SD) and re-entrant auxetic (RE) design were examined and compared to a reference design (RD) considering two different porosity levels of 75% and 45%. The amount of biofilm was quantified using crystal violet assay and was visualized using scanning electron microscopy. The SP scaffold, with low porosity, exhibited a significantly less amount of bacterial biofilm formation and was regarded as having the best design among the others, while the SD with low porosity showed the greatest amount of biofilm. The morphological analysis was also in line with the crystal violet assay results. On the other hand, the surface roughness was affected by the complexity, geometrical variations, and limitations of fused filament fabrication three-dimensional printing. For the RD, SP, GY, and SD designs, an increase in surface roughness was demonstrated to increase the production of bacterial biofilms. Without statistical significance, the RE design showed the opposite trend. Contrary to other designs, the increase in pore size of the SP and GY designs was associated with the development of bacterial biofilms. This study suggests that it is possible to minimize the likelihood of bacterial biofilm formation by optimizing the scaffold geometry and its manufacturing.

N-АЦЕТИЛЦИСТЕИН: ВОЗМОЖНОСТИ И ПЕРСПЕКТИВЫ ПРИМЕНЕНИЯ ПРИ ИНФЕКЦИЯХ МОЧЕВЫВОДЯЩИХ ПУТЕЙ

A review of literature data on the prospects for the use of acetylcysteine in the treatment of urinary tract infections is presented. In addition to the well-known mucolytic effect, ACC has antibacterial, anti-inflammatory and antioxidant properties. A number of studies have revealed the ability of ACC to suppress the development of bacterial biofilms, which is important in the treatment of chronic infections, including urinary tract infections. Studies have also demonstrated the anti-inflammatory activity of ACC, expressed in a decrease in the production of interleukins. Of interest is the property of ACC to suppress urease Proteus mirabilis, one of the main causative agents of nasocomial infection in urology. This property of ACC can be used in the prevention of infections associated with urinary catheters and ureteral stents incrustations.

A versatile micromodel technology to explore biofilm development in porous media flows.

Bacterial biofilms that grow in porous media are critical to ecosystem processes and applications ranging from soil bioremediation to bioreactors for treating wastewater or producing value-added products. However, understanding and engineering the complex phenomena that drive the development of biofilms in such systems remains a challenge. Here we present a novel micromodel technology to explore bacterial biofilm development in porous media flows. The technology consists of a set of modules that can be combined as required for any given experiment and conveniently tuned for specific requirements. The core module is a 3D-printed micromodel where biofilm is grown into a perfusable porous substrate. High-precision additive manufacturing, in particular stereolithography, is used to fabricate porous scaffolds with precisely controlled architectures integrating flow channels with diameters down to several hundreds of micrometers. The system is instrumented with: ultraviolet-C light-emitting diodes; on-line measurements of oxygen consumption and pressure drop across the porous medium; camera and spectrophotometric cells for the detection of biofilm detachment events at the outlet. We demonstrate how this technology can be used to study the development of Pseudomonas aeruginosa biofilm for several days within a network of flow channels. We find complex dynamics whereby oxygen consumption reaches a steady-state but not the pressure drop, which instead features a permanent regime with large fluctuations. We further use X-ray computed microtomography to image the spatial distribution of biofilms and computational fluid dynamics to link biofilm development with local flow properties. By combining the advantages of additive manufacturing for the creation of reproducible 3D porous microarchitectures with the flow control and instrumentation accuracy of microfluidics, our system provides a platform to study the dynamics of biofilm development in 3D porous media and to rapidly test new concepts in process engineering.

Review on Anti-Biofilm Potential of CRISPR-Cas System on Various Pathogens

The development of bacterial biofilms entails regulatory as well as signalling mechanisms that regulate the shift from a mobile to astationary state of existence, the production of the extracellular polymeric matrix, and the progression of 3-dimensional biofilm formation withemphasis on how easily they may be created and how crucial they are in biological, ecological, and industrial settings, biofilms are the subject ofextensive research. Biofilms and a variety of pathogenic human disorders are frequently linked. Since bacteria in biofilms can resist antibiotics, theimmune system, and other treatments, biofilm infections are typically long-lasting. Many prokaryotes include CRISPR-associated proteins (Cas), a solidadaptable immunological system that may be programmed to damage the bacterial genomes and induce cell death. Short palindromic repeats that aregrouped and adequately spaced together make up CRISPR-Cas. In light of this, CRISPR-Cas can be seen as an exciting strategy to address andovercome antibiotic resistance. Furthermore, the CRISPR-Cas system can create "precise antimicrobials" that target bacterial infections according tospecific DNA sequences. This CRISPR-Cas technique is susceptible to drug-resistant microorganisms due to its selective targeting of the genesinvolved in biofilm formation, pathogenicity, and antibiotic resistance. However, this method requires potent vectors for the CRISPR-Cas system toaccess the bacterial genomes. As vectors, genetically engineered Phage, liposomes, and lipid-mediated nanoparticles are exciting options. Thistechnique has been used to prevent extracellular and intracellular from forming biofilms. The most current developments in creating innovations andpossible advantages of the various CRISPR-Cas delivery methods for the deliberate eradication of bacterial pathogens will be covered in this review,focussing mainly on the anti-biofilm potential, which is found to be one of the primary causes of the difficulty of irradiation of Multi-drug resistantbacteria. Additionally, each distribution system's positive aspects are highlighted, along with its challenges and potential for advancement in the future.

Ceftazidime-assisted synthesis of ultrasmall chitosan nanoparticles for biofilm penetration and eradication of Pseudomonas aeruginosa

Pseudomonas aeruginosa (P. aeruginosa) infections present a grave threat to immunocompromised individuals, particularly those with cystic fibrosis due to the development of bacterial biofilms. In this study, we engineered self-assembling chitosan-ceftazidime nanoparticles (CSCE) capable of effectively penetrating biofilms and eradicating P. aeruginosa. The CSCE nanoparticles were synthesized through ionic cross-linking, combining negatively charged ceftazidime with positively charged chitosan, resulting in uniform nanoparticles measuring approximately 40 nm in diameter, exhibiting high dispersity and excellent biocompatibility. Remarkably, these nanoparticles exhibited significant inhibition of P. aeruginosa growth, reduced pyocyanin production, and diminished biofilm formation, achieving a maximum inhibition rate of 22.44%. Furthermore, in vivo investigations demonstrated enhanced survival in mice with abdominal P. aeruginosa infection following treatment with CSCE nanoparticles, accompanied by reduced levels of inflammatory cytokines Interleukin-6 (125.79 ± 18.63 pg/mL), Interleukin-17 (125.67 ± 5.94 pg/mL), and Tumor Necrosis Factor-α (135.4 ± 11.77 pg/mL). Critically, mice treated with CSCE nanoparticles showed no presence of bacteria in the bloodstream following intraperitoneal P. aeruginosa infection. Collectively, our findings highlight the potential of these synthesized nanoparticles as effective agents against P. aeruginosa infections.

Microfluidics for Biofilm Studies

Biofilms are multicellular communities held together by a self-produced extracellular matrix and exhibit a set of properties that distinguish them from free-living bacteria. Biofilms are exposed to a variety of mechanical and chemical cues resulting from fluid motion and mass transport. Microfluidics provides the precise control of hydrodynamic and physicochemical microenvironments to study biofilms in general. In this review, we summarize the recent progress made in microfluidics-based biofilm research, including understanding the mechanism of bacterial adhesion and biofilm development, assessment of antifouling and antimicrobial properties, development of advanced in vitro infection models, and advancement in methods to characterize biofilms. Finally, we provide a perspective on the future direction of microfluidics-assisted biofilm research.

Chromatography-Free Synthesis of β-Carboline 1-Hydrazides and an Investigation of the Mechanism of Their Bioactivity: The Discovery of β-Carbolines as Promising Antifungal and Antibacterial Candidates.

Agrochemical science prioritizes the discovery and effective synthesis of innovative and promising lead compounds. Herein, we developed an efficient column chromatography-free synthesis for β-carboline 1-hydrazides via a mild CuBr2-catalyzed oxidation and investigated the antifungal and antibacterial activities and mechanisms for these compounds. In our study, compounds 4de (EC50 = 0.23 μg·mL-1) and 4dq (EC50 = 0.11 μg·mL-1) displayed the best efficacy, demonstrating enhancements in inhibitory activity of more than 20-fold against Ggt compared to silthiopham (EC50 = 2.39 μg·mL-1). Additionally, compound 4de (EC50 = 0.21 μg·mL-1) demonstrated outstanding in vitro antifungal activities as well as in vivo curative activities against Fg. According to preliminary mechanistic studies, β-carboline 1-hydrazides led to the accumulation of reactive oxygen species, destruction of cell membranes, and dysregulation of histone acetylation. Furthermore, several substances exhibited antibacterial activity against Psg and Cms by preventing the development of bacterial biofilms.

Preparation of antibacterial PHBV/GO nanocomposite membranes via electrospinning

Poly(3-hydroxybutyrate-co-3-hydroxypentanoate) (PHBV)/graphene oxide (GO) composite membranes were electrospun with an aim at fabricating antibacterial nanofibrous scaffolds for tissue engineering application. Randomly and aligned oriented PHBV/GO fibrous membrane were prepared by controlling the mixed solvent, GO concentration and process parameters, and their effect on the morphology and structure of the membranes was investigated. When 10 wt% GO nanosheets was incorporated into the composite spinning dope, PHBV/GO membrane presented a 18.7 MPa breaking strength and 5.51 × 102 MPa tensile modulus, much greater than 14.3 MPa and 1.76 × 102 MPa of PHBV membrane. Colonies forming units of E.coli and S.aureus reduced by 81.7% and 71.8% for randomly oriented PHBV/GO, and 84.2% and 73.4% for aligned oriented PHBV/GO when compared to PHBV membrane. The development of bacterial biofilm on PHBV/GO membrane reduced greatly when incorporating 5–10 wt% GO. The antibacterial PHBV/GO membranes with better mechanical properties showed great promise for further studies on bone tissue repair.

Fucoidan-Containing, Low-Adhesive Siloxane Coatings for Medical Applications: Inhibition of Bacterial Growth and Biofilm Development.

The deposition of low-adhesive siloxane coatings is a current trend for the non-toxic control of bacterial growth and biofilm formation. Total elimination of biofilm formation has not been reported so far. The aim of this investigation was to study the ability of a non-toxic, natural, biologically active substance, such as fucoidan, to inhibit bacterial growth on similar medical coatings. The fucoidan amount was varied, and its impact on the bioadhesion-influencing surface characteristics, as well as on bacterial cell growth, was investigated. The inclusion of up to 3-4 wt.% brown algae-derived fucoidan in the coatings increases their inhibitory effect, more significantly on the Gram-positive bacterium S. aureus than on the Gram-negative bacterium Escherichia coli. The biological activity of the studied siloxane coatings was ascribed to the formation of a low-adhesive, biologically active surface top layer consisting of siloxane oil and dispersed water-soluble fucoidan particles. This is the first report on the antibacterial activity of fucoidan-containing medical siloxane coatings. The experimental results give reason to expect that relevantly selected, natural biologically active substances can be efficient in the non-toxic control of bacterial growth on medical devices and, as a result, medical device-associated infections.

Synergistic effects of biofilm-producing PGPR strains on wheat plant colonization, growth and soil resilience under drought stress

Drought stress substantially impedes crop productivity throughout the world. Microbial based approaches have been considered a potential possibility and are under study. Based on our prior screening examination, two distinct and novel biofilm-forming PGPR strains namely Bacillus subtilis-FAB1 and Pseudomonas azotoformans-FAP3 are encompassed in this research. Bacterial biofilm development on glass surface, microtiter plate and seedling roots were assessed and characterized quantitatively and qualitatively by light and scanning electron microscopy. Above two isolates were further evaluated for their consistent performance by inoculating on wheat plants in a pot-soil system under water stresses. Bacterial moderate tolerance to ten-day drought was recorded on the application of individual strains with wheat plants; however, the FAB1 + FAP3 consortium expressively improved wheat survival during drought. The strains FAB1 and FAP3 displayed distinct and multifunctional plant growth stimulating attributes as well as effective roots and rhizosphere colonization in combination which could provide sustained wheat growth during drought. FAB1 and FAP3-induced alterations cooperatively conferred improved plant drought tolerance by controlling physiological traits (gs, Ci, E, iWUE and PN), stress indicators (SOD, CAT, GR, proline and MDA content) and also maintained physico-chemical attributes and hydrolytic enzymes including DHA, urease, ALP, protease, ACP and β glucosidase in the soil. Our findings could support future efforts to enhance plant drought tolerance by engineering the rhizobacterial biofilms and associated attributes which requires in-depth exploration and exploiting potential native strains for local agricultural application.

Main approaches of biofilm formation in composite resins: a concise systematic review

Introduction: Composite resins are widely used in clinical dental practice to restore the structure lost by caries and dental defects, achieving aesthetic and functional results. Failures in the photoactivation process can cause problems in the mechanical resistance to wear in regions of direct occlusal contact on the restorative material. This occurs due to the reduction in the degree of conversion of monomers into polymers, compromising the clinical performance of the composite resin. Polymerization shrinkage leads to microleakage of the restoration, leading to staining and the development of bacterial biofilms. Objective: It was to carry out a systematic review of the main scientific evidence of biofilm formation on composite resin veneers, identifying potential predictors, as well as proposals to mitigate this problem. Methods: The systematic review rules of the PRISMA Platform were followed. The search was carried out from December 2022 to March 2023 in the Scopus, PubMed, Science Direct, Scielo, and Google Scholar databases, using articles from 2015 to 2022. The quality of the studies was based on the GRADE instrument and the risk of bias was analyzed accordingly, according to the Cochrane instrument. Results and Conclusion: A total of 125 articles were found, 25 articles were evaluated and 20 were included and developed in this systematic review study. Considering the Cochrane tool for risk of bias, the overall assessment resulted in 10 studies with a high risk of bias and 70 studies that did not meet GRADE. It was concluded that the surface properties of resin-based composite materials, as well as surface treatments, can strongly affect bacterial adhesion and biofilm formation. In addition, scientific evidence highlights that cariogenic biofilm formation can alter the surface properties of materials, thus stimulating bacterial adhesion and biofilm formation. Photoactivation allows the curing and crosslinking of polymer chains, essential for the clinical longevity of the composite resin. Insufficient photoactivation culminates in marginal microleakage in restorations and biofilm accumulation on surfaces exposed to the oral environment.

Review of Antimicrobial Nanocoatings in Medicine and Dentistry: Mechanisms of Action, Biocompatibility Performance, Safety, and Benefits Compared to Antibiotics.

This review discusses topics relevant to the development of antimicrobial nanocoatings and nanoscale surface modifications for medical and dental applications. Nanomaterials have unique properties compared to their micro- and macro-scale counterparts and can be used to reduce or inhibit bacterial growth, surface colonization and biofilm development. Generally, nanocoatings exert their antimicrobial effects through biochemical reactions, production of reactive oxygen species or ionic release, while modified nanotopographies create a physically hostile surface for bacteria, killing cells via biomechanical damage. Nanocoatings may consist of metal nanoparticles including silver, copper, gold, zinc, titanium, and aluminum, while nonmetallic compounds used in nanocoatings may be carbon-based in the form of graphene or carbon nanotubes, or composed of silica or chitosan. Surface nanotopography can be modified by the inclusion of nanoprotrusions or black silicon. Two or more nanomaterials can be combined to form nanocomposites with distinct chemical or physical characteristics, allowing combination of different properties such as antimicrobial activity, biocompatibility, strength, and durability. Despite their wide range of applications in medical engineering, questions have been raised regarding potential toxicity and hazards. Current legal frameworks do not effectively regulate antimicrobial nanocoatings in matters of safety, with open questions remaining about risk analysis and occupational exposure limits not considering coating-based approaches. Bacterial resistance to nanomaterials is also a concern, especially where it may affect wider antimicrobial resistance. Nanocoatings have excellent potential for future use, but safe development of antimicrobials requires careful consideration of the "One Health" agenda, appropriate legislation, and risk assessment.

Staphylococcus aureus Behavior on Artificial Surfaces Mimicking Bone Environment

Infections, which interfere with bone regeneration, may be a critical issue to consider during the development of biomimetic material. Calcium phosphate (CaP) and type I collagen substrates, both suitable for bone-regeneration dedicated scaffolds, may favor bacterial adhesion. Staphylococcus aureus possesses adhesins that allow binding to CaP or collagen. After their adhesion, bacteria may develop structures highly tolerant to immune system attacks or antibiotic treatments: the biofilms. Thus, the choice of material used for scaffolds intended for bone sites is essential to provide devices with the ability to prevent bone and joint infections by limiting bacterial adhesion. In this study, we compared the adhesion of three different S. aureus strains (CIP 53.154, SH1000, and USA300) on collagen- and CaP-coating. Our objective was to evaluate the capacity of bacteria to adhere to these different bone-mimicking coated supports to better control the risk of infection. The three strains were able to adhere to CaP and collagen. The visible matrix components were more important on CaP- than on collagen-coating. However, this difference was not reflected in biofilm gene expression for which no change was observed between the two tested surfaces. Another objective was to evaluate these bone-mimicking coatings for the development of an in vitro model. Thus, CaP, collagen-coatings, and the titanium-mimicking prosthesis were simultaneously tested in the same bacterial culture. No significant differences were found compared to adhesion on surfaces independently tested. In conclusion, these coatings used as bone substitutes can easily be colonized by bacteria, especially CaP-coating, and must be used with an addition of antimicrobial molecules or strategies to avoid bacterial biofilm development.

A Comprehensive Mechanistic Antibacterial and Antibiofilm Study of Potential Bioactive ((BpA)2bp)Cu/Zn Complexes via Bactericidal Mechanisms against Escherichia coli.

Bacterial resistance to antibiotics and host defense systems is primarily due to bacterial biofilm formation in antibiotic therapy. In the present study, two complexes, bis (biphenyl acetate) bipyridine Cu (II) (1) and bis (biphenyl acetate) bipyridine Zn (II) (2), were tested for their ability to prevent biofilm formation. The minimum inhibitory concentration and minimum bactericidal concentration of complexes 1 and 2 were 46.87 ± 1.822 and 93.75 ± 1.345 and 47.87 ± 1.345 and 94.85 ± 1.466 μg/mL, respectively. The significant activity of both complexes was attributed to the damage caused at the membrane level and was confirmed using an imaging technique. The biofilm inhibitory potential levels of complexes 1 and 2 were 95% and 71%, respectively, while the biofilm eradication potential levels were 95% and 35%, respectively, for both complexes. Both the complexes showed good interactions with the E. coli DNA. Thus, complexes 1 and 2 are good antibiofilm agents that exert their bactericidal actions possibly by disrupting the bacterial membrane and interacting with the bacterial DNA, which can act as a powerful agent to restrain the development of bacterial biofilm on therapeutic implants.

Nature‐Inspired Biomimetic Surfaces for Controlling Bacterial Attachment and Biofilm Development

AbstractThe use of antibacterial and antifouling materials is widely being investigated to combat the increasing risk associated with bacterial infections and the evolution of drug‐resistant bacteria. Efficient antibacterial materials can be fabricated by mimicking the topography found on the surface of natural antibacterial materials. Natural materials such as the wings of cicadas and dragonflies have evolved to use the structural features on their surface to attain bactericidal properties. The nanopillars/nanospikes present on these natural materials physically damage the bacterial cells that settle on the nanostructures resulting in cell lysis and death. This article reviews the role of nanostructures found on the surface of some of these natural antibacterial and antifouling materials such as lotus leaf, cicadas and dragonflies wings, shark skin, and rose petals. These natural structures provide guidelines for the design of synthetic bio‐inspired materials. This review article also presents some novel fabrication techniques used to produce biomimetic micro‐ and nano‐structures on synthetic material surfaces. The role of size, shape, aspect ratio, and spacing between the micro/nano‐structures on the bactericidal properties is also discussed. Finally, the review is finished with the author's view on the future of the field.

Facing Resistant Bacteria with Plant Essential Oils: Reviewing the Oregano Case.

Antibiotic resistance is a serious global threat, and the misuse of antibiotics is considered its main cause. It is characterized by the expression of bacterial defense mechanisms, e.g., β-lactamases, expulsion pumps, and biofilm development. Acinetobacter baumannii and Pseudomonas aeruginosa are antibiotic-resistant species that cause high morbidity and mortality. Several alternatives are proposed to defeat antibiotic resistance, including antimicrobial peptides, bacteriophages, and plant compounds. Terpenes from different plant essential oils have proven antimicrobial action against pathogenic bacteria, and evidence is being generated about their effect against antibiotic-resistant species. That is the case for oregano essential oil (Lippia graveolens), whose antibacterial effect is widely attributed to carvacrol, its main component; however, minor constituents could have an important contribution. The analyzed evidence reveals that most antibacterial evaluations have been performed on single species; however, it is necessary to analyze their activity against multispecies systems. Hence, another alternative is using plant compounds to inactivate hydrolytic enzymes and biofilms to potentiate antibiotics' effects. Despite the promising results of plant terpenes, more extensive and deep mechanistic studies are needed involving antibiotic-resistant multispecies to understand their full potential against this problem.

Current Status of Preventive and Therapeutic Strategies Against Biofilm Formation in Arthroplasty

Total joint replacements have increased significantly, resulting in a corresponding increase in infections. Consequently, patients may undergo additional surgical procedures and be prescribed antibiotics for a prolonged period due to these infections. Periprosthetic joint infections are associated with the development of bacterial biofilms. The biofilm is a microbial community attached to a surface containing one or more bacterial species. In orthopedics, biofilm-forming bacteria are the most severe infection that can lead to multiple operations, prolonged antibiotic therapy, morbidity, and increased health-care expenditures. These biofilm communities pose several clinical challenges relating to infection prevention, detection, and treatment. Over the past few years, biofilm formation mechanisms have been extensively studied, as have the mechanisms by which bacteria communicate within biofilms to perform specialized functions, such as persister cells. Currently, the orthopedic literature is very scarce, and understanding the cause and eradicating the disease requires a deep understanding. Several studies have demonstrated that the delivery of antibiotics locally through absorbable carriers and novel coatings for prostheses can deliver high concentrations of antibiotics. This literature review aims to identify mechanisms and structures of biofilm, especially in the context of arthroplasty, and to provide strategic guidance on current diagnosis, prevention, and target-specific treatment. In addition, the review discusses future diagnostic and therapeutic advancements.

Antibiofilm Activity of Biocide Metal Ions Containing Bioactive Glasses (BGs): A Mini Review.

One of the major clinical issues during the implantation procedure is the bacterial infections linked to biofilms. Due to their tissue localization and the type of bacteria involved, bacterial infections at implant sites are usually difficult to treat, which increases patient morbidity and even mortality. The difficulty of treating biofilm-associated infections and the emergence of multidrug-resistant bacteria are further challenges for the scientific community to develop novel biomaterials with excellent biocompatibility and antibacterial properties. Given their ability to stimulate bone formation and have antibacterial properties, metal ion-doped bioactive glasses (BGs) have received considerable research. This mini review aims to be successful in presenting the developments made about the role of biocide metal ions incorporated into BGs against the development of bacterial biofilms and the spread of nosocomial diseases.

Assessing in vivo and in vitro biofilm development by Streptococcus dysgalactiae subsp. dysgalactiae using a murine model of catheter-associated biofilm and human keratinocyte cell.

Streptococcus dysgalactiae subsp. dysgalactiae (SDSD) is an important agent of bovine mastitis. This infection causes an inflammatory reaction in udder tissue, being the most important disease-causing significant impact on the dairy industry. Therefore, it leads to an increase in dairy farming to meet commercial demands. As a result, there is a major impact on both the dairy industry and the environment including global warming. Recurrent mastitis is often attributed to the development of bacterial biofilms, which promote survival of sessile cells in hostile environments, and resistance to the immune system defense and antimicrobial therapy. Recently, we described the in vitro biofilm development on abiotic surfaces by bovine SDSD. In that work we integrated microbiology, imaging, and computational methods to evaluate the biofilm production capability of SDSD isolates on abiotic surfaces. Additionally, we reported that bovine SDSD can adhere and internalize human cells, including human epidermal keratinocyte (HEK) cells. We showed that the adherence and internalization rates of bovine SDSD isolates in HEK cells are higher than those of a SDSD DB49998-05 isolated from humans. In vivo, bovine SDSD can cause invasive infections leading to zebrafish morbidity and mortality. In the present work, we investigated for the first time the capability of bovine SDSD to develop biofilm in vivo using a murine animal model and ex-vivo on human HEK cells. Bovine SDSD isolates were selected based on their ability to form weak, moderate, or strong biofilms on glass surfaces. Our results showed that SDSD isolates displayed an increased ability to form biofilms on the surface of catheters implanted in mice when compared to in vitro biofilm formation on abiotic surface. A greater ability to form biofilm in vitro after animal passage was observed for the VSD45 isolate, but not for the other isolates tested. Besides that, in vitro scanning electron microscopy demonstrated that SDSD biofilm development was visible after 4 hours of SDSD adhesion to HEK cells. Cell viability tests showed an important reduction in the number of HEK cells after the formation of SDSD biofilms. In this study, the expression of genes encoding BrpA-like (biofilm regulatory protein), FbpA (fibronectin-binding protein A), HtrA (serine protease), and SagA (streptolysin S precursor) was higher for biofilm grown in vivo than in vitro, suggesting a potential role for these virulence determinants in the biofilm-development, host colonization, and SDSD infections. Taken together, these results demonstrate that SDSD can develop biofilms in vivo and on the surface of HEK cells causing important cellular damages. As SDSD infections are considered zoonotic diseases, our data contribute to a better understanding of the role of biofilm accumulation during SDSD colonization and pathogenesis not only in bovine mastitis, but they also shed some lights on the mechanisms of prosthesis-associated infection and cellulitis caused by SDSD in humans, as well.

Biofilm inhibition and bacterial eradication by C-dots derived from polyethyleneimine-citric acid

Protonable polyethyleneimine (PEI) and citric acid (CA) based C-dots were prepared via a hydrothermal process, yielding particles with a hydrodynamic diameter of ~100 and ~80 nm, and zeta potential of − 2.3 ± 0.1 and + 23.4 ± 0.2 mV for PEI-CA C-dots and their protonated form, respectively. Treating Staphylococcus aureus with these C-dots resulted in a statistically significant reduction in bacterial growth, specifically growth in the planktonic phase as well as in the development of bacterial biofilm when compared to untreated (p < 0.05). When 24 h matured S. aureus biofilms were treated with C-dots, a significant disruption and dispersion of bacteria from the existing biofilms was noted as compared to untreated (p < 0.05). Other Gram-positive microorganisms (B. cereus, S. epidermidis, S. pyogenes) and Gram-negative (P. mirabilis, E. coli, and P. aeruginosa) were also susceptible to the antimicrobial activity of the PEI-CA C-dots with significant inhibition of bacterial growth noted for all organisms after C-dot treatment. Only for B. cereus, S. epidermidis, and P. aeruginosa was this reduction in total growth reflected in decreased planktonic growth. However, biofilm formation by all organisms was reduced significantly upon treatment with the C-dots, including those for which planktonic growth was not impacted.