@article {pmid34570460,
year = {2021},
author = {Tan, GR and Hsu, CS and Zhang, Y},
title = {pH-Responsive Hybrid Nanoparticles for Imaging Spatiotemporal pH Changes in Biofilm-Dentin Microenvironments.},
journal = {ACS applied materials & interfaces},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsami.1c11162},
pmid = {34570460},
issn = {1944-8252},
abstract = {Engineering highly sensitive nanomaterials to monitor spatiotemporal pH changes has rather broad applications in studying various biological systems. Intraoral/biofilm-tooth pH is the single parameter that has demonstrated accurate assessment of dental caries risk, reflecting the summative integrated outcome of the complicated interactions between three etiological factors, namely, microorganisms/biofilm, diet/carbohydrates, and tooth/saliva/host. However, there is little to no technology/system capable of accurately probing simultaneously both the micro-pH profiles in dentin tissues and acidogenic oral biofilms and examining the pathophysiologic acid attacks with high spatial/temporal resolution. Therefore, a highly sensitive pH-responsive hybrid nanoparticle (pH-NP) is developed and coupled with an ex vivo tooth-biofilm caries model to simulate and study the key cariogenic determinants/steps. The pH-NP emits two distinct fluorescences with mutually inversely proportional intensities that vary accordingly to the proximity pH and with a ratiometric output sensitivity of 13.4-fold across a broad clinically relevant pH range of 3.0-8.0. Using [H+], in addition to pH, to calculate the "area-under-curve" corroborates the "minimum-pH" in semiquantifying the demineralizing potential in each biofilm-dentin zones/depth. The data mechanistically elucidates a two-pronged cariogenic effect of a popular-acidic-sweet-drink, in inundating the biofilm/tooth-system with H+ ions from both the drink and the metabolic byproducts of the biofilm.},
}

@article {pmid34569158,
year = {2021},
author = {Javadiyan, S and Cooksley, CM and Bouras, GS and Kao, SS and Bennett, CA and Wormald, PJ and Vreugde, S and Psaltis, AJ},
title = {Investigation of Kappa Carrageenan's muco-adhesive, antibacterial, and anti-biofilm properties.},
journal = {International forum of allergy & rhinology},
volume = {},
number = {},
pages = {},
doi = {10.1002/alr.22899},
pmid = {34569158},
issn = {2042-6984},
}

@article {pmid34567189,
year = {2021},
author = {Yousefpour, Z and Davarzani, F and Owlia, P},
title = {Evaluating of the Effects of Sub-MIC Concentrations of Gentamicin on Biofilm Formation in Clinical Isolates of Pseudomonas aeruginosa.},
journal = {Iranian journal of pathology},
volume = {16},
number = {4},
pages = {403-410},
doi = {10.30699/IJP.20201.524220.2584},
pmid = {34567189},
issn = {1735-5303},
abstract = {Background & Objective: The ability of Pseudomonas aeruginosa to form biofilm has an important role in establishment of chronic phase of infections. Biofilm formation can be affected by antibiotics sub-MIC concentrations. The principal aim of the present study was to evaluate the effect of gentamicin at sub-MIC concentrations on biofilm formation in 100 Pseudomonas aeruginosa clinical isolates.

Methods: Determination of minimal inhibitory concentration of gentamicin for clinical isolates was done using micro broth dilution method. The amount of biofilm formation in the treated and untreated isolates with gentamicin sub-MIC (1/2&1/4MIC) concentrations was evaluated using microtitre plate assay. pelA and pslA genes were detected in clinical isolates by PCR method.

Results: 99% of clinical isolates were biofilm producer. Different changes in amount of biofilm formation were observed in the treated clinical isolates with sub-MIC concentrations of gentamicin. Two dominant changes were observed in 80% of clinical isolates. These concentrations had inhibitory effect on biofilm formation in 46.4% of isolates and caused a significant decrease in its amount. While in 31.3% of the isolates, the biofilm formation was significantly increased. The frequency of pelA and pslA genes among clinical isolates was 100%.

Conclusion: gentamicin sub-MIC concentrations cause different changes on biofilm formation of Pseudomonas aeruginosa clinical isolates. Therefore, further studies are needed for discovering new treatment strategies and using sub-MIC concentrations of the antibiotic in prevention and treatment of Pseudomonas aeruginosa infections.},
}

@article {pmid34567162,
year = {2021},
author = {Memar, MY and Adibkia, K and Farajnia, S and Samadi Kafil, H and Khalili, Y and Azargun, R and Ghotaslou, R},
title = {In-vitro Effect of Imipenem, Fosfomycin, Colistin, and Gentamicin Combination against Carbapenem-resistant and Biofilm-forming Pseudomonas aeruginosa Isolated from Burn Patients.},
journal = {Iranian journal of pharmaceutical research : IJPR},
volume = {20},
number = {2},
pages = {286-296},
doi = {10.22037/ijpr.2020.111824.13380},
pmid = {34567162},
issn = {1735-0328},
abstract = {The aim of this study was to investigate in-vitro antibacterial and antibiofilm effect of colistin, imipenem, gentamicin, and fosfomycin alone and the various combinations against carbapenem-resistant Pseudomonas aeruginosa (P. aeruginosa). Eight carbapenem-resistant and biofilm-forming P. aeruginosa isolates from burn patients were collected. The mechanisms of resistance to carbapenem were determined by the phenotypic, PCR, and Real-Time PCR assays. The minimum inhibitory concentration (MIC) of antimicrobial agents was determined by the broth micro dilution. To detect any inhibitory effect of antibiotics against the biofilm, the biofilm inhibitory concentration was determined. To detect synergetic effects of the combinations of antibiotics, the checkerboard assay and the fractional inhibitory concentration (FIC) were used. The highest synergic effect was observed in colistin/fosfomycin and gentamicin/fosfomycin (5 of 8 isolates), and the lowest synergic effect was found in gentamicin/imipenem and colistin/gentamicin (1 of 8 isolates). Colistin/fosfomycin, imipenem/fosfomycin, colistin/imipenem, gentamicin/fosfomycin, and gentamicin/imipenem were shown synergic effect for 3, 2, 2, 2 and 1 isolates, respectively. The combination of antibiotics had different effects on biofilm and planktonic forms of P. aeruginosa. Therefore, a separate determination of inhibitory effects of the antibiotic in the combination is necessary. Fosfomycin/colistin and fosfomycin/gentamicin were more effective against planktonic form and fosfomycin/colistin against biofilm forms.},
}

@article {pmid34566936,
year = {2021},
author = {Poulin, MB and Kuperman, LL},
title = {Regulation of Biofilm Exopolysaccharide Production by Cyclic Di-Guanosine Monophosphate.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {730980},
doi = {10.3389/fmicb.2021.730980},
pmid = {34566936},
issn = {1664-302X},
abstract = {Many bacterial species in nature possess the ability to transition into a sessile lifestyle and aggregate into cohesive colonies, known as biofilms. Within a biofilm, bacterial cells are encapsulated within an extracellular polymeric substance (EPS) comprised of polysaccharides, proteins, nucleic acids, lipids, and other small molecules. The transition from planktonic growth to the biofilm lifecycle provides numerous benefits to bacteria, such as facilitating adherence to abiotic surfaces, evasion of a host immune system, and resistance to common antibiotics. As a result, biofilm-forming bacteria contribute to 65% of infections in humans, and substantially increase the energy and time required for treatment and recovery. Several biofilm specific exopolysaccharides, including cellulose, alginate, Pel polysaccharide, and poly-N-acetylglucosamine (PNAG), have been shown to play an important role in bacterial biofilm formation and their production is strongly correlated with pathogenicity and virulence. In many bacteria the biosynthetic machineries required for assembly of these exopolysaccharides are regulated by common signaling molecules, with the second messenger cyclic di-guanosine monophosphate (c-di-GMP) playing an especially important role in the post-translational activation of exopolysaccharide biosynthesis. Research on treatments of antibiotic-resistant and biofilm-forming bacteria through direct targeting of c-di-GMP signaling has shown promise, including peptide-based treatments that sequester intracellular c-di-GMP. In this review, we will examine the direct role c-di-GMP plays in the biosynthesis and export of biofilm exopolysaccharides with a focus on the mechanism of post-translational activation of these pathways, as well as describe novel approaches to inhibit biofilm formation through direct targeting of c-di-GMP.},
}

@article {pmid34566913,
year = {2021},
author = {Matar, GK and Ali, M and Bagchi, S and Nunes, S and Liu, WT and Saikaly, PE},
title = {Relative Importance of Stochastic Assembly Process of Membrane Biofilm Increased as Biofilm Aged.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {708531},
doi = {10.3389/fmicb.2021.708531},
pmid = {34566913},
issn = {1664-302X},
abstract = {The relative importance of different ecological processes controlling biofilm community assembly over time on membranes with different surface characteristics has never been investigated in membrane bioreactors (MBRs). In this study, five ultrafiltration hollow-fiber membranes - having identical nominal pore size (0.1μm) but different hydrophobic or hydrophilic surface characteristics - were operated simultaneously in the same MBR tank with a constant flux of 10 liters per square meter per hour (LMH). In parallel, membrane modules operated without permeate flux (0 LMH) were submerged in the same MBR tank, to investigate the passive microbial adsorption onto different hydrophobic or hydrophilic membranes. Samples from the membrane biofilm were collected after 1, 10, 20, and 30days of continuous filtration. The membrane biofilm microbiome were investigated using 16S rRNA gene amplicon sequencing from DNA and cDNA samples. Similar beta diversity trends were observed for both DNA- and cDNA-based analyses. Beta diversity analyses revealed that the nature of the membrane surface (i.e., hydrophobic vs. hydrophilic) did not seem to have an effect in shaping the bacterial community, and a similar biofilm microbiome evolved for all types of membranes. Similarly, membrane modules operated with and without permeate flux did not significantly influence alpha and beta diversity of the membrane biofilm. Nevertheless, different-aged membrane biofilm samples exhibited significant differences. Proteobacteria was the most dominant phylum in early-stage membrane biofilm after 1 and 10days of filtration. Subsequently, the relative reads abundance of the phyla Bacteroidetes and Firmicutes increased within the membrane biofilm communities after 20 and 30days of filtration, possibly due to successional steps that lead to the formation of a relatively aged biofilm. Our findings indicate distinct membrane biofilm assembly patterns with different-aged biofilm. Ecological null model analyses revealed that the assembly of early-stage biofilm community developed after 1 and 10days of filtration was mainly governed by homogenous selection. As the biofilm aged (days 20 and 30), stochastic processes (e.g., ecological drift) started to become important in shaping the assembly of biofilm community.},
}

@article {pmid34563667,
year = {2021},
author = {Galdiero, E and Ricciardelli, A and D'Angelo, C and de Alteriis, E and Maione, A and Albarano, L and Casillo, A and Corsaro, MM and Tutino, ML and Parrilli, E},
title = {Pentadecanoic acid against Candida albicans-Klebsiella pneumoniae biofilm: towards the development of an anti-biofilm coating to prevent polymicrobial infections.},
journal = {Research in microbiology},
volume = {},
number = {},
pages = {103880},
doi = {10.1016/j.resmic.2021.103880},
pmid = {34563667},
issn = {1769-7123},
abstract = {The ability to form biofilms is a common feature of microorganisms, which can colonize a variety of surfaces, such as host tissues and medical devices, resulting in infections highly resistant to conventional drugs. This aspect is particularly critical in polymicrobial biofilms involving both fungi and bacteria, therefore, to eradicate such severe infections, new and effective anti-biofilm strategies are needed. The efficacy of pentadecanal and pentadecanoic acid as anti-biofilm agents has been recently reported against different bacterial strains. Their chemical similarity with diffusible signal factors (DSFs), plus the already known ability of fatty acids to act as anti-biofilm agents, suggested to explore their use against Candida albicans and Klebsiella pneumoniae mixed biofilm. In this work, we demonstrated the ability of both molecules to prevent the formation and destabilize the structure of the dual-species biofilm. Moreover, the pentadecanoic acid anti-biofilm coating, previously developed through the adsorption of the fatty acid on polydimethylsiloxane, was proved to prevent the polymicrobial biofilm formation in dynamic conditions by confocal laser scanning microscopy analysis. Finally, the evaluation of the expression levels of some biofilm-related genes of Candida albicans and Klebsiella pneumoniae treated with pentadecanoic acid provided some insights into the molecular mechanisms underpinning its anti-biofilm effect.},
}

@article {pmid34563350,
year = {2021},
author = {Lin, WS and Alfaifi, AA and Gregory, RL},
title = {Response to Letter to the Editor regarding the article "Impact of caffeine on metabolic activity and biofilm formation of Candida albicans on acrylic denture resin in the presence of nicotine".},
journal = {The Journal of prosthetic dentistry},
volume = {},
number = {},
pages = {},
doi = {10.1016/j.prosdent.2021.08.017},
pmid = {34563350},
issn = {1097-6841},
}

@article {pmid34562786,
year = {2021},
author = {Wang, J and Zhu, J and Meng, J and Qiu, T and Wang, W and Wang, R and Liu, J},
title = {Baicalin inhibits biofilm formation by influencing primary adhesion and aggregation phases in Staphylococcus saprophyticus.},
journal = {Veterinary microbiology},
volume = {262},
number = {},
pages = {109242},
doi = {10.1016/j.vetmic.2021.109242},
pmid = {34562786},
issn = {1873-2542},
abstract = {The ability to form biofilms on surfaces makes Staphylococcus saprophyticus (S. saprophyticus) becomes the main pathogenic factor in nosocomial infections. Previously, we demonstrated that baicalin (Bac) inhibited azithromycin-resistant S. saprophyticus (ARSS) biofilm formation. This investigation aims to explore the influence of baicalin on primary adhesion and aggregation phases of biofilm formation, and the treatment effect of baicalin and azithromycin on ARSS biofilm-associated infection. Crystal violet (CV) staining and scanning electron microscope (SEM) observations clearly showed that sub-inhibitory concentration baicalin inhibited ARSS biofilm formation when baicalin was added before the adhesion and aggregation phases. Baicalin significantly increased the relative adhesion inhibition rate and decreased the rate of bacteria aggregation in a dose-dependent manner. Moreover, CLSM and cell lysis assays revealed that baicalin inhibited the production of surface proteins and cell autolysis in bacteria adhesion and aggregation phases of biofilm formation. Meanwhile, the relative expressions of adhesion-related and autolysis-related genes were down-regulated by baicalin. In vivo, the combination of baicalin and azithromycin succeeded in eradicating ARSS from the mouse cutaneous infection model and decreasing the pathological injuries, the expressions of cytokines in infected tissue, and the number of inflammatory cells in the blood. Simultaneously, baicalin decreased the bacterial burdens in tubes, the level of TNF-α, and the number of monocytes and neutrophils compared with that of the SS and azithromycin groups. Based on these results, baicalin inhibited the adhesion and aggregation phases of biofilm formation by influenced the production of surface proteins and cell autolysis. Baicalin and azithromycin synergetically treated ARSS biofilm-associated infection.},
}

@article {pmid34562533,
year = {2021},
author = {Zmejkoski, DZ and Zdravković, NM and Trišić, DD and Budimir, MD and Marković, ZM and Kozyrovska, NO and Todorović Marković, BM},
title = {Chronic wound dressings - Pathogenic bacteria anti-biofilm treatment with bacterial cellulose-chitosan polymer or bacterial cellulose-chitosan dots composite hydrogels.},
journal = {International journal of biological macromolecules},
volume = {191},
number = {},
pages = {315-323},
doi = {10.1016/j.ijbiomac.2021.09.118},
pmid = {34562533},
issn = {1879-0003},
abstract = {Since the pathogenic bacteria biofilms are involved in 70% of chronic infections and their resistance to antibiotics is increased, the research in this field requires new healing agents. New composite hydrogels were designed as potential chronic wound dressings composed of bacterial cellulose (BC) with chitosan polymer (Chi) - BC-Chi and chitosan nanoparticles (nChiD) - BC-nChiD. nChiD were obtained by gamma irradiation at doses: 20, 40 and 60 kGy. Physical and chemical analyses showed incorporation of Chi and encapsulation of nChiD into BC. The BC-Chi has the highest average surface roughness. BC-nChiD hydrogels show an irradiated dose-dependent increase of average surface roughness. New composite hydrogels are biocompatible with excellent anti-biofilm potential with up to 90% reduction of viable biofilm and up to 65% reduction of biofilm height. The BC-nChiD showed better dressing characteristics: higher porosity, higher wound fluid absorption and faster migration of cells (in vitro healing). All obtained results confirmed both composite hydrogels as promising chronic wound healing agents.},
}

@article {pmid34558030,
year = {2021},
author = {Wang, D and Bai, P and Zhang, B and Su, X and Jiang, X and Fang, T and Wang, J and Liu, C},
title = {Decreased biofilm formation in Proteus mirabilis after short-term exposure to a simulated microgravity environment.},
journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]},
volume = {},
number = {},
pages = {},
pmid = {34558030},
issn = {1678-4405},
abstract = {BACKGROUND: Microbes threaten human health in space exploration. Studies have shown that Proteus mirabilis has been found in human space habitats. In addition, the biological characteristics of P. mirabilis in space have been studied unconditionally. The simulated microgravity environment provides a platform for understanding the changes in the biological characteristics of P. mirabilis.

OBJECTIVE: This study intends to explore the effect of simulated microgravity on P. mirabilis, the formation of P. mirabilis biofilm, and its related mechanism.

METHODS: The strange deformable rods were cultured continuously for 14 days under microgravity simulated in high-aspect rotating vessels (HARVs). The morphology, growth rate, metabolism, and biofilm formation of the strain were measured, and the phenotypic changes of P. mirabilis were evaluated. Transcriptome sequencing was used to detect differentially expressed genes under simulated microgravity and compared with phenotype.

RESULTS: The growth rate, metabolic ability, and biofilm forming ability of P. mirabilis were lower than those of normal gravity culture under the condition of simulated microgravity. Further analysis showed that the decrease of growth rate, metabolic ability, and biofilm forming ability may be caused by the downregulation of related genes (pstS, sodB, and fumC).

CONCLUSION: The simulated microgravity condition enables us to explore the potential relationship between bacterial phenotype and molecular biology, thus opening up a suitable and constructive method for medical fields that have not been explored before. It provides a certain strategy for the treatment of P. mirabilis infectious diseases in space environment by exploring the microgravity of P. mirabilis.},
}

@article {pmid34558029,
year = {2021},
author = {Rather, MA and Gupta, K and Mandal, M},
title = {Microbial biofilm: formation, architecture, antibiotic resistance, and control strategies.},
journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]},
volume = {},
number = {},
pages = {},
pmid = {34558029},
issn = {1678-4405},
support = {BT/PR16149/NER/95/85/ 2015//Department of Biotechnology , Ministry of Science and Technology/ ; TU/Fin/R/18-19/339//Tezpur University/ ; },
abstract = {The assembly of microorganisms over a surface and their ability to develop resistance against available antibiotics are major concerns of interest. To survive against harsh environmental conditions including known antibiotics, the microorganisms form a unique structure, referred to as biofilm. The mechanism of biofilm formation is triggered and regulated by quorum sensing, hostile environmental conditions, nutrient availability, hydrodynamic conditions, cell-to-cell communication, signaling cascades, and secondary messengers. Antibiotic resistance, escape of microbes from the body's immune system, recalcitrant infections, biofilm-associated deaths, and food spoilage are some of the problems associated with microbial biofilms which pose a threat to humans, veterinary, and food processing sectors. In this review, we focus in detail on biofilm formation, its architecture, composition, genes and signaling cascades involved, and multifold antibiotic resistance exhibited by microorganisms dwelling within biofilms. We also highlight different physical, chemical, and biological biofilm control strategies including those based on plant products. So, this review aims at providing researchers the knowledge regarding recent advances on the mechanisms involved in biofilm formation at the molecular level as well as the emergent method used to get rid of antibiotic-resistant and life-threatening biofilms.},
}

@article {pmid34557273,
year = {2021},
author = {Ghaioumy, R and Tabatabaeifar, F and Mozafarinia, K and Mianroodi, AA and Isaei, E and Morones-Ramírez, JR and Afshari, SAK and Kalantar-Neyestanaki, D},
title = {Biofilm formation and molecular analysis of intercellular adhesion gene cluster (icaABCD) among Staphylococcus aureus strains isolated from children with adenoiditis.},
journal = {Iranian journal of microbiology},
volume = {13},
number = {4},
pages = {458-463},
pmid = {34557273},
issn = {2008-3289},
abstract = {Background and Objectives: It is well known that Staphylococcus aureus biofilm plays an important role in adenoiditis and biofilm resistance frequently results in failure of therapy. The goal of this study was to evaluate the biofilm production of S. aureus isolates obtained from adenoid specimens and assess the relationship between biofilm formation ability and ica operon genes.

Materials and Methods: A total of 112 adenoid samples were obtained from patients under 15 years old with adenoid hypertrophy. All S. aureus isolates were initially identified by standard microbiological tests and amplification of nuc by polymerase chain reaction (PCR) technique. Biofilm formation of S. aureus isolates was evaluated and icaADBC genes were detected by PCR technique.

Results: There were 46 isolates (41%) identified as S. aureus. The ability to produce biofilm was detected among total S. aureus isolates. Molecular study of ica operon revealed that 2 (6.3%) and 19 (59.4%) isolates carried icaA and icaD, respectively. The prevalence of icaA + icaD was seen among 11 (34.4%) S. aureus isolates, while icaC and icaB were not detected.

Conclusion: Our findings indicated that icaABCD operon are associated with biofilm formation in S. aureus isolates, however the absence of these genes may not necessarily exclude this property.},
}

@article {pmid34557173,
year = {2021},
author = {Franklin-Alming, FV and Kaspersen, H and Hetland, MAK and Bakksjø, RJ and Nesse, LL and Leangapichart, T and Löhr, IH and Telke, AA and Sunde, M},
title = {Exploring Klebsiella pneumoniae in Healthy Poultry Reveals High Genetic Diversity, Good Biofilm-Forming Abilities and Higher Prevalence in Turkeys Than Broilers.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {725414},
pmid = {34557173},
issn = {1664-302X},
abstract = {Klebsiella pneumoniae is a well-studied human pathogen for which antimicrobial resistant and hypervirulent clones have emerged globally. K. pneumoniae is also present in a variety of environmental niches, but currently there is a lack of knowledge on the occurrence and characteristics of K. pneumoniae from non-human sources. Certain environmental niches, e.g., animals, may be associated with high K. pneumoniae abundance, and these can constitute a reservoir for further transmission of strains and genetic elements. The aim of this study was to explore and characterize K. pneumoniae from healthy broilers and turkeys. A total of 511 cecal samples (broiler n = 356, turkey n = 155), included in the Norwegian monitoring program for antimicrobial resistance (AMR) in the veterinary sector (NORM-VET) in 2018, were screened for K. pneumoniae by culturing on SCAI agar. K. pneumoniae was detected in 207 (40.5%) samples. Among the broiler samples, 25.8% were positive for K. pneumoniae, in contrast to turkey with 74.2% positive samples (p < 0.01). Antibiotic susceptibility testing was performed, in addition to investigating biofilm production. Whole genome sequencing was performed on 203 K. pneumoniae isolates, and analysis was performed utilizing comparative genomics tools. The genomes grouped into 66 sequence types (STs), with ST35, ST4710 and ST37 being the most prevalent at 13.8%, 7.4%, and 5.4%, respectively. The overall AMR occurrence was low, with only 11.3% of the isolates showing both pheno- and genotypic resistance. Genes encoding aerobactin, salmochelin or yersiniabactin were detected in 47 (23.2%) genomes. Fifteen hypervirulent genomes belonging to ST4710 and isolated from turkey were identified. These all encoded the siderophore virulence loci iuc5 and iro5 on an IncF plasmid. Isolates from both poultry species displayed good biofilm-forming abilities with an average of OD595 0.69 and 0.64. To conclude, the occurrence of K. pneumoniae in turkey was significantly higher than in broiler, indicating that turkey might be an important zoonotic reservoir for K. pneumoniae compared to broilers. Furthermore, our results show a highly diverse K. pneumoniae population in poultry, low levels of antimicrobial resistance, good biofilm-forming abilities and a novel hypervirulent ST4710 clone circulating in the turkey population.},
}

@article {pmid34557170,
year = {2021},
author = {Lamret, F and Varin-Simon, J and Velard, F and Terryn, C and Mongaret, C and Colin, M and Gangloff, SC and Reffuveille, F},
title = {Staphylococcus aureus Strain-Dependent Biofilm Formation in Bone-Like Environment.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {714994},
pmid = {34557170},
issn = {1664-302X},
abstract = {Staphylococcus aureus species is an important threat for hospital healthcare because of frequent colonization of indwelling medical devices such as bone and joint prostheses through biofilm formations, leading to therapeutic failure. Furthermore, bacteria within biofilm are less sensitive to the host immune system responses and to potential antibiotic treatments. We suggested that the periprosthetic bone environment is stressful for bacteria, influencing biofilm development. To provide insights into S. aureus biofilm properties of three strains [including one methicillin-resistant S. aureus (MRSA)] under this specific environment, we assessed several parameters related to bone conditions and expected to affect biofilm characteristics. We reported that the three strains harbored different behaviors in response to the lack of oxygen, casamino acids and glucose starvation, and high concentration of magnesium. Each strain presented different biofilm biomass and live adherent cells proportion, or matrix production and composition. However, the three strains shared common responses in a bone-like environment: a similar production of extracellular DNA and engagement of the SOS response. This study is a step toward a better understanding of periprosthetic joint infections and highlights targets, which could be common among S. aureus strains and for future antibiofilm strategies.},
}

@article {pmid34556107,
year = {2021},
author = {Poornima, P and Krithikadatta, J and Ponraj, RR and Velmurugan, N and Kishen, A},
title = {Biofilm formation following chitosan-based varnish or chlorhexidine-fluoride varnish application in patients undergoing fixed orthodontic treatment: a double blinded randomised controlled trial.},
journal = {BMC oral health},
volume = {21},
number = {1},
pages = {465},
pmid = {34556107},
issn = {1472-6831},
abstract = {BACKGROUND: Orthodontic treatment poses an increased risk of plaque accumulation and demineralisation of enamel leading to white spot lesion around the brackets. This parallel arm trial aims to assess the degree of bacterial plaque formation adjacent to orthodontic brackets, following the application of a chitosan-based varnish or chlorhexidene-fluoride varnish.

METHODS: A total of 200 teeth from 20 patients undergoing fixed orthodontic therapy were assessed and biofilm formation around the brackets were recorded using the Bonded Bracket Index (Plaque index) at baseline and weekly for 6 weeks. The bacterial count and plaque pH at corresponding weekly intervals were also recorded. Following bracket bonding, the patients were cluster randomised to receive chitosan-based varnish-CHS (UNO Gel Bioschell, Germiphene corp., Brantford, Canada) or chlorhexidine-fluoride varnish-CFV (Cervitec F, Ivoclar Vivadent, Schaan, Liechtenstein) every week on the representative teeth respectively. BBI proportions were compared between groups at all time intervals using Chi square test. Mean plaque bacterial count and plaque pH were compared using Mann Whitney U test and Tukey's HSD test respectively.

RESULTS: Baseline characteristics were similar between the groups: Mean age was CHS = 23 and CFV = 21; male to female ratio was CHS = 5/5, CFV = 7/3. At the end of 6 weeks, chitosan-based varnish performed equal to chlorhexidine-fluoride varnish (P > 0.05) with 98% and 95% of teeth with acceptable scores respectively. The plaque bacterial count significantly reduced at 6 weeks for both varnish compared to the baseline; The value for CHS was 0.43 ± 0.4 × 104 and CFV was 0.77 ± 0.64 × 104 CFU (P < 0.05), with no difference between both the varnishes. Both varnishes had no effect on the plaque pH that remained neutral.

CONCLUSION: This trial showed that both chitosan-based varnish and chlorhexidine-fluoride varnish reduced bacterial count, while the plaque pH remained neutral over a period of six weeks in patients undergoing fixed orthodontic therapy. The anti-plaque effects of the natural biopolymeric chitosan-based varnish was similar to that of chlorhexidine-fluoride varnish, a known chemotherapeutic agent. Registration: This trial protocol was registered with https://www.ctri.nic.in (CTRI/2019/05/018896). (Date of registration 02/05/2019).

PROTOCOL: The protocol was not published before trial commencement.},
}

@article {pmid34554005,
year = {2021},
author = {Martinez, JN and Nishihara, A and Haruta, S},
title = {Metagenome-Assembled Genome Sequences Recovered from Epilithic River Biofilm in the Tama River, Japan.},
journal = {Microbiology resource announcements},
volume = {10},
number = {38},
pages = {e0066421},
doi = {10.1128/MRA.00664-21},
pmid = {34554005},
issn = {2576-098X},
support = {//Tokyu Foundation for a Better Environment/ ; },
abstract = {Draft genome sequences of putatively novel bacteria were assembled from the metagenome of epilithic biofilm samples collected from the Tama River (Tokyo, Japan). The metagenome contains 44,630,724 sequences, 44,792 contigs, and 48% G+C content. Binning resulted in 31 metagenome-assembled genomes (MAGs) with ≥50% completeness.},
}

@article {pmid34554002,
year = {2021},
author = {Rajeev, M and Sushmitha, TJ and Toleti, SR and Pandian, SK},
title = {Draft Genome Sequencing of Pseudoalteromonas tetraodonis Strain kknpp56, a Potent Biofilm-Forming Bacterium Isolated from Early-Stage Marine Biofilm.},
journal = {Microbiology resource announcements},
volume = {10},
number = {38},
pages = {e0060521},
doi = {10.1128/MRA.00605-21},
pmid = {34554002},
issn = {2576-098X},
support = {51/14/06/2019-BRNS//DAE | Board of Research in Nuclear Sciences (BRNS)/ ; 51/14/06/2019-BRNS//DAE | Board of Research in Nuclear Sciences (BRNS)/ ; 51/14/06/2019-BRNS//DAE | Board of Research in Nuclear Sciences (BRNS)/ ; 51/14/06/2019-BRNS//DAE | Board of Research in Nuclear Sciences (BRNS)/ ; },
abstract = {Pseudoalteromonas tetraodonis strain kknpp56 is an exopolysaccharide (EPS)-producing marine bacterium that forms potent biofilm. To determine the biosynthesis pathways involved in the EPS production of this bacterium, whole-genome sequencing was performed. The complete genome comes from one chromosome containing 3.72 Mbp of DNA with a G+C content of 41%.},
}

@article {pmid34553907,
year = {2021},
author = {Wang, J and Guo, X and Xue, J},
title = {Biofilm-Developed Microplastics As Vectors of Pollutants in Aquatic Environments.},
journal = {Environmental science & technology},
volume = {},
number = {},
pages = {},
doi = {10.1021/acs.est.1c04466},
pmid = {34553907},
issn = {1520-5851},
abstract = {Microplastics are a big and growing part of global pollution, which has aroused increasing concern in recent years because of their large amount, wide distribution, and adverse effects. Microplastics can sorb various pollutants from aquatic environments and act as vectors of pollutants. Most studies mainly focused on the virgin microplastics. However, microplastics in environments can be easily colonized by microorganisms, and form biofilm, which will influence the behaviors and potential risks of microplastics. The formation of biofilm on microplastics and its effects on their properties have been studied before, but their sorption and transport behaviors, and potential risks for pollutants' transfer have not been reviewed. In this paper, the role of biofilm-developed microplastics as vectors of pollutants was thoroughly analyzed and summarized. First, the formation of biofilm on microplastics, the compositions of microorganisms in biofilm, the influencing factors, and the property changes of microplastics after biofilm attachment are thoroughly reviewed. Second, the sorption of pollutants onto biofilm-developed microplastics is discussed. Third, the role of biofilm-developed microplastics as vector of pollutants are analyzed. We concluded that microplastics could provide unique substrates for microorganisms. Biofilm-developed microplastics can sorb more pollutants than the virgin ones, then act as vectors to introduce pollutants and attached microorganisms to aquatic environments and to organisms.},
}

@article {pmid34550756,
year = {2021},
author = {Stanford, K and Tran, F and Zhang, P and Yang, X},
title = {Biofilm-forming capacity of Escherichia coli isolated from cattle and beef packing plants: relation to virulence attributes, stage of processing, antimicrobial interventions, and heat tolerance.},
journal = {Applied and environmental microbiology},
volume = {},
number = {},
pages = {AEM0112621},
doi = {10.1128/AEM.01126-21},
pmid = {34550756},
issn = {1098-5336},
abstract = {Despite the importance of biofilm formation in contamination of meat by pathogenic Escherichia coli at slaughter plants, drivers for biofilm remain unclear. To identify selection pressures for biofilm, we evaluated 745 isolates from cattle and 700 generic E. coli from two beef slaughter plants for motility, expression of curli and cellulose, and biofilm-forming potential. Cattle isolates were also screened for serogroup, stx1, stx2, eae and rpoS. Generic E. coli were compared by source (hide of carcass, hide-off carcass, processing equipment) before and after implementation of antimicrobial hurdles. The proportion of E. coli capable of forming biofilms was lowest (7.1%; P < 0.05) for cattle isolates and highest (87.3%; P < 0.05) from equipment. Only one enterohemorrhagic E. coli (EHEC) was an extremely-strong biofilm-former, in contrast to 73.4% of E. coli from equipment. Isolates from equipment after sanitation had a greater biofilm-forming capacity (P < 0.001) than those before sanitation. Most cattle isolates were motile and expressed curli, although these traits along with expression of cellulose and detection of rpoS were not necessary for biofilm formation. In contrast, isolates capable of forming biofilms on equipment were almost exclusively motile and able to express curli. Results of the present study indicate that cattle would rarely carry EHEC capable of making strong biofilms to slaughter plants. However, if biofilm-forming EHEC contaminated equipment, current sanitation procedures may not eliminate the most robust biofilm-forming strains. Accordingly, new and effective anti-biofilm hurdles are required for meat-processing equipment to reduce future instances of food-borne disease. Importance As the majority of enterohemorrhagic E. coli (EHEC) are not capable of forming biofilms, sources were undetermined of the biofilm-forming EHEC isolated from 'high-event periods' in beef slaughter plants. This study demonstrated that sanitation procedures used on beef-processing equipment may inadvertently lead to survival of robust biofilm-forming strains of E. coli. Cattle only rarely carry EHEC capable of forming strong biofilms (1/745 isolates evaluated), but isolates with greater biofilm-forming capacity were more likely (P < 0.001) to survive equipment sanitation. In contrast, chilling carcasses for 3 days at 0°C reduced (P < 0.05) the proportion of biofilm-forming E. coli. Consequently, an additional anti-biofilm hurdle for meat-processing equipment, perhaps involving cold exposure, is necessary to further reduce the risk of food-borne disease.},
}

@article {pmid34550209,
year = {2021},
author = {Silva, AMCMD and Costa Júnior, SD and Lima, JLC and Farias Filho, JLB and Cavalcanti, IMF and Maciel, MAV},
title = {Investigation of the association of virulence genes and biofilm production with infection and bacterial colonization processes in multidrug-resistant Acinetobacter spp.},
journal = {Anais da Academia Brasileira de Ciencias},
volume = {93},
number = {suppl 3},
pages = {e20210245},
doi = {10.1590/0001-3765202120210245},
pmid = {34550209},
issn = {1678-2690},
mesh = {*Acinetobacter Infections ; *Acinetobacter baumannii/genetics ; Anti-Bacterial Agents/pharmacology ; Biofilms ; Drug Resistance, Multiple, Bacterial/genetics ; Humans ; Microbial Sensitivity Tests ; Virulence/genetics ; },
abstract = {The aim of this study was to evaluate the phenotypic and molecular patterns of biofilm formation in infection and colonization isolates of Acinetobacter spp. from patients who were admitted in a public hospital of Recife-PE-Brazil in 2018-2019. For the biofilm phenotypic analysis, Acinetobacter spp. isolates were evaluated by the crystal violet staining method; the search of virulence genes (bap, ompA, epsA, csuE and bfmS) was performed by PCR; and the ERIC-PCR was performed for molecular typing. Amongst the 38 Acinetobacter spp. isolates, 20 were isolated from infections and 18 from colonization. The resistance profile pointed that 86.85% (33/38) of the isolates were multidrug-resistant, being three infection isolates, and two colonization isolates resistant to polymyxin B. All the isolates were able to produce biofilm and they had at least one of the investigated virulence genes on their molecular profile, but the bap gene was found in 100% of them. No clones were detected by ERIC-PCR. There was no correlation between biofilm formation and the resistance profile of the bacteria, neither to the molecular profile of the virulence genes. Thus, the ability of Acinetobacter spp. to form biofilm is probably related to the high frequency of virulence genes.},
}

*Acinetobacter Infections
*Acinetobacter baumannii/genetics
Anti-Bacterial Agents/pharmacology
Biofilms
Drug Resistance, Multiple, Bacterial/genetics
Humans
Microbial Sensitivity Tests
Virulence/genetics

@article {pmid34550006,
year = {2021},
author = {Wu, Y and Meng, Y and Qian, L and Ding, B and Han, H and Chen, H and Bai, L and Qu, D and Wu, Y},
title = {The Vancomycin Resistance-Associated Regulatory System VraSR Modulates Biofilm Formation of Staphylococcus epidermidis in an ica-Dependent Manner.},
journal = {mSphere},
volume = {},
number = {},
pages = {e0064121},
doi = {10.1128/mSphere.00641-21},
pmid = {34550006},
issn = {2379-5042},
abstract = {The two-component system VraSR responds to the cell wall-active antibiotic stress in Staphylococcus epidermidis. To study its regulatory function in biofilm formation, a vraSR deletion mutant (ΔvraSR) was constructed using S. epidermidis strain 1457 (SE1457) as the parent strain. Compared to SE1457, the ΔvraSR mutant showed impaired biofilm formation both in vitro and in vivo with a higher ratio of dead cells within the biofilm. Consistently, the ΔvraSR mutant produced much less polysaccharide intercellular adhesin (PIA). The ΔvraSR mutant also showed increased susceptibility to the cell wall inhibitor and SDS, and its cell wall observed under a transmission electron microscope (TEM) appeared to be thinner and interrupted, which is in accordance with higher susceptibility to the stress. Complementation of vraSR in the ΔvraSR mutant restored the biofilm formation and the cell wall thickness to wild-type levels. Transcriptome sequencing (RNA-Seq) showed that the vraSR deletion affected the transcription levels of 73 genes, including genes involved in biofilm formation, bacterial programmed cell death (CidA-LrgAB system), glycolysis/gluconeogenesis, the pentose phosphate pathway (PPP), and the tricarboxylic acid (TCA) cycle, etc. The results of RNA-Seq were confirmed by quantitative real-time reverse transcription-PCR (qRT-PCR). In the ΔvraSR mutant, the expression of icaA and lrgAB was downregulated and the expression of icaR and cidA was upregulated, in comparison to that of SE1457. The transcriptional levels of antibiotic-resistant genes (pbp2, serp1412, murAA, etc.) had no significant changes. An electrophoretic mobility shift assay further revealed that phosphorylated VraR bound to the promoter regions of the ica operon, as well as its own promoter region. This study demonstrates that in S. epidermidis, VraSR is an autoregulator and directly regulates biofilm formation in an ica-dependent manner. Upon cell wall stress, it indirectly regulates cell death and drug resistance in association with alterations to multiple metabolism pathways. IMPORTANCE S. epidermidis is a leading cause of hospital-acquired catheter-related infections, and its pathogenicity depends mostly on its ability to form biofilms on implants. The biofilm formation is a complex procedure that involves multiple regulating factors. Here, we show that a vancomycin resistance-associated two-component regulatory system, VraSR, plays an important role in modulating S. epidermidis biofilm formation and tolerance to stress. We demonstrate that S. epidermidis VraSR is an autoregulated system that selectively responds to stress targeting cell wall synthesis. Besides, phosphorylated VraR can bind to the promoter region of the ica operon and directly regulates polysaccharide intercellular adhesin production and biofilm formation in S. epidermidis. Furthermore, VraSR may indirectly modulate bacterial cell death and extracellular DNA (eDNA) release in biofilms through the CidA-LrgAB system. This work provides a new molecular insight into the mechanisms of VraSR-mediated modulation of the biofilm formation and cell death of S. epidermidis.},
}

@article {pmid34548785,
year = {2021},
author = {Amankwah, S and Abdella, K and Kassa, T},
title = {Bacterial Biofilm Destruction: A Focused Review On The Recent Use of Phage-Based Strategies With Other Antibiofilm Agents.},
journal = {Nanotechnology, science and applications},
volume = {14},
number = {},
pages = {161-177},
pmid = {34548785},
issn = {1177-8903},
abstract = {Biofilms are bacterial communities that live in association with biotic or abiotic surfaces and enclosed in an extracellular polymeric substance. Their formation on both biotic and abiotic surfaces, including human tissue and medical device surfaces, pose a major threat causing chronic infections. In addition, current antibiotics and antiseptic agents have shown limited ability to completely remove biofilms. In this review, the authors provide an overview on the formation of bacterial biofilms and its characteristics, burden and evolution with phages. Moreover, the most recent possible use of phages and phage-derived enzymes to combat bacteria in biofilm structures is elucidated. From the emerging results, it can be concluded that despite successful use of phages and phage-derived products in destroying biofilms, they are mostly not adequate to eradicate all bacterial cells. Nevertheless, a combined therapy with the use of phages and/or phage-derived products with other antimicrobial agents including antibiotics, nanoparticles, and antimicrobial peptides may be effective approaches to remove biofilms from medical device surfaces and to treat their associated infections in humans.},
}

@article {pmid34546549,
year = {2021},
author = {Mukhi, M and Vishwanathan, AS},
title = {Identifying potential inhibitors of biofilm-antagonistic proteins to promote biofilm formation: a virtual screening and molecular dynamics simulations approach.},
journal = {Molecular diversity},
volume = {},
number = {},
pages = {},
pmid = {34546549},
issn = {1573-501X},
abstract = {Microbial biofilms play a critical role in environmental biotechnology and associated applications. Biofilm production can be enhanced by inhibiting the function of proteins that negatively regulate their formation. With this objective, an in silico approach was adopted to identify competitive inhibitors of eight biofilm-antagonistic proteins, namely AbrB and SinR (from Bacillus subtilis) and AmrZ, PDE (EAL), PslG, RetS, ShrA and TpbA (from Pseudomonas aeruginosa). Fifteen inhibitors that structurally resembled the natural ligand of each protein were shortlisted using ligand-based and structure-based virtual screening. The top four inhibitors obtained from molecular docking using Autodock Vina were further docked using SwissDock and DOCK 6.9 to obtain a consensus hit for each protein based on different scoring functions. Further analysis of the protein-ligand complexes revealed that these top inhibitors formed significant non-covalent interactions with their respective protein binding sites. The eight protein-ligand complexes were then subjected to molecular dynamics simulations for 30 ns using GROMACS. RMSD and radius of gyration values of 0.1-0.4 nm and 1.0-3.5 nm, respectively, along with hydrogen bond formation throughout the trajectory indicated that all the complexes remained stable, compact and intact during the simulation period. Binding energy values between -20 and -77 kJ/mol obtained from MM-PBSA calculations further confirmed the high affinities of the eight inhibitors for their respective receptors. The outcome of this study holds great promise to enhance biofilms that are central to biotechnological processes associated with microbial electrochemical technologies, wastewater treatment, bioremediation and the industrial production of value-added products.},
}

@article {pmid34543761,
year = {2021},
author = {Ali Mohammed, MM and Pettersen, VK and Nerland, AH and Wiker, HG and Bakken, V},
title = {Label-free quantitative proteomic analysis of the oral bacteria Fusobacterium nucleatum and Porphyromonas gingivalis to identify protein features relevant in biofilm formation.},
journal = {Anaerobe},
volume = {72},
number = {},
pages = {102449},
doi = {10.1016/j.anaerobe.2021.102449},
pmid = {34543761},
issn = {1095-8274},
abstract = {BACKGROUND: The opportunistic pathogens Fusobacterium nucleatum and Porphyromonas gingivalis are Gram-negative bacteria associated with oral biofilm and periodontal disease. This study investigated interactions between F. nucleatum and P. gingivalis proteomes with the objective to identify proteins relevant in biofilm formation.

METHODS: We applied liquid chromatography-tandem mass spectrometry to determine the expressed proteome of F. nucleatum and P. gingivalis cells grown in biofilm or planktonic culture, and as mono- and dual-species models. The detected proteins were classified into functional categories and their label-free quantitative (LFQ) intensities statistically compared.

RESULTS: The proteomic analyses detected 1,322 F. nucleatum and 966 P. gingivalis proteins, including abundant virulence factors. Using univariate statistics, we identified significant changes between biofilm and planktonic culture (p-value ≤0.05) in 0,4% F. nucleatum, 7% P. gingivalis, and 14% of all proteins in the dual-species model. For both species, proteins involved in vitamin B2 (riboflavin) metabolism had significantly increased levels in biofilm. In both mono- and dual-species biofilms, P. gingivalis increased the production of proteins for translation, oxidation-reduction, and amino acid metabolism compared to planktonic cultures. However, when we compared LFQ intensities between mono- and dual-species, over 90% of the significantly changed P. gingivalis proteins had their levels reduced in biofilm and planktonic settings of the dual-species model.

CONCLUSIONS: The findings suggest that P. gingivalis reduces the production of multiple proteins because of the F. nucleatum presence. The results highlight the complex interactions of bacteria contributing to oral biofilms, which need to be considered in the design of prevention strategies.},
}

@article {pmid34543106,
year = {2021},
author = {Thai, SN and Lum, MR and Naegle, J and Onofre, M and Abdulla, H and Garcia, A and Fiterz, A and Arnell, A and Lwin, TT and Kavanaugh, A and Hikmat, Z and Garabedian, N and Ngo, RT and Dimaya, B and Escamilla, A and Barseghyan, L and Shibatsuji, M and Soltani, S and Butcher, L and Hikmat, F and Amirian, D and Bazikyan, A and Brandt, N and Sarkisian, M and Munoz, X and Ovakimyan, A and Burnett, E and Pham, JN and Shirvanian, A and Hernandez, R and Vardapetyan, M and Wada, M and Ramirez, C and Zakarian, M and Billi, F},
title = {Multiple copies of flhDC in Paraburkholderia unamae regulate flagellar gene expression, motility, and biofilm formation.},
journal = {Journal of bacteriology},
volume = {},
number = {},
pages = {JB0029321},
doi = {10.1128/JB.00293-21},
pmid = {34543106},
issn = {1098-5530},
abstract = {FlhDC is a heterohexameric complex that acts as a master regulator of flagellar biosynthesis genes in numerous bacteria. Previous studies have identified a single flhDC operon encoding this complex. However, we found that two flhDC loci are present throughout Paraburkholderia and two additional flhC copies also present in P. unamae. Systematic deletion analysis in P. unamae of the different flhDC copies showed that one of the operons, flhDC1, plays the predominant role, with deletion of its genes resulting in a severe inhibition of motility and biofilm formation. Expression analysis using promoter-lacZ fusions and real-time quantitative PCR support the primary role of flhDC1 in flagellar gene regulation, with flhDC2 a secondary contributor. Phylogenetic analysis shows the presence of the flhDC1 and flhDC2 operons throughout Paraburkholderia. In contrast, Burkholderia and other bacteria only carry the copy syntenous with flhDC2. The varying impact each copy of flhDC has on downstream processes indicates that regulation of FlhDC in P. unamae, and likely other Paraburkholderia, is regulated at least in part by the presence of multiple copies of these genes. IMPORTANCE Motility is important in the colonization of plant roots by beneficial and pathogenic bacteria, with flagella playing essential roles in host cell adhesion, entrance, and biofilm formation. Flagellar biosynthesis is energetically expensive. Its complex regulation by the FlhDC master regulator is well-studied in peritrichous flagella expressing enterics. We report the unique presence throughout Paraburkholderia of multiple copies of flhDC. In P. unamae, the flhDC1 copy showed higher expression and greater effect on swim motility, flagellar development, and regulation of downstream genes, than the flhDC2 copy that is syntenous to flhDC in E. coli and pathogenic Burkholderia spp. The flhDC genes have evolved differently in these plant-growth promoting bacteria, giving an additional layer of complexity in gene regulation by FlhDC.},
}

@article {pmid34540932,
year = {2021},
author = {Santos, LM and Rodrigues, DM and Kalil, MA and Azevedo, V and Meyer, R and Umsza-Guez, MA and Machado, BA and Seyffert, N and Portela, RW},
title = {Activity of Ethanolic and Supercritical Propolis Extracts in Corynebacterium pseudotuberculosis and Its Associated Biofilm.},
journal = {Frontiers in veterinary science},
volume = {8},
number = {},
pages = {700030},
pmid = {34540932},
issn = {2297-1769},
abstract = {Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis in small ruminants, a chronic disease characterized by the development of granulomas in superficial and visceral lymph nodes as well as in several organs. An important characteristic of the infection with this bacterium is the formation of a biofilm and the absence of effective antibiotic therapy against the disease. From this scenario, the objective of this study was to evaluate the susceptibility of C. pseudotuberculosis to conventional antibiotics and to red, green, and brown propolis extracts obtained by the supercritical and ethanolic extraction methods as well as its activity in the bacterial biofilm. The results of the sensitivity test using antibiotics indicated a sensitivity of C. pseudotuberculosis strains to the antimicrobial agents. The ethanolic extract of green propolis and the supercritical red propolis extract showed the best antibacterial activities against planktonic C. pseudotuberculosis. A lower antimicrobial activity of the brown propolis extract was identified. Propolis extracts were effective in interfering with the formation of the C. pseudotuberculosis biofilm but had little activity on the consolidated biofilm. In conclusion, propolis extracts are more effective against C. pseudotuberculosis in the planktonic stage, being able to interfere with the formation of bacterial biofilm. However, the action of propolis extracts in a sessile and structured microbial biofilm is reduced.},
}

@article {pmid34539591,
year = {2021},
author = {Eckroat, TJ and Greguske, C and Hunnicutt, DW},
title = {The Type 9 Secretion System Is Required for Flavobacterium johnsoniae Biofilm Formation.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {660887},
pmid = {34539591},
issn = {1664-302X},
abstract = {Flavobacterium johnsoniae forms biofilms in low nutrient conditions. Protein secretion and cell motility may have roles in biofilm formation. The F. johnsoniae type IX secretion system (T9SS) is important for both secretion and motility. To determine the roles of each process in biofilm formation, mutants defective in secretion, in motility, or in both processes were tested for their effects on biofilm production using a crystal violet microplate assay. All mutants that lacked both motility and T9SS-mediated secretion failed to produce biofilms. A porV deletion mutant, which was severely defective for secretion, but was competent for motility, also produced negligible biofilm. In contrast, mutants that retained secretion but had defects in gliding formed biofilms. An sprB mutant that is severely but incompletely defective in gliding motility but retains a fully functional T9SS was similar to the wild type in biofilm formation. Mutants with truncations of the gldJ gene that compromise motility but not secretion showed partial reduction in biofilm formation compared to wild type. Unlike the sprB mutant, these gldJ truncation mutants were essentially nonmotile. The results show that a functional T9SS is required for biofilm formation. Gliding motility, while not required for biofilm formation, also appears to contribute to formation of a robust biofilm.},
}

@article {pmid34537868,
year = {2021},
author = {Ishvaria, S and Dharshini, RS and Manickam, R and Pooja, KR and Ramya, M},
title = {Draft genome sequencing and functional annotation and characterization of biofilm-producing bacterium Bacillus novalis PD1 isolated from rhizospheric soil.},
journal = {Antonie van Leeuwenhoek},
volume = {},
number = {},
pages = {},
pmid = {34537868},
issn = {1572-9699},
abstract = {Biofilm forming bacterium Bacillus novalis PD1 was isolated from the rhizospheric soil of a paddy field. B. novalis PD1 is a Gram-positive, facultatively anaerobic, motile, slightly curved, round-ended, and spore-forming bacteria. The isolate B. novalis PD1 shares 98.45% similarity with B. novalis KB27B. B. vireti LMG21834 and B. drentensis NBRC 102,427 are the closest phylogenetic neighbours for B. novalis PD1. The draft genome RAST annotation showed a linear chromosome with 4,569,088 bp, encoding 6139 coding sequences, 70 transfer RNA (tRNA), and 11 ribosomal RNA (rRNA) genes. The genomic annotation of biofilm forming B. novalis PD1(> 3.6@OD595nm) showed the presence of exopolysaccharide-forming genes (ALG, PSL, and PEL) as well as other biofilm-related genes (comER, Spo0A, codY, sinR, TasA, sipW, degS, and degU). Antibiotic inactivation gene clusters (ANT (6)-I, APH (3')-I, CatA15/A16 family), efflux pumps conferring antibiotic resistance genes (BceA, BceB, MdtABC-OMF, MdtABC-TolC, and MexCD-OprJ), and secondary metabolites linked to phenazine, terpene, and beta lactone gene clusters are part of the genome.},
}

@article {pmid34534802,
year = {2021},
author = {Miao, L and Guo, S and Wu, J and Adyel, TM and Liu, Z and Liu, S and Hou, J},
title = {Polystyrene nanoplastics change the functional traits of biofilm communities in freshwater environment revealed by GeoChip 5.0.},
journal = {Journal of hazardous materials},
volume = {423},
number = {Pt B},
pages = {127117},
doi = {10.1016/j.jhazmat.2021.127117},
pmid = {34534802},
issn = {1873-3336},
abstract = {There is an increasing concern regarding the potential effects of nanoplastics (NPs) on freshwater ecosystems. Considering the functional values of biofilms in freshwater, knowledge on whether and to what extent NPs can influence the ecosystem processes of biofilms were still limited. Herein, the freshwater biofilms cultured in lab were exposed to 100 nm polystyrene NPs (PS-NPs) of different dosages (1 and 10 mg/L) for 14 days. Confocal laser scanning microscope observation indicated that biofilms were dominated by filamentous, and spiral algae species and the intensity of extracellular polymeric substances increased under PS-NPs exposure. GeoChip 5.0 analysis revealed that PS-NPs exposure triggered a significant increase in functional genes α diversity (p < 0.05) and altered biofilms' functional structure. Furthermore, the abundance of genes involved in the total carbon and nitrogen cycling were increased under PS-NPs exposure. The abundance of nitrogen fixation genes experienced the most pronounced increase (24.4%) under 1 mg/L PS-NPs treatment, consistent with the increase of ammonium in overlying water. Whereas antibiotic resistance genes and those related to photosynthetic pigments production were suppressed. These results provided direct evidence for PS-NPs' effects on the biofilm functions in terms of biogeochemical cycling, improving our understanding of the potentials of NPs on freshwater ecosystems.},
}

@article {pmid34531666,
year = {2021},
author = {Gedefie, A and Demsis, W and Ashagrie, M and Kassa, Y and Tesfaye, M and Tilahun, M and Bisetegn, H and Sahle, Z},
title = {Acinetobacter baumannii Biofilm Formation and Its Role in Disease Pathogenesis: A Review.},
journal = {Infection and drug resistance},
volume = {14},
number = {},
pages = {3711-3719},
pmid = {34531666},
issn = {1178-6973},
abstract = {Acinetobacter species, particularly Acinetobacter baumannii, is the first pathogen on the critical priority list of pathogens for novel antibiotics to become a "red-alert" human pathogen. Acinetobacter baumannii is an emerging global antibiotic-resistant gram-negative bacteria that most typically causes biofilm-associated infections such as ventilator-associated pneumonia and catheter-related infection, both of which are resistant to antibiotic therapy. A. baumannii's capacity to develop antibiotic resistance mechanisms allows the organism to thrive in hospital settings, facilitating the global spread of multidrug-resistant strains. Although Acinetobacter infections are quickly expanding throughout hospital environments around the world, the highest concentration of infections occurs in intensive care units (ICUs). Biofilms are populations of bacteria on biotic or abiotic surfaces that are encased in the extracellular matrix and play a crucial role in pathogenesis, making treatment options more difficult. Even though a variety of biological and environmental elements are involved in the production of A. baumannii biofilms, glucose is the most important component. Biofilm-mediated A. baumannii infections are the most common type of A. baumannii infection associated with medical equipment, and they are extremely difficult to treat. As a result, health care workers (HCWs) should focus on infection prevention and safety actions to avoid A. baumannii biofilm-related infections caused by medical devices, and they should be very selective when using treatments in combination with anti-biofilms. Therefore, this review discusses biofilm formation in A. baumannii, its role in disease pathogenesis, and its antimicrobial resistance mechanism.},
}

@article {pmid34530240,
year = {2021},
author = {Liu, C and Yan, H and Sun, Y and Chen, B},
title = {Contribution of enrofloxacin and Cu2+ to the antibiotic resistance of bacterial community in a river biofilm.},
journal = {Environmental pollution (Barking, Essex : 1987)},
volume = {291},
number = {},
pages = {118156},
doi = {10.1016/j.envpol.2021.118156},
pmid = {34530240},
issn = {1873-6424},
abstract = {Pollutants discharged from wastewater are the main cause of the spread of antibiotic resistance in river biofilms. There is controversy regarding the primary contribution of environmental selectors such as antibiotics and heavy metals to the development of antibiotic resistance in bacterial communities. Here, this study compared the effect of environmental safety concentration Cu2+ and enrofloxacin (ENR) on the evolution of antibiotic resistance by examining phenotypic characteristics and genotypic profiles of bacterial communities in a river biofilm, and then distinguished the major determinants from a comprehensive perspective. The pollution induced community tolerance in ENR-treated group was significantly higher than that in Cu2+-treated group (at concentration levels of 100 and 1000 μg/L). Metagenomic sequencing results showed that ENR significantly increased the number and total abundance of antibiotic resistance genes (ARGs), but there was no significant change in the Cu2+- treated group. Compared with Cu2+, ENR was the major selective agent in driving the change of taxonomic composition because the taxonomic composition in ENR was the most different from the original biofilm. Comparing and analyzing the prokaryotic composition, the phylum of Proteobacteria was enriched in both ENR and Cu2+ treated groups. Among them, Acidovorax and Bosea showed resistance to both pollutants. Linking taxonomic composition to ARGs revealed that the main potential hosts of fluoroquinolone resistance genes were Comamonas, Sphingopyxis, Bradyrhizobium, Afipia, Rhodopseudomonas, Luteimonas and Hoeflea. The co-occurrence of ARGs and metal resistance genes (MRGs) showed that the multidrug efflux pump was the key mechanism connecting MRGs and ARGs. Network analysis also revealed that the reason of Cu2+ selected for fluoroquinolones resistant bacterial communities was the coexistence of multidrug efflux gene and MRGs. Our research emphasizes the importance of antibiotics in promoting the development of antibiotic resistant bacterial communities from the perspective of changes in community structure and resistome in river biofilms.},
}

@article {pmid34530014,
year = {2021},
author = {Kasza, K and Gurnani, P and Hardie, KR and Cámara, M and Alexander, C},
title = {Challenges and solutions in polymer drug delivery for bacterial biofilm treatment: A tissue-by-tissue account.},
journal = {Advanced drug delivery reviews},
volume = {178},
number = {},
pages = {113973},
doi = {10.1016/j.addr.2021.113973},
pmid = {34530014},
issn = {1872-8294},
abstract = {To tackle the emerging antibiotic resistance crisis, novel antimicrobial approaches are urgently needed. Bacterial communities (biofilms) are a particular concern in this context. Biofilms are responsible for most human infections and are inherently less susceptible to antibiotic treatments. Biofilms have been linked with several challenging chronic diseases, including implant-associated osteomyelitis and chronic wounds. The specific local environments present in the infected tissues further contribute to the rise in antibiotic resistance by limiting the efficacy of systemic antibiotic therapies and reducing drug concentrations at the infection site, which can lead to reoccurring infections. To overcome the shortcomings of systemic drug delivery, encapsulation within polymeric carriers has been shown to enhance antimicrobial efficacy, permeation and retention at the infection site. In this Review, we present an overview of current strategies for antimicrobial encapsulation within polymeric carriers, comparing challenges and solutions on a tissue-by-tissue basis. We compare challenges and proposed drug delivery solutions from the perspective of the local environments for biofilms found in oral, wound, gastric, urinary tract, bone, pulmonary, vaginal, ocular and middle/inner ear tissues. We will also discuss future challenges and barriers to clinical translation for these therapeutics. The following Review demonstrates there is a significant imbalance between the research focus being placed on different tissue types, with some targets (oral and wound biofims) being extensively more studied than others (vaginal and otitis media biofilms and endocarditis). Furthermore, the importance of the local tissue environment when selecting target therapies is demonstrated, with some materials being optimal choices for certain sites of bacterial infection, while having limited applicability in others.},
}

@article {pmid34529974,
year = {2021},
author = {Ryan Kaler, KM and Nix, JC and Schubot, FD},
title = {RetS inhibits Pseudomonas aeruginosa biofilm formation by disrupting the canonical histidine kinase dimerization interface of GacS.},
journal = {The Journal of biological chemistry},
volume = {},
number = {},
pages = {101193},
doi = {10.1016/j.jbc.2021.101193},
pmid = {34529974},
issn = {1083-351X},
abstract = {Bacterial signaling histidine kinases (HKs) have long been postulated to function exclusively through linear signal transduction chains. However, several HKs have recently been shown to form complex multikinase networks (MKNs). The most prominent MKN, involving the enzymes RetS and GacS, controls the switch between the motile and biofilm lifestyles in the pathogenic bacterium Pseudomonas aeruginosa. While GacS promotes biofilm formation, RetS counteracts GacS using three distinct mechanisms. Two are dephosphorylating mechanisms. The third, a direct binding between the RetS and GacS HK regions, blocks GacS autophosphorylation. Focusing on the third mechanism, we determined the crystal structure of a co-complex between the HK region of RetS and the dimerization and histidine phosphotransfer (DHp) domain of GacS. This is the first reported structure of a complex between two distinct bacterial signaling HKs. In the complex the canonical HK homodimerization interface is replaced by a strikingly similar heterodimeric interface between RetS and GacS. We further demonstrate that GacS autophosphorylates in trans, thus explaining why the formation of a RetS-GacS complex inhibits GacS autophosphorylation. Using mutational analysis in conjunction with bacterial two-hybrid and biofilm assays we not only corroborate the biological role of the observed RetS-GacS interactions, but also identify a residue critical for the equilibrium between the RetS-GacS complex and the respective RetS and GacS homodimers. Collectively, our findings suggest that RetS and GacS form a domain-swapped hetero-oligomer during the planktonic growth phase of P. aeruginosa before unknown signals cause its dissociation and a relief of GacS inhibition to promote biofilm formation.},
}

@article {pmid34529734,
year = {2021},
author = {Yu, Y and Kim, YH and Cho, WH and Son, BS and Yeo, HJ},
title = {Biofilm microbiome in extracorporeal membrane oxygenator catheters.},
journal = {PloS one},
volume = {16},
number = {9},
pages = {e0257449},
pmid = {34529734},
issn = {1932-6203},
abstract = {Despite the formation of biofilms on catheters for extracorporeal membrane oxygenation (ECMO), some patients do not show bacteremia. To elucidate the specific linkage between biofilms and bacteremia in patients with ECMO, an improved understanding of the microbial community within catheter biofilms is necessary. Hence, we aimed to evaluate the biofilm microbiome of ECMO catheters from adults with (n = 6) and without (n = 15) bacteremia. The microbiomes of the catheter biofilms were evaluated by profiling the V3 and V4 regions of bacterial 16s rRNA genes using the Illumina MiSeq sequencing platform. In total, 2,548,172 reads, with an average of 121,341 reads per sample, were generated. Although alpha diversity was slightly higher in the non-bacteremic group, the difference was not statistically significant. In addition, there was no difference in beta diversity between the two groups. We found 367 different genera, of which 8 were present in all samples regardless of group; Limnohabitans, Flavobacterium, Delftia, Massilia, Bacillus, Candidatus, Xiphinematobacter, and CL0-1 showed an abundance of more than 1% in the sample. In particular, Arthrobacter, SMB53, Neisseria, Ortrobactrum, Candidatus Rhabdochlamydia, Deefgae, Dyella, Paracoccus, and Pedobacter were highly abundant in the bacteremic group. Network analysis indicated that the microbiome of the bacteremic group was more complex than that of the non-bacteremic group. Flavobacterium and CL0.1, which were abundant in the bacteremic group, were considered important genera because they connected different subnetworks. Biofilm characteristics in ECMO catheters varied according to the presence or absence of bacteremia. There were no significant differences in diversity between the two groups, but there were significant differences in the community composition of the biofilms. The biofilm-associated community was dynamic, with the bacteremic group showing very complex network connections within the microbiome.},
}

@article {pmid34529329,
year = {2021},
author = {Moradi, M and Fazlyab, M and Pourhajibagher, M and Chiniforush, N},
title = {Antimicrobial action of photodynamic therapy on Enterococcus faecalis biofilm using curing light, curcumin and riboflavin.},
journal = {Australian endodontic journal : the journal of the Australian Society of Endodontology Inc},
volume = {},
number = {},
pages = {},
doi = {10.1111/aej.12565},
pmid = {34529329},
issn = {1747-4477},
abstract = {The aim of this study was to assess the effect of antimicrobial photodynamic therapy (aPDT) with curcumin and riboflavin on three-week Enterococcus faecalis biofilm. At first the 15-mm root canals of 65 single rooted extracted human teeth (including maxillary incisors, mandibular and maxillary canines and mandibular premolars) were separated from the crown and were prepared with ProTaper instruments. After autoclave sterilisation, samples were inoculated with E. faecalis suspension, and incubated for three weeks. After ensuring biofilm formation by scanning electron microscopy (SEM) in two teeth, the remaining 63 teeth were randomly divided into seven groups (n = 9): aPDT + curcumin, aPDT + riboflavin, LED, curcumin, riboflavin, 5.25% NaOCl (positive control) and no intervention (negative control). For light source a LED unit with 390-480 nm wavelength (peak of 460 nm), power density of 1000 ± 100 mW cm-2 and mean energy density of 60 J cm-2 was used. The roots were horizontally sectioned into coronal, middle and apical thirds each with 5 mm thicknesses. Dentin chips with equal weight (1 ± 0.005 g) were collected from the root canal walls with Gates Glidden drills and were transferred into microtubes containing 1 mL of sterile saline and vortexed for 30 s. Next, 10 µL of the contents of each tube was serially diluted and eventually, 10 µL of each solution was cultured on BHI agar. The number of colony-forming units was determined. Data were analysed using the Kruskal-Wallis and Friedman tests. The colony reduction was not significantly different between NaOCl and either riboflavin + LED or Curcumin + LED. The 5.25% NaOCl group showed maximum reduction in colony count, compared with the negative control (P = 0.00). Groups with aPDT with Curcumin + LED (P = 0.005), and with riboflavin + LED (P = 0.011) showed significant reduction in colony count in all three canal thirds (P < 0.05) without any difference with one another. With significant reduction of E. faecalis colony count, aPDT with Curcumin and riboflavin can serve as an adjunct to routine root canal disinfection method.},
}

@article {pmid34528354,
year = {2021},
author = {Urvoy, M and Lami, R and Dreanno, C and Daudé, D and Rodrigues, AMS and Gourmelon, M and L'Helguen, S and Labry, C},
title = {Quorum sensing disruption regulates hydrolytic enzyme and biofilm production in estuarine bacteria.},
journal = {Environmental microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1111/1462-2920.15775},
pmid = {34528354},
issn = {1462-2920},
abstract = {Biofilms of heterotrophic bacteria cover organic matter aggregates and constitute hotspots of mineralization, primarily acting through extracellular hydrolytic enzyme production. Nevertheless, regulation of both biofilm and hydrolytic enzyme synthesis remains poorly investigated, especially in estuarine ecosystems. In this study, various bioassays, mass spectrometry and genomics approaches were combined to test the possible involvement of quorum sensing (QS) in these mechanisms. QS is a bacterial cell-cell communication system that relies notably on the emission of N-acylhomoserine lactones (AHLs). In our estuarine bacterial collection, we found that 28 strains (9%), mainly Vibrio, Pseudomonas and Acinetobacter isolates, produced at least 14 different types of AHLs encoded by various luxI genes. We then inhibited the AHL QS circuits of those 28 strains using a broad-spectrum lactonase preparation and tested whether biofilm production as well as β-glucosidase and leucine-aminopeptidase activities were impacted. Interestingly, we recorded contrasted responses, as biofilm production, dissolved and cell-bound β-glucosidase and leucine-aminopeptidase activities significantly increased in 4%-68% of strains but decreased in 0%-21% of strains. These findings highlight the key role of AHL-based QS in estuarine bacterial physiology and ultimately on biogeochemical cycles. They also point out the complexity of QS regulations within natural microbial assemblages. This article is protected by copyright. All rights reserved.},
}

@article {pmid34526981,
year = {2021},
author = {Abdul Hamid, AI and Cara, A and Diot, A and Laurent, F and Josse, J and Gueirard, P},
title = {Differential Early in vivo Dynamics and Functionality of Recruited Polymorphonuclear Neutrophils After Infection by Planktonic or Biofilm Staphylococcus aureus.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {728429},
pmid = {34526981},
issn = {1664-302X},
abstract = {Staphylococcus aureus is a human pathogen known for its capacity to shift between the planktonic and biofilm lifestyles. In vivo, the antimicrobial immune response is characterized by the recruitment of inflammatory phagocytes, namely polymorphonuclear neutrophils (PMNs) and monocytes/macrophages. Immune responses to planktonic bacteria have been extensively studied, but many questions remain about how biofilms can modulate inflammatory responses and cause recurrent infections in live vertebrates. Thus, the use of biologically sound experimental models is essential to study the specific immune signatures elicited by biofilms. Here, a mouse ear pinna model of infection was used to compare early innate immune responses toward S. aureus planktonic or biofilm bacteria. Flow cytometry and cytokine assays were carried out to study the inflammatory responses in infected tissues. These data were complemented with intravital confocal imaging analyses, allowing the real-time observation of the dynamic interactions between EGFP + phagocytes and bacteria in the ear pinna tissue of LysM-EGFP transgenic mice. Both bacterial forms induced an early and considerable recruitment of phagocytes in the ear tissue, associated with a predominantly pro-inflammatory cytokine profile. The inflammatory response was mostly composed of PMNs in the skin and the auricular lymph node. However, the kinetics of PMN recruitment were different between the 2 forms in the first 2 days post-infection (pi). Two hours pi, biofilm inocula recruited more PMNs than planktonic bacteria, but with decreased motility parameters and capacity to emit pseudopods. Inversely, biofilm inocula recruited less PMNs 2 days pi, but with an "over-activated" status, illustrated by an increased phagocytic activity, CD11b level of expression and ROS production. Thus, the mouse ear pinna model allowed us to reveal specific differences in the dynamics of recruitment and functional properties of phagocytes against biofilms. These differences would influence the specific adaptive immune responses to biofilms elicited in the lymphoid tissues.},
}

@article {pmid34526980,
year = {2021},
author = {Prados, MB and Lescano, M and Porzionato, N and Curutchet, G},
title = {Wiring Up Along Electrodes for Biofilm Formation.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {726251},
pmid = {34526980},
issn = {1664-302X},
abstract = {Millimeter-length cables of bacteria were discovered growing along a graphite-rod electrode serving as an anode of a microbial electrolysis cell (MEC). The MEC had been inoculated with a culture of Fe-reducing microorganisms enriched from a polluted river sediment (Reconquista river, Argentina) and was operated at laboratory controlled conditions for 18 days at an anode poised potential of 240 mV (vs. Ag/AgCl), followed by 23 days at 480 mV (vs. Ag/AgCl). Anode samples were collected for scanning electron microscopy, phylogenetic and electrochemical analyses. The cables were composed of a succession of bacteria covered by a membranous sheath and were distinct from the known "cable-bacteria" (family Desulfobulbaceae). Apparently, the formation of the cables began with the interaction of the cells via nanotubes mostly located at the cell poles. The cables seemed to be further widened by the fusion between them. 16S rRNA gene sequence analysis confirmed the presence of a microbial community composed of six genera, including Shewanella, a well-characterized electrogenic bacteria. The formation of the cables might be a way of colonizing a polarized surface, as determined by the observation of electrodes extracted at different times of MEC operation. Since the cables of bacteria were distinct from any previously described, the results suggest that bacteria capable of forming cables are more diverse in nature than already thought. This diversity might render different electrical properties that could be exploited for various applications.},
}

@article {pmid34525984,
year = {2021},
author = {Schneider-Rayman, M and Steinberg, D and Sionov, RV and Friedman, M and Shalish, M},
title = {Effect of epigallocatechin gallate on dental biofilm of Streptococcus mutans: An in vitro study.},
journal = {BMC oral health},
volume = {21},
number = {1},
pages = {447},
pmid = {34525984},
issn = {1472-6831},
abstract = {BACKGROUND: Streptococcus mutans (S. mutans) plays a major role in the formation of dental caries. The aim of this study was to examine the effect of the green tea polyphenol, epigallocatechin gallate (EGCG), on biofilm formation of S. mutans.

METHODS: Following exposure to increasing concentrations of EGCG, the planktonic growth was measured by optical density and the biofilm biomass was quantified by crystal violet staining. Exopolysaccharides (EPS) production was visualized by confocal scanning laser microscopy, and the bacterial DNA content was determined by quantitative polymerase chain reaction (qPCR). Gene expression of selected genes was analyzed by real time (RT)-qPCR and membrane potential was examined by flow cytometry.

RESULTS: We observed that EGCG inhibited in a dose-dependent manner both the planktonic growth and the biofilm formation of S. mutans. Significant reduction of S. mutans biofilm formation, DNA content, and EPS production was observed at 2.2-4.4 mg/ml EGCG. EGCG reduced the expression of gtfB, gtfC and ftf genes involved in EPS production, and the nox and sodA genes involved in the protection against oxidative stress. Moreover, EGCG caused an immediate change in membrane potential.

CONCLUSIONS: EGCG, a natural polyphenol, has a significant inhibitory effect on S. mutans dental biofilm formation and EPS production, and thus might be a potential drug in preventing dental caries.},
}

@article {pmid34525167,
year = {2021},
author = {Rivas, DP and Hedgecock, ND and Stebe, KJ and Leheny, RL},
title = {Dynamic and mechanical evolution of an oil-water interface during bacterial biofilm formation.},
journal = {Soft matter},
volume = {17},
number = {35},
pages = {8195-8210},
doi = {10.1039/d1sm00795e},
pmid = {34525167},
issn = {1744-6848},
mesh = {Bacteria ; *Biofilms ; Pseudomonas aeruginosa ; Rheology ; *Water ; },
abstract = {We present an experimental study combining particle tracking, active microrheology, and differential dynamic microscopy (DDM) to investigate the dynamics and rheology of an oil-water interface during biofilm formation by the bacteria Pseudomonas Aeruginosa PA14. The interface transitions from an active fluid dominated by the swimming motion of adsorbed bacteria at early age to an active viscoelastic system at late ages when the biofilm is established. The microrheology measurements using microscale magnetic rods indicate that the biofilm behaves as a viscoelastic solid at late age. The bacteria motility at the interface during the biofilm formation, which is characterized in the DDM measurements, evolves from diffusive motion at early age to constrained, quasi-localized motion at later age. Similarly, the mobility of passively moving colloidal spheres at the interface decreases significantly with increasing interface age and shows a dependence on sphere size after biofilm formation that is orders-of-magnitude larger than that expected in a homogeneous system in equilibrium. We attribute this anomalous size dependence to either length-scale-dependent rheology of the biofilm or widely differing effects of the bacteria activity on the motion of spheres of different sizes.},
}

Bacteria
*Biofilms
Pseudomonas aeruginosa
Rheology
*Water

@article {pmid34524972,
year = {2021},
author = {Gajdács, M and Kárpáti, K and Nagy, ÁL and Gugolya, M and Stájer, A and Burián, K},
title = {Association between biofilm-production and antibiotic resistance in Escherichia coli isolates: A laboratory-based case study and a literature review.},
journal = {Acta microbiologica et immunologica Hungarica},
volume = {},
number = {},
pages = {},
doi = {10.1556/030.2021.01487},
pmid = {34524972},
issn = {1588-2640},
abstract = {Bacteria can enhance their survival by attaching to inanimate surfaces or tissues, and presenting as multicellular communities encased in a protective extracellular matrix called biofilm. There has been pronounced interest in assessing the relationship between the antibiotic resistant phenotype and biofilm-production in clinically-relevant pathogens. The aim of the present paper was to provide additional experimental results on the topic, testing the biofilm-forming capacity of Escherichia coli isolates using in vitro methods in the context of their antibiotic resistance in the form of a laboratory case study, in addition to provide a comprehensive review of the subject. In our case study, a total of two hundred and fifty (n = 250) E. coli isolates, originating from either clean-catch urine samples (n = 125) or invasive samples (n = 125) were included. The colony morphology of isolates were recorded after 24h, while antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method. Biofilm-formation of the isolates was assessed with the crystal violet tube-adherence method. Altogether 57 isolates (22.8%) isolates were multidrug resistant (MDR), 89 isolates (35.6%) produced large colonies (>3 mm), mucoid variant colonies were produced in 131 cases (52.4%), and 108 (43.2%) were positive for biofilm formation. Biofilm-producers were less common among isolates resistant to third-generation cephalosporins and trimethoprim-sulfamethoxazole (P = 0.043 and P = 0.023, respectively). Biofilms facilitate a protective growth strategy in bacteria, ensuring safety against environmental stressors, components of the immune system and noxious chemical agents. Being an integral part of bacterial physiology, biofilm-formation is interdependent with the expression of other virulence factors (especially adhesins) and quorum sensing signal molecules. More research is required to allow for the full understanding of the interplay between the MDR phenotype and biofilm-production, which will facilitate the development of novel therapeutic strategies.},
}

@article {pmid34524634,
year = {2021},
author = {Liu, Z and Zhao, Z and Zeng, K and Xia, Y and Xu, W and Wang, R and Guo, J and Xie, H},
title = {Functional Immobilization of a Biofilm-Releasing Glycoside Hydrolase Dispersin B on Magnetic Nanoparticles.},
journal = {Applied biochemistry and biotechnology},
volume = {},
number = {},
pages = {},
pmid = {34524634},
issn = {1559-0291},
support = {31771032//National Natural Science Foundation of China/ ; 51911530153//National Natural Science Foundation of China/ ; },
abstract = {Dispersin B (DspB) is a member of glycoside hydrolase family 20 (GH20) and catalyzes degradation of biofilms forming by pathogenic bacteria such as Staphylococcus aureus. Magnetoreceptor (MagR) is a magnetic protein that can be used as a fusion partner for functionally immobilizing proteins on magnetic surfaces. In the present study, a recombinant protein DspB-MagR was constructed by fusing MagR to the C-terminus of DspB and expressed in Escherichia coli. Magnetic immobilization of purified DspB-MagR on magnetic core-shell structured Fe3O4@SiO2 nanoparticles was achieved and characterized by means of various techniques including SDS-PAGE, Fourier transform infrared spectroscopy, thermogravimetric analysis, zeta potential measurement, and scanning electron microscopy. It was evaluated the influence of temperature, pH, and storage time on the performance of immobilized DspB-MagR on Fe3O4@SiO2 nanoparticles. Removal of biofilms forming by Staphylococcus aureus and other medical sourced bacterial species was achieved by using Fe3O4@SiO2 nanoparticles loading with DspB-MagR. This work promoted potential applications of DspB and similar enzymes for medical purposes.},
}

@article {pmid34523997,
year = {2021},
author = {Staats, A and Burback, PW and Eltobgy, M and Parker, DM and Amer, AO and Wozniak, DJ and Wang, SH and Stevenson, KB and Urish, KL and Stoodley, P},
title = {Synovial Fluid-Induced Aggregation Occurs across Staphylococcus aureus Clinical Isolates and is Mechanistically Independent of Attached Biofilm Formation.},
journal = {Microbiology spectrum},
volume = {},
number = {},
pages = {e0026721},
doi = {10.1128/Spectrum.00267-21},
pmid = {34523997},
issn = {2165-0497},
support = {R01 GM124436/GM/NIGMS NIH HHS/United States ; },
abstract = {Rapid synovial fluid-induced aggregation of Staphylococcus aureus is currently being investigated as an important factor in the establishment of periprosthetic joint infections (PJIs). Pathogenic advantages of aggregate formation have been well documented in vitro, including recalcitrance to antibiotics and protection from host immune defenses. The objective of the present work was to determine the strain dependency of synovial fluid-induced aggregation by measuring the degree of aggregation of 21 clinical S. aureus isolates cultured from either PJI or bloodstream infections using imaging and flow cytometry. Furthermore, by measuring attached bacterial biomass using a conventional crystal violet assay, we assessed whether there is a correlation between the aggregative phenotype and surface-associated biofilm formation. While all of the isolates were stimulated to aggregate upon exposure to bovine synovial fluid (BSF) and human serum (HS), the extent of aggregation was highly variable between individual strains. Interestingly, the PJI isolates aggregated significantly more upon BSF exposure than those isolated from bloodstream infections. While we were able to stimulate biofilm formation with all of the isolates in growth medium, supplementation with either synovial fluid or human serum inhibited bacterial surface attachment over a 24 h incubation. Surprisingly, there was no correlation between the degree of synovial fluid-induced aggregation and quantity of surface-associated biofilm as measured by a conventional biofilm assay without host fluid supplementation. Taken together, our findings suggest that synovial fluid-induced aggregation appears to be widespread among S. aureus strains and mechanistically independent of biofilm formation. IMPORTANCE Bacterial infections of hip and knee implants are rare but devastating complications of orthopedic surgery. Despite a widespread appreciation of the considerable financial, physical, and emotional burden associated with the development of a prosthetic joint infection, the establishment of bacteria in the synovial joint remains poorly understood. It has been shown that immediately upon exposure to synovial fluid, the viscous fluid in the joint, Staphylococcus aureus rapidly forms aggregates which are resistant to antibiotics and host immune cell clearance. The bacterial virulence associated with aggregate formation is likely a step in the establishment of prosthetic joint infection, and as such, it has the potential to be a potent target of prevention. We hope that this work contributes to the future development of therapeutics targeting synovial fluid-induced aggregation to better prevent and treat these infections.},
}

@article {pmid34523579,
year = {2021},
author = {Fung, AHY and Rao, S and Ngan, WY and Sekoai, PT and Touyon, L and Ho, TM and Wong, KP and Habimana, O},
title = {Exploring the optimization of aerobic food waste digestion efficiency through the engineering of functional biofilm Bio-carriers.},
journal = {Bioresource technology},
volume = {341},
number = {},
pages = {125869},
doi = {10.1016/j.biortech.2021.125869},
pmid = {34523579},
issn = {1873-2976},
abstract = {The possibility of breaking down cellulose-rich food waste through biofilm engineering was investigated. Six previously isolated strains from naturally degrading fruits and vegetables, screened for biofilm-forming ability and cellulolytic activity, were selected to enrich a biocarrier seeding microbial consortium. The food waste model used in this study was cabbage which was aerobically digested under repeated water rinsing and regular effluent drainage. The engineered biocarrier biofilm's functionality was evaluated by tracing microbial succession following metagenomic sequencing, quantitative PCR, scanning electron microscopy, and cellulolytic activity before and after the digestion processes. The engineered microbial consortium demonstrated superior biofilm-forming ability on biocarriers than the original microbial consortium and generally displayed a higher cellulolytic activity. The presented study provides one of the few studies of food waste aerobic digestion using engineered biofilms. Insights presented in this study could help further optimize aerobic food waste digestion.},
}

@article {pmid34523551,
year = {2021},
author = {Song, Z and Su, X and Li, P and Sun, F and Dong, W and Zhao, Z and Wen, Z and Liao, R},
title = {Facial fabricated biocompatible homogeneous biocarriers involving biochar to enhance denitrification performance in an anoxic moving bed biofilm reactor.},
journal = {Bioresource technology},
volume = {341},
number = {},
pages = {125866},
doi = {10.1016/j.biortech.2021.125866},
pmid = {34523551},
issn = {1873-2976},
abstract = {Biochar prepared from pineapple peel was facially combined with polyurethane sponges for the first time to form homogeneous biocompatible biocarriers, which can enhance denitrification performance in an anoxic MBBR. The experiments showed that a higher NO3--N removal efficiency (96.24 ± 1.3%) and kinetic constant (0.26 h-1) were obtained in the MBBR employing these new biocarriers (B-MBBR), compared with a control MBBR with polyurethane sponges (C-MBBR). The attached and suspended biomass of the B-MBBR was increased by 47% and 26%, respectively. Biochar significantly enhanced the abundance of functional bacteria in terms of promoting biofilm (i.e., Leptonema), denitrifying bacteria (i.e., Thauera, Enterobacter and Pseudomonas) and electroactive bacteria (i.e., Geobacter) in the B-MBBR. Meanwhile, based on the content of coenzyme I (NADH) and denitrifying enzymes, biochar would also enhance electron transport activity for denitrification. Consequently, these facial prepared biocarriers are effective to enhance denitrification performance in MBBR with application significance.},
}

@article {pmid34523549,
year = {2021},
author = {Huang, T and Zhao, J and Hu, B and Zhao, J and Yuan, C},
title = {Effective restoration of partial nitritation and anammox biofilm process by short-term hydroxylamine dosing: Mechanism and microbial interaction.},
journal = {Bioresource technology},
volume = {341},
number = {},
pages = {125910},
doi = {10.1016/j.biortech.2021.125910},
pmid = {34523549},
issn = {1873-2976},
abstract = {The one-stage partial nitritation and anammox (PN-A) process frequently experiences deterioration from ammonium accumulation and nitrate build-up. In this study, hydroxylamine was dosed to restore the process from deterioration in a continuously aerated PN-A sequencing biofilm batch reactor, and the impact of hydroxylamine on the metabolism of PN-A process was studied. PN-A process was totally restored in 5 days via 10 mg N·L-1 hydroxylamine dosing, reducing nitrate-produced/ammonium-removed ratio from 28.5% to less than 11.0%. hydroxylamine dosing promoted biological production of nitric oxide and nitrous oxide and reduced the production of nitrate in the PN-A process. This study advanced the understanding of the metabolism versatility of hydroxylamine and nitric oxide as well as their function in interaction between aerobic ammonium oxidation bacteria and anaerobic ammonium oxidation bacteria, and proposed the potential application of hydroxylamine dosing in ammonium-contained wastewater treatment.},
}

@article {pmid34522977,
year = {2021},
author = {Sun, X and Xiang, J},
title = {Mechanism Underlying the Role of LuxR Family Transcriptional Regulator abaR in Biofilm Formation by Acinetobacter baumannii.},
journal = {Current microbiology},
volume = {},
number = {},
pages = {},
pmid = {34522977},
issn = {1432-0991},
support = {16ZR1420800//Natural Science Foundation of Shanghai Municipal Science and Technology Commission/ ; },
abstract = {Our study attempted to explore the mechanism underlying the role of LuxR family transcriptional regulator abaR in biofilm formation by Acinetobacter baumannii. The abaR gene was knocked out in ATCC 17978 strain using homologous recombination method. The growth curve and biofilm formation were measured in the wild type and abaR gene knockdown strains. Transcriptome sequencing was performed in the wild type and abaR gene knockdown strains following 8 h of culture. The growth curve in the abaR gene knockdown strain was similar to that of the wild-type strain. Biofilm formation significantly declined in the abaR gene knockdown strain at 8 and 48 h after culture. A total of 137 differentially expressed genes (DEGs) were obtained including 20 downregulated DEGs and 117 upregulated DEGs. Genes with differential expression were closely related to viral procapsid maturation (GO:0046797), acetoin catabolism (GO:0045150), carbon metabolism (ko01200), and the glycolysis/gluconeogenesis (ko00010)-related pathways. The results of the eight verified expression DEGs were consistent with the results predicted by bioinformatics. AbaR gene knockdown significantly affected biofilm formation by A. baumannii ATCC 17978 strain. The glycolysis/gluconeogenesis pathways were significantly dysregulated and induced by abaR gene knockdown in A. baumannii.},
}

@article {pmid34522106,
year = {2021},
author = {Yang, F and Liu, C and Ji, J and Cao, W and Ding, B and Xu, X},
title = {Molecular Characteristics, Antimicrobial Resistance, and Biofilm Formation of Pseudomonas aeruginosa Isolated from Patients with Aural Infections in Shanghai, China.},
journal = {Infection and drug resistance},
volume = {14},
number = {},
pages = {3637-3645},
pmid = {34522106},
issn = {1178-6973},
abstract = {Purpose: To investigate molecular characteristics, antimicrobial resistance, and biofilm formation ability of Pseudomonas aeruginosa strains isolated from patients with aural infections.

Methods: Isolates (n = 199) were collected from ear discharges of patients with aural infections from January 2019 to December 2020. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute guidelines. All isolates were subjected to multilocus sequence typing (MLST) with amplification and sequencing of seven housekeeping genes. Biofilm formation and eradication were quantitatively assessed in microtiter plates. Genes associated with biofilm formation and the quinolone-resistance-determining region (QRDR) of genes gyrA and parC were investigated using polymerase chain reaction amplification and sequencing.

Results: Of the 199 P. aeruginosa strains isolated, 109 (54.77%) were from females and 90 (45.23%) were from males. The isolates exhibited very low rates of resistance to most antibiotics tested, including piperacillin (1.51%), ceftazidime (0.50%), and imipenem (3.52%); however, the quinolones ciprofloxacin (80.40%) and levofloxacin (82.91%) were notable exceptions. The QRDR sequence results of the quinolone-resistant P. aeruginosa isolates showed Thr83Ile (n = 155) was the most common amino acid mutation in gyrA (n = 165), while Ser87Leu (n = 157) was widely detected in parC (n = 165). MLST analysis identified 34 sequence types (STs) with most isolates belonging to ST316 (73.87%). Almost all of the P. aeruginosa isolates (96.98%) produced biofilms and biofilm-forming genes algD (98.49%), pslD (96.98%), and pelF (96.48%) were highly prevalent.

Conclusion: The P. aeruginosa strains isolated from aural discharges in this study exhibited very low rates of resistance to most antibiotics tested, except for the resistance rates to quinolones, which were relatively high. The isolates also exhibited a strong biofilm formation ability and low susceptibility to eradication, indicating that more effective drugs and treatment methods are needed to combat these infections.},
}

@article {pmid34521475,
year = {2021},
author = {Du, C and Huo, X and Gu, H and Wu, D and Hu, Y},
title = {Acid resistance system CadBA is implicated in acid tolerance and biofilm formation and is identified as a new virulence factor of Edwardsiella tarda.},
journal = {Veterinary research},
volume = {52},
number = {1},
pages = {117},
pmid = {34521475},
issn = {1297-9716},
support = {2019CXTD413//Natural Science Foundation of Hunan Province/ ; },
abstract = {Edwardsiella tarda is a facultative intracellular pathogen in humans and animals. The Gram-negative bacterium is widely considered a potentially important bacterial pathogen. Adaptation to acid stress is important for the transmission of intestinal microbes, so the acid-resistance (AR) system is essential. However, the AR systems of E. tarda are totally unknown. In this study, a lysine-dependent acid resistance (LDAR) system in E. tarda, CadBA, was characterized and identified. CadB is a membrane protein and shares high homology with the lysine/cadaverine antiporter. CadA contains a PLP-binding core domain and a pyridoxal phosphate-binding motif. It shares high homology with lysine decarboxylase. cadB and cadA are co-transcribed under one operon. To study the function of the cadBA operon, isogenic cadA, cadB and cadBA deletion mutant strains TX01ΔcadA, TX01ΔcadB and TX01ΔcadBA were constructed. When cultured under normal conditions, the wild type strain and three mutants exhibited the same growth performance. However, when cultured under acid conditions, the growth of three mutants, especially TX01ΔcadA, were obviously retarded, compared to the wild strain TX01, which indicates the important involvement of the cadBA operon in acid resistance. The deletion of cadB or cadA, especially cadBA, significantly attenuated bacterial activity of lysine decarboxylase, suggesting the vital participation of cadBA operon in lysine metabolism, which is closely related to acid resistance. The mutations of cadBA operon enhanced bacterial biofilm formation, especially under acid conditions. The deletions of the cadBA operon reduced bacterial adhesion and invasion to Hela cells. Consistently, the deficiency of cadBA operon abated bacterial survival and replication in macrophages, and decreased bacterial dissemination in fish tissues. Our results also show that the expression of cadBA operon and regulator cadC were up-regulated upon acid stress, and CadC rigorously regulated the expression of cadBA operon, especially under acid conditions. These findings demonstrate that the AR CadBA system was a requisite for the resistance of E. tarda against acid stress, and played a critical role in bacterial infection of host cells and in host tissues. This is the first study about the acid resistance system of E. tarda and provides new insights into the acid-resistance mechanism and pathogenesis of E. tarda.},
}

@article {pmid34519903,
year = {2021},
author = {Kijkla, P and Wang, D and Mohamed, ME and Saleh, MA and Kumseranee, S and Punpruk, S and Gu, T},
title = {Efficacy of glutaraldehyde enhancement by D-limonene in the mitigation of biocorrosion of carbon steel by an oilfield biofilm consortium.},
journal = {World journal of microbiology & biotechnology},
volume = {37},
number = {10},
pages = {174},
pmid = {34519903},
issn = {1573-0972},
abstract = {Microbiologically influenced corrosion (MIC) is one of the major corrosion threats in the oil and gas industry. It is caused by environmental biofilms. Glutaraldehyde is a popular green biocide for mitigating biofilms and MIC. This work investigated the efficacy of glutaraldehyde enhancement by food-grade green chemical D-limonene in the biofilm prevention and MIC mitigation using a mixed-culture oilfield biofilm consortium. After 7 days of incubation at 37 °C in enriched artificial seawater in 125 mL anaerobic vials, the 100 ppm (w/w) glutaraldehyde + 200 ppm D-limonene combination treatment reduced the sessile cell counts on C1018 carbon steel coupons by 2.1-log, 1.7-log, and 2.3-log for sulfate reducing bacteria, acid producing bacteria, and general heterotrophic bacteria, respectively in comparison with the untreated control. The treatment achieved 68% weight loss reduction and 78% pit depth reduction. The 100 ppm glutaraldehyde + 200 ppm D-limonene combination treatment was found more effective in biofilm prevention and MIC mitigation than glutaraldehyde and D-limonene used individually. Electrochemical tests corroborated weight loss and pit depth data trends.},
}

@article {pmid34519743,
year = {2021},
author = {Fan, Q and Wang, C and Guo, R and Jiang, X and Li, W and Chen, X and Li, K and Hong, W},
title = {Step-by-step dual stimuli-responsive nanoparticles for efficient bacterial biofilm eradication.},
journal = {Biomaterials science},
volume = {},
number = {},
pages = {},
doi = {10.1039/d1bm01038g},
pmid = {34519743},
issn = {2047-4849},
abstract = {Biofilm-related bacterial infections are extremely resistant to antibiotics, mainly due to the impermeability of the intensive matrices, which allow the bacteria to survive antibiotic treatment. Herein, step-by-step dual stimuli-responsive azithromycin-loaded nanoparticles (CM/AZM@Tyr) was constructed for efficient biofilm eradication. CM/AZM@Tyr was prepared by the self-assembly of poly(ε-caprolactone)-polyethylene glycol-polyethylenimine (PCL-PEG-PEI) into cationic micelles and simultaneously encapsulated AZM into the hydrophobic core, which is further bound with cis-aconityl-D-tyrosine (CA-Tyr) through electrostatic interaction. Upon initial penetration, CM/AZM@Tyr could show step-by-step dual-response to the microenvironment of biofilms. Firstly, the CA-Tyr shell rapidly responded to the acidic microenvironment and released D-Tyr to disassemble the biofilm mass. Then, the exposed cationic CM/AZM micelles could bind firmly to the negatively-charged bacteria cell membrane. With the enzymolysis of the PCL core, the rapidly releasing AZM could kill the bacteria over the depth of biofilms. Massive accumulation was observed in the infected lungs of biofilms-associated lung infection mice after the i.v. injection of CM/Cy5.5@Tyr under the 3D mode of the in vivo Imaging System. Reduced bacterial burden and alleviated fibrosis in the infected lungs were also obtained after treatment with CM/AZM@Tyr mainly due to its intensive penetration in the biofilm and the orderly release of the biofilm dispersant and antimicrobial agents. In summary, this research developed an effective strategy for the treatment of blood-accessible biofilm-induced infections.},
}

@article {pmid34519474,
year = {2021},
author = {Zou, Y and Lu, K and Lin, Y and Wu, Y and Wang, Y and Li, L and Huang, C and Zhang, Y and Brash, JL and Chen, H and Yu, Q},
title = {Dual-Functional Surfaces Based on an Antifouling Polymer and a Natural Antibiofilm Molecule: Prevention of Biofilm Formation without Using Biocides.},
journal = {ACS applied materials & interfaces},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsami.1c10747},
pmid = {34519474},
issn = {1944-8252},
abstract = {Pathogenic biofilms formed on the surfaces of implantable medical devices and materials pose an urgent global healthcare problem. Although conventional antibacterial surfaces based on bacteria-repelling or bacteria-killing strategies can delay biofilm formation to some extent, they usually fail in long-term applications, and it remains challenging to eradicate recalcitrant biofilms once they are established and mature. From the viewpoint of microbiology, a promising strategy may be to target the middle stage of biofilm formation including the main biological processes involved in biofilm development. In this work, a dual-functional antibiofilm surface is developed based on copolymer brushes of 2-hydroxyethyl methacrylate (HEMA) and 3-(acrylamido)phenylboronic acid (APBA), with quercetin (Qe, a natural antibiofilm molecule) incorporated via acid-responsive boronate ester bonds. Due to the antifouling properties of the hydrophilic poly(HEMA) component, the resulting surface is able to suppress bacterial adhesion and aggregation in the early stages of contact. A few bacteria are eventually able to break through the protection of the anti-adhesion layer leading to bacterial colonization. In response to the resulting decrease in the pH of the microenvironment, the surface could then release Qe to interfere with the microbiological processes related to biofilm formation. Compared to bactericidal and anti-adhesive surfaces, this dual-functional surface showed significantly improved antibiofilm performance to prevent biofilm formation involving both Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus for up to 3 days. In addition, both the copolymer and Qe are negligibly cytotoxic, thereby avoiding possible harmful effects on adjacent normal cells and the risk of bacterial resistance. This dual-functional design approach addresses the different stages of biofilm formation, and (in accordance with the growth process of the biofilm) allows sequential activation of the functions without compromising the viability of adjacent normal cells. A simple and reliable solution may thus be provided to the problems associated with biofilms on surfaces in various biomedical applications.},
}

@article {pmid34517785,
year = {2021},
author = {Fu, R and Li, Z and Zhou, R and Li, C and Shao, S and Li, J},
title = {The mechanism of intestinal flora dysregulation mediated by intestinal bacterial biofilm to induce constipation.},
journal = {Bioengineered},
volume = {12},
number = {1},
pages = {6484-6498},
doi = {10.1080/21655979.2021.1973356},
pmid = {34517785},
issn = {2165-5987},
abstract = {To explore mechanism of intestinal flora dysregulation promoting constipation, 60 specific pathogen-free (SPF) mice were used as research objects and were treated with constipation population fecal fluid gavage and distilled water gavage. Then, relationship between intestinal dysregulation and constipation in mice with biofilm-mediated intestinal flora was investigated in vitro. The results showed that recombinant serotonin transporter (SERT) messenger ribonucleic acid (mRNA) level of the constipation population fecal fluid gavage group and the relative expression level of SERT mRNA were 1.61 ± 0.08 and 1.49 ± 0.06, which were higher markedly than those of distilled water group (P < 0.05). The level of 5-hydroxytryptamine (5-HT) in colonic tissue of the constipation population fecal fluid gavage group was 145.36 ± 14.12 ng/mL, and the expression level of 5-HT on the surface of epithelial cells of biofilm-positive colonic tissue was 20.11 ± 2.03, which were significantly lower than those of the distilled water group, with statistical significance (P < 0.05). Besides, the microbial sequencing of fecal flora indicated that The Akk and bacteroidetes ofconstipation population fecal fluid gavage group were higher hugely than those of distilled water group (P < 0.05).In conclusion, after the occurrence of constipation, the diversity of intestinal microflora decreased, and the probiotics reduced. Iintestinal microflora dysregulation would lead to increase of SERT expression level in defecation function and intestinal motility in mice, and the decrease of 5-HT, thereby changing the intestinal movement resulting in mucosal protective barrier damage,thereby causing changes in intestinal movement and the destruction of the intestinal mucosal protective barrier, which eventually resulted in constipation. The occurrence of constipation could be improved by regulating balance of intestinal flora, increasing the diversity of flora, and reducing the genus of opportunistic pathogens.},
}

@article {pmid34517574,
year = {2021},
author = {T, AV and Paramanantham, P and Sb, SL and Sharan, A and Alsaedi, MH and Dawoud, TMS and Syed, A and Siddhardha, B},
title = {Corrigendum to "Antimicrobial photodynamic activity of rose bengal conjugated multi walled carbon nanotubes against planktonic cells and biofilm of Escherichia coli" [Photodiagn. Photodyn. Ther. 24 (2021) 300-310].},
journal = {Photodiagnosis and photodynamic therapy},
volume = {35},
number = {},
pages = {102363},
doi = {10.1016/j.pdpdt.2021.102363},
pmid = {34517574},
issn = {1873-1597},
}

@article {pmid34517572,
year = {2021},
author = {Gao, Y and Mai, B and Wang, A and Li, M and Wang, X and Zhang, K and Liu, Q and Wei, S and Wang, P},
title = {Corrigendum to "Antimicrobial properties of a new type of photosensitizer derived from phthalocyanine against planktonic and biofilm forms of Staphylococcus aureus" [Photodiagn. Photodyn. Ther. 21 (2018) 316-326].},
journal = {Photodiagnosis and photodynamic therapy},
volume = {35},
number = {},
pages = {102337},
doi = {10.1016/j.pdpdt.2021.102337},
pmid = {34517572},
issn = {1873-1597},
}

@article {pmid34516283,
year = {2021},
author = {Kassety, S and Katharios-Lanwermeyer, S and O'Toole, GA and Nadell, CD},
title = {Differential surface competition and biofilm invasion strategies of Pseudomonas aeruginosa PA14 and PAO1.},
journal = {Journal of bacteriology},
volume = {},
number = {},
pages = {JB0026521},
doi = {10.1128/JB.00265-21},
pmid = {34516283},
issn = {1098-5530},
abstract = {Pseudomonas aeruginosa strains PA14 and PAO1 are among the two best characterized model organisms used to study the mechanisms of biofilm formation, while also representing two distinct lineages of P. aeruginosa. Previous work has shown that PA14 and PAO1 use different strategies for surface colonization; they also have different extracellular matrix composition and different propensities to disperse from biofilms back into the planktonic phase surrounding them. We expand on this work here by exploring the consequences of these different biofilm production strategies during direct competition. Using differentially labeled strains and microfluidic culture methods, we show that PAO1 can outcompete PA14 in direct competition during early colonization and subsequent biofilm growth, that they can do so in constant and perturbed environments, and that this advantage is specific to biofilm growth and requires production of the Psl polysaccharide. In contrast, the P. aeruginosa PA14 is better able to invade pre-formed biofilms and is more inclined to remain surface-associated under starvation conditions. These data together suggest that while P. aeruginosa PAO1 and PA14 are both able to effectively colonize surfaces, they do so in different ways that are advantageous under different environmental settings. Importance Recent studies indicate that P. aeruginosa PAO1 and PA14 use distinct strategies to initiate biofilm formation. We investigated whether their respective colonization and matrix secretion strategies impact their ability to compete under different biofilm-forming regimes. Our work shows that these different strategies do indeed impact how these strains fair in direct competition: PAO1 dominates during colonization of a naïve surface, while PA14 is more effective in colonizing a pre-formed biofilm. These data suggest that even for very similar microbes there can be distinct strategies to successfully colonize and persist on surfaces during the biofilm life cycle.},
}

@article {pmid34516241,
year = {2021},
author = {Singulani, JL and Oliveira, LT and Ramos, MD and Fregonezi, NF and Gomes, PC and Galeane, MC and Palma, MS and Fusco Almeida, AM and Mendes Giannini, MJS},
title = {The antimicrobial peptide MK58911-NH2 acts on planktonic, biofilm and intramacrophage cells of Cryptococcus neoformans.},
journal = {Antimicrobial agents and chemotherapy},
volume = {},
number = {},
pages = {AAC0090421},
doi = {10.1128/AAC.00904-21},
pmid = {34516241},
issn = {1098-6596},
abstract = {Cryptococcosis is associated with high rates of morbidity and mortality, especially in AIDS patients. Its treatment is carried out by combining amphotericin B and azoles or flucytosine, which cause unavoidable toxicity issues to the host. Thus, the urgency in obtaining new antifungals drives the search for antimicrobial peptides (AMPs). This study aimed to extend the understanding of the mechanism of action of an AMP analog from wasps peptide toxins, MK58911-NH2, on Cryptococcus neoformans. It was also evaluated if MK58911-NH2 can act on cryptococcal cells in macrophages, biofilms, and an immersion zebrafish model of infection. Finally, we investigated the structure-antifungal action and the toxicity relation of MK58911-NH2 fragments and a derivative of this peptide (MH58911-NH2). The results demonstrated that MK58911-NH2 did not alter the fluorescence intensity of cell wall - binding dye calcofluor or capsule- binding dye 18b7 antibody-FITC of C. neoformans, but rather reduced the number and size of fungal cells. This activity reduced the fungal burden of C. neoformans both in macrophages and in zebrafish embryos as well as within biofilms. Three fragments of the MK58911-NH2 peptide showed no activity against Cryptococcus or toxicity in lung cells. The derivative peptide MH58911-NH2, in which the lysine residues of MK58911-NH2 were replaced by histidine, reduced the activity against extracellular and intracellular C. neoformans. On the other hand, it was active against biofilm, and reducing toxicity. In summary, the results showed that peptide MK58911-NH2 could be a promising agent against cryptococcosis. The work also opens a perspective for the verification of the antifungal activity of other derivatives.},
}

@article {pmid34512560,
year = {2021},
author = {Thieme, L and Hartung, A and Tramm, K and Graf, J and Spott, R and Makarewicz, O and Pletz, MW},
title = {Adaptation of the Start-Growth-Time Method for High-Throughput Biofilm Quantification.},
journal = {Frontiers in microbiology},
volume = {12},
number = {},
pages = {631248},
pmid = {34512560},
issn = {1664-302X},
abstract = {Colony forming unit (CFU) determination by agar plating is still regarded as the gold standard for biofilm quantification despite being time- and resource-consuming. Here, we propose an adaption of the high-throughput Start-Growth-Time (SGT) method from planktonic to biofilm analysis, which indirectly quantifies CFU/mL numbers by evaluating regrowth curves of detached biofilms. For validation, the effect of dalbavancin, rifampicin and gentamicin against mature biofilms of Staphylococcus aureus and Enterococcus faecium was measured by accessing different features of the viability status of the cell, i.e., the cultivability (conventional agar plating), growth behavior (SGT) and metabolic activity (resazurin assay). SGT correlated well with the resazurin assay for all tested antibiotics, but only for gentamicin and rifampicin with conventional agar plating. Dalbavancin treatment-derived growth curves showed a compared to untreated controls significantly slower increase with reduced cell doubling times and reduced metabolic rate, but no change in CFU numbers was observed by conventional agar plating. Here, unspecific binding of dalbavancin to the biofilm interfered with the SGT methodology since the renewed release of dalbavancin during detachment of the biofilms led to an unintended antimicrobial effect. The application of the SGT method for anti-biofilm testing is therefore not suited for antibiotics which stick to the biofilm and/or to the bacterial cell wall. Importantly, the same applies for the well-established resazurin method for anti-biofilm testing. However, for antibiotics which do not bind to the biofilm as seen for gentamicin and rifampicin, the SGT method presents a much less labor-intensive method suited for high-throughput screening of anti-biofilm compounds.},
}

@article {pmid34511397,
year = {2021},
author = {Segundo Zaragoza, C and López Ortiz, I and Contreras Caro Del Castillo, DA and Domínguez Hernández, YM and Rodríguez García, JA},
title = {Characterization, enzymatic activity and biofilm formation of Candida species isolated from goat milk.},
journal = {Revista iberoamericana de micologia},
volume = {},
number = {},
pages = {},
doi = {10.1016/j.riam.2021.06.001},
pmid = {34511397},
issn = {2173-9188},
abstract = {BACKGROUND: Data regarding yeast microbiota in goat milk is scarce.

AIMS: To isolate and identify species of the genus Candida in milk samples from clinically healthy goats, and evaluate their enzymatic activity and biofilm formation.

METHODS: 1092 milk samples from clinically healthy goats were collected and processed. The yeast isolates were identified by phenotypic, methods and their enzymatic activity (phospholipase, hemolysin and protease) and biofilm formation evaluated.

RESULTS: We obtained 221 Candida isolates belonging to six species: Candida kefyr (35.7%), Candida guilliermondii (33%), Candida famata (23.5%), Candida glabrata (5.9%), Candida albicans (1.35%) and Candida parapsilosissensu lato (0.45%). Protease activity was detected in all Candida species while hemolysin activity was only present in C. kefyr, C. guilliermondii, C. famata and C. albicans. Only C. albicans showed phospholipase activity. With the exception of C. parapsilosis sensu lato, all Candida species formed biofilm, with 60.19% of the isolates being poor producers, 9.93% moderate producers, and 1.35% strong producers.

CONCLUSIONS: The milk of clinically healthy goats contains several species of the genus Candida that could play a role as opportunistic pathogens in mastitis.},
}

@article {pmid34511035,
year = {2021},
author = {Zhang, W and Li, H and Zhao, N and Luo, X and Liu, S and Bao, A and Chen, Y and Wang, H and Wang, J and Wang, J},
title = {Lactobacillus johnsonii BS15 combined with abdominal massage on intestinal permeability in rats with nonalcoholic fatty liver and cell biofilm repair.},
journal = {Bioengineered},
volume = {12},
number = {1},
pages = {6354-6363},
doi = {10.1080/21655979.2021.1954134},
pmid = {34511035},
issn = {2165-5987},
abstract = {This study aimed to analyze the effect of lactobacillus johnsonii BS15 (isolation of homemade yogurt from Ahu Hongyuan Grassland) combined with abdominal massage on intestinal permeability in rats with nonalcoholic fatty liver disease (NAFLD) and cell biofilm repair. Forty-five rats were divided randomly into five groups, four of which were fed with high-fat diet to establish NAFLD models. According to the treatment methods, they were grouped into group A (lactic acid bacteria feeding), group B (abdominal massage), group A + B (a combination of the two methods), model group (distilled water feeding), and normal group (distilled water feeding). Then, the pathological indexes of liver and intestinal permeability were observed. FITC-Dextran content of the model group elevated markedly compared with normal group (P < 0.01), indicating that the intestinal permeability of NAFLD rats fed with high-fat diet increased. The intestinal permeability of groups A, B, and A + B was lower sharply than that of model group (P < 0.01), and the effect of group A + B was the most obvious. HE staining of liver tissues showed that combined treatment could improve structural changes in liver cells caused by modeling and restore the normal structure of intestinal cells. Lactobacillus combined with abdominal massage was better than two treatments alone, further promoting the permeability of intestinal mucosa in NAFLD rats and repair biofilm of hepatocytes. The results initially verified the intervention effect of abdominal massage on intestinal mucosal permeability, and further revealed the mechanism of abdominal massage in treatment of NAFLD by improving intestinal mucosal barrier permeability.},
}

@article {pmid34510307,
year = {2021},
author = {Gomaa, OM and Abd El Kareem, H and Selim, N},
title = {Nitrate modulation of Bacillus sp. biofilm components: a proposed model for sustainable bioremediation.},
journal = {Biotechnology letters},
volume = {},
number = {},
pages = {},
pmid = {34510307},
issn = {1573-6776},
abstract = {The presence of different pollutants in wastewater hinder microbial growth, compromise enzymatic activity or compete for electrons required for bioremediation pathway. Therefore, there is a need to use a single microorganism that is capable of tolerating different toxic compounds and can perform simultaneous bioremediation. In the present study, nitrate reducing bacteria capable of decolorizing azo dye was identified as Bacillus subtillis sp. DN using protein profiling, morphological and biochemical tests X-ray diffraction pattern, Raman spectroscopy and cyclic voltammetry confirm that the bacterium under study possesses membrane-bound nitrate reductase and that is capable of direct electron transfer. The addition of nitrate concentrations (0-50 mM) resulted in increased biofilm formation with variable exopolysaccharides, protein, and eDNA. Fourier Transform Infrared spectrum revealed the presence of a biopolymer at high nitrate concentrations. Effective capacitance and conductivity of the cells grown in different nitrate concentrations suggest changes in the relative position of polar groups, their relative orientation and permeability of cell membrane as detected by dielectric spectroscopy. The increase in biofilm shifted the removal of the azo dye from biodegradation to bioadsorption. Our results indicate that nitrate modulates biofilm components. Bacillus sp. DN granular biofilm can be used for simultaneous nitrate and azo dye removal from wastewater.},
}

@article {pmid34509685,
year = {2021},
author = {Garcia, BA and Panariello, BHD and Freitas-Pontes, KM and Duarte, S},
title = {Candida biofilm matrix as a resistance mechanism against photodynamic therapy.},
journal = {Photodiagnosis and photodynamic therapy},
volume = {36},
number = {},
pages = {102525},
doi = {10.1016/j.pdpdt.2021.102525},
pmid = {34509685},
issn = {1873-1597},
abstract = {BACKGROUND: Antimicrobial photodynamic therapy (aPDT) efficiency on Candida albicans is recognized in free-floating cultures. Though, the lack of aPDT effectiveness against C. albicans organized in biofilms is still unclear. This study aimed to explore the role of the extracellular matrix (ECM) in the protection against aPDT in C. albicans biofilms.

METHODS: C. albicans SN 425 wild-type and two mutant strains CNJ 2302; Δ/Δefg1 and CJN 2330; Δ/Δtec1 (ECM deficient) were used. Biofilms were grown on 24-well plates and exposed twice-daily to aPDT with 44 μM toluidine blue-O (TBO) for 5 min followed by red light (635 nm) for 1 min (87.6 J/cm²) or 2 min (175.2 J/cm2). Application of just TBO, light, 0.12% chlorhexidine, and ultrapure water were used as controls. After 48 h, biofilms were assessed for dry-weight (DW), colony forming units (CFU), extracellular DNA (eDNA), soluble and insoluble protein (SP/IP), water-insoluble (alkali-soluble) polysaccharide (ASP), water-soluble polysaccharides (WSP), and confocal scanning laser microscopy.

RESULTS: The strains with ECM deficient were affected by aPDT. For the mutant strain Δ/Δefg1, aPDT significantly reduced CFU, ASP, DW, eDNA, WSP and IP when compared to NC (p<0.001) and for the Δ/Δtec1, aPDT significantly reduced CFU, eDNA, IP and SP. Whereas CFU, DW, ASP of the wild-type strain biofilms were not reduced (p>0.05).

CONCLUSIONS: C. albicans strains with reduced ECM compounds were more sensitive to aPDT suggesting that the ECM may have a significant protection role from aPDT in C. albicans biofilms.},
}

@article {pmid34507762,
year = {2021},
author = {Fernández-Gómez, P and Figueredo, A and López, M and González-Raurich, M and Prieto, M and Alvarez-Ordóñez, A},
title = {Heterogeneity in biofilm formation and identification of biomarkers of strong biofilm formation among field isolates of Pseudomonas spp.},
journal = {Food research international (Ottawa, Ont.)},
volume = {148},
number = {},
pages = {110618},
doi = {10.1016/j.foodres.2021.110618},
pmid = {34507762},
issn = {1873-7145},
abstract = {The biofilm formation ability of a collection of thirty-three Pseudomonas spp. isolates from food processing facilities was investigated in order to find biomarkers of strong biofilm production, a characteristic that can determine persistence in food processing environments. The strains were classified according to the colony pigmentation on solid media as green, brown or not pigmented. The biofilm production on stainless steel and polystyrene was assessed by spectrometric determination of the fixed crystal violet, and the biofilm formed on glass, through confocal laser scanning microscopy. Besides, pyoverdine production, catalase activity, RpoS status and cellular hydrophobicity were also monitored. A significantly higher biofilm production level on stainless steel and polystyrene was observed for green-pigmented strains as compared to brown or not pigmented strains. The influence of iron availability on biofilm formation on stainless steel was studied through the addition of the iron scavenger 2,2-bipyridine resulting in a decrease of 40 % in biofilm formation for the not pigmented strains. For most of the potential biomarkers studied (i.e., pyoverdine production, catalase activity, cellular hydrophobicity), the phenotypic heterogeneity observed among strains was mainly dependent on the Pseudomonas species and no strong associations with the biofilm formation capacity were detected. However, the green colony pigmentation on solid media showed good potential as a biomarker of strong biofilm formation on stainless steel and polystyrene both in P. aeruginosa and Pseudomonas spp.},
}

@article {pmid34507740,
year = {2021},
author = {Hossain, MI and Mizan, MFR and Roy, PK and Nahar, S and Toushik, SH and Ashrafudoulla, M and Jahid, IK and Lee, J and Ha, SD},
title = {Listeria monocytogenes biofilm inhibition on food contact surfaces by application of postbiotics from Lactobacillus curvatus B.67 and Lactobacillus plantarum M.2.},
journal = {Food research international (Ottawa, Ont.)},
volume = {148},
number = {},
pages = {110595},
doi = {10.1016/j.foodres.2021.110595},
pmid = {34507740},
issn = {1873-7145},
abstract = {Owing to their preservative and antimicrobial effects, postbiotics (metabolic byproducts of probiotics) are promising natural components for the food industry. Therefore, the present study aimed to investigate the efficacy of postbiotics collected from isolated Lactobacillus curvatus B.67 and Lactobacillus plantarum M.2 against Listeria monocytogenes pathogens in planktonic cells, motility, and biofilm states. The analysis of the metabolite composition of the postbiotics revealed various organic acids, along with a few well-known bacteriocin-encoding genes with potential antimicrobial effects. Postbiotics maintained their residual antimicrobial activity over the pH range 1-6 but lost all activity at neutral pH (pH 7). Full antimicrobial activity (100%) was observed during heat treatment, even under the autoclaving condition.Minimum inhibitory concentration (MICs) of L. curvatus B.67 and L. plantarum M.2 against L. monocytogenes were 80 and 70 mg/mL, respectively. However, four sub-MICs of the postbiotics (1/2, 1/4, 1/8, and 1/16 MIC) were tested for inhibition efficacy against L. monocytogenes during different experiment in this study. Swimming motility, biofilm formation, and expression levels of target genes related to biofilm formation, virulence, and quorum-sensing were significantly inhibited with increasing postbiotics concentration. Postbiotics from L. plantarum M.2 exhibited a higher inhibitory effect than the postbiotics from L. curvatus B.67. Nonetheless, both these postbiotics from Lactobacillus spp. could be used as effective bio-interventions for controlling L. monocytogenes biofilm in the food industry.},
}

@article {pmid34506798,
year = {2021},
author = {Oh, JH and Park, J and Park, Y},
title = {Anti-biofilm and anti-inflammatory effects of Lycosin-II isolated from spiders against multi-drug resistant bacteria.},
journal = {Biochimica et biophysica acta. Biomembranes},
volume = {},
number = {},
pages = {183769},
doi = {10.1016/j.bbamem.2021.183769},
pmid = {34506798},
issn = {1879-2642},
abstract = {Currently, multidrug-resistant bacteria are rapidly increasing worldwide because of the misuse or overuse of antibiotics. In particular, few options exist for treating infections caused by long-persisting oxacillin-resistant strains and recently proliferating carbapenem-resistant strains. Therefore, alternative treatments are urgently needed. The antimicrobial peptide (AMP) Lycosin-II is a peptide consisting of 21 amino acids isolated from the venom of the spider Lycosa singoriensis. Lycosin-II showed strong antibacterial activity and biofilm inhibition effects against gram-positive and gram-negative bacteria including oxacillin-resistant Staphylococcus aureus (S. aureus) and meropenem-resistant Pseudomonas aeruginosa (P. aeruginosa) isolated from patients. In addition, Lycosin-II was not cytotoxic against human foreskin fibroblast Hs27 or hemolytic against sheep red blood cells at the concentration of which exerted antibacterial activity. The mechanism of action of Lycosin-II involves binding to lipoteichoic acid and lipopolysaccharide of gram-positive and gram-negative bacterial membranes, respectively, to destroy the bacterial membrane. Moreover, Lycosin-II showed anti-inflammatory effects by inhibiting the expression of pro-inflammatory cytokines that are increased during bacterial infection in Hs27 cells. These results suggest that Lycosin-II can serve as a therapeutic agent against infections with multidrug-resistant strains.},
}

@article {pmid34505216,
year = {2021},
author = {Gross, M and Ashqar, F and Sionov, RV and Friedman, M and Eliashar, R and Zaks, B and Gati, I and Duanis-Assaf, D and Feldman, M and Steinberg, D},
title = {Sustained release varnish containing chlorhexidine for prevention of Streptococcus mutans biofilm formation on voice prosthesis surface: an in vitro study.},
journal = {International microbiology : the official journal of the Spanish Society for Microbiology},
volume = {},
number = {},
pages = {},
pmid = {34505216},
issn = {1618-1905},
support = {20180025//The Israeli Cancer Association/ ; },
abstract = {OBJECTIVES: In this study, we aimed to develop a novel, sustained release varnish (SRV) for voice prostheses (VP) releasing chlorhexidine (CHX), for the prevention of biofilm formation caused by the common oral bacteria Streptococcus mutans on VP surfaces.

METHODS: This study was performed in an in vitro model as a step towards future in vivo trials. VPs were coated with a SRV containing CHX (SRV-CHX) or SRV alone (placebo-SRV) that were daily exposed to S. mutans. The polymeric materials of SRV were composed of ethylcellulose and PEG-400. Biofilm formation was assessed by DNA quantification (qPCR), crystal violet staining, confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), and kinetics experiments.

RESULTS: The amount of DNA in the biofilms formed by S. mutans on VP surfaces coated once with SRV-CHX (1.024 ± 0.218 ng DNA/piece) was 58.5 ± 8.8% lower than that of placebo-SRV-coated VPs (2.465 ± 0.198 ng DNA/piece) after a 48-h exposure to S. mutans (p = 0.038). Reduced biofilm mass on SRV-CHX-coated VPs was visually confirmed by CLSM and SEM. CV staining of SRV-CHX single-coated VPs that have been exposed to S. mutans nine times showed a 98.1 ± 0.2% reduction in biofilm mass compared to placebo-SRV-coated VPs (p = 0.003). Kinetic experiments revealed that SRV-CHX triple-coated VPs could delay bacterial growth for 23 days.

CONCLUSIONS: Coating VPs with SRV-CHX has an inhibitory effect on biofilm formation and prevents bacterial growth in their vicinities. This study is a proof-of-principle that paves the way for developing new clinical means for reducing both VPs' bacterial biofilm formation and device failure.},
}

@article {pmid34504987,
year = {2021},
author = {Arias, SL and Brito, IL},
title = {Biophysical determinants of biofilm formation in the gut.},
journal = {Current opinion in biomedical engineering},
volume = {18},
number = {},
pages = {},
pmid = {34504987},
issn = {2468-4511},
support = {R33 CA235302/CA/NCI NIH HHS/United States ; },
abstract = {The gastrointestinal (GI) tract harbors the most complex microbial ecosystem in the human body. The mucosal layer that covers the GI tract serves as a polymer-based defensive barrier that prevents the microbiome from reaching the epithelium and disseminating inside the body. Colonization of the mucus may result in the formation of structured polymicrobial communities or biofilms, a hallmark in pathologies such as colorectal cancer, inflammatory bowel disease, and chronic gut wounds. However, the mechanisms by which multispecies biofilms establish on the gut mucosa is unknown. Whether mucus-associated biofilms exist as part of a healthy mucosal barrier is still debated. Here, we discuss the impact that diet and microbial-derived polymers has on mucus structure and microcolony formation and highlight relevant biophysical forces that further shape nascent biofilms.},
}

@article {pmid34504737,
year = {2021},
author = {Janssen, K and Low, SL and Wang, Y and Mu, QY and Bierbaum, G and Gee, CT},
title = {Elucidating biofilm diversity on water lily leaves through 16S rRNA amplicon analysis: Comparison of four DNA extraction kits.},
journal = {Applications in plant sciences},
volume = {9},
number = {8},
pages = {e11444},
pmid = {34504737},
issn = {2168-0450},
abstract = {Premise: Within a broader study on leaf fossilization in freshwater environments, a long-term study on the development and microbiome composition of biofilms on the foliage of aquatic plants has been initiated to understand how microbes and biofilms contribute to leaf decay and preservation. Here, water lily leaves are employed as a study model to investigate the relationship between bacterial microbiomes, biodegradation, and fossilization. We compare four DNA extraction kits to reduce biases in interpretation and to identify the most suitable kit for the extraction of DNA from bacteria associated with biofilms on decaying water lily leaves for 16S rRNA amplicon analysis.

Methods: We extracted surface-associated DNA from Nymphaea leaves in early stages of decay at two water depth levels using four commercially available kits to identify the most suitable protocol for bacterial extraction, applying a mock microbial community standard to enable a reliable comparison of the kits.

Results: Kit 4, the FastDNA Spin Kit for Soil, resulted in high DNA concentrations with better quality and yielded the most accurate depiction of the mock community. Comparison of the leaves at two water depths showed no significant differences in community composition.

Discussion: The success of Kit 4 may be attributed to its use of bead beating with a homogenizer, which was more efficient in the lysis of Gram-positive bacteria than the manual vortexing protocols used by the other kits. Our results show that microbial composition on leaves during early decay remains comparable and may change only in later stages of decomposition.},
}

@article {pmid34503352,
year = {2021},
author = {Sharma, A and Vashistt, J and Shrivastava, R},
title = {Response surface modeling integrated microtiter plate assay for Mycobacterium fortuitum biofilm quantification.},
journal = {Biofouling},
volume = {},
number = {},
pages = {1-14},
doi = {10.1080/08927014.2021.1974846},
pmid = {34503352},
issn = {1029-2454},
abstract = {In this study, the effects of agitation, temperature, and pH on biofilm formation by Mycobacterium fortuitum were studied and quantified through response surface modeling. The microtiter plate assay was optimized to achieve conditions favoring maximum mycobacterial biofilm quantification. Optical density (OD) measurement using a crystal violet assay was performed to estimate the amount of biofilm formed. Response surface methodology (RSM) results revealed an R2 value of 96.18%, exhibiting a maximum OD of 2.119 (λ570 nm) at a temperature of 37 °C and pH 7.0, under a static environment. The conditions were experimentally validated. Statistically significant results showed that the maximum biofilm was produced 96 h after mycobacterial inoculation. Thus, the results provide a basis for using RSM as an efficient optimization method for M. fortuitum biofilm assays. This approach can also be incorporated into strategies for screening anti-biofilm compounds, synthetic chemicals, drugs, or inhibitors against pathogenic mycobacteria.},
}

@article {pmid34502269,
year = {2021},
author = {Pusparajah, P and Letchumanan, V and Law, JW and Ab Mutalib, NS and Ong, YS and Goh, BH and Tan, LT and Lee, LH},
title = {Streptomyces sp.-A Treasure Trove of Weapons to Combat Methicillin-Resistant Staphylococcus aureus Biofilm Associated with Biomedical Devices.},
journal = {International journal of molecular sciences},
volume = {22},
number = {17},
pages = {},
pmid = {34502269},
issn = {1422-0067},
support = {FRGS/1/2019/SKK08/MUSM/02/7//Ministry of Education - Fundamental Research Grant Scheme/ ; ECR-000014//JCSMHS Early Career Researcher Grant 2021/ ; },
abstract = {Biofilms formed by methicillin-resistant S. aureus (MRSA) are among the most frequent causes of biomedical device-related infection, which are difficult to treat and are often persistent and recurrent. Thus, new and effective antibiofilm agents are urgently needed. In this article, we review the most relevant literature of the recent years reporting on promising anti-MRSA biofilm agents derived from the genus Streptomyces bacteria, and discuss the potential contribution of these newly reported antibiofilm compounds to the current strategies in preventing biofilm formation and eradicating pre-existing biofilms of the clinically important pathogen MRSA. Many efforts are evidenced to address biofilm-related infections, and some novel strategies have been developed and demonstrated encouraging results in preclinical studies. Nevertheless, more in vivo studies with appropriate biofilm models and well-designed multicenter clinical trials are needed to assess the prospects of these strategies.},
}

@article {pmid34499698,
year = {2021},
author = {Taggart, MG and Snelling, WJ and Naughton, PJ and La Ragione, RM and Dooley, JSG and Ternan, NG},
title = {Biofilm regulation in Clostridioides difficile: Novel systems linked to hypervirulence.},
journal = {PLoS pathogens},
volume = {17},
number = {9},
pages = {e1009817},
pmid = {34499698},
issn = {1553-7374},
abstract = {Clostridiodes difficile (C. difficile) was ranked an "urgent threat" by the Centers for Disease Control and Prevention (CDC) in 2019. C. difficile infection (CDI) is the most common healthcare-associated infection (HAI) in the United States of America as well as the leading cause of antibiotic-associated gastrointestinal disease. C. difficile is a gram-positive, rod-shaped, spore-forming, anaerobic bacterium that causes infection of the epithelial lining of the gut. CDI occurs most commonly after disruption of the human gut microflora following the prolonged use of broad-spectrum antibiotics. However, the recurrent nature of this disease has led to the hypothesis that biofilm formation may play a role in its pathogenesis. Biofilms are sessile communities of bacteria protected from extracellular stresses by a matrix of self-produced proteins, polysaccharides, and extracellular DNA. Biofilm regulation in C. difficile is still incompletely understood, and its role in disease recurrence has yet to be fully elucidated. However, many factors have been found to influence biofilm formation in C. difficile, including motility, adhesion, and hydrophobicity of the bacterial cells. Small changes in one of these systems can greatly influence biofilm formation. Therefore, the biofilm regulatory system would need to coordinate all these systems to create optimal biofilm-forming physiology under appropriate environmental conditions. The coordination of these systems is complex and multifactorial, and any analysis must take into consideration the influences of the stress response, quorum sensing (QS), and gene regulation by second messenger molecule cyclic diguanosine monophosphate (c-di-GMP). However, the differences in biofilm-forming ability between C. difficile strains such as 630 and the "hypervirulent" strain, R20291, make it difficult to assign a "one size fits all" mechanism to biofilm regulation in C. difficile. This review seeks to consolidate published data regarding the regulation of C. difficile biofilms in order to identify gaps in knowledge and propose directions for future study.},
}

@article {pmid34497894,
year = {2021},
author = {Fasiku, VO and Omolo, CA and Devnarain, N and Ibrahim, UH and Rambharose, S and Faya, M and Mocktar, C and Singh, SD and Govender, T},
title = {Chitosan-Based Hydrogel for the Dual Delivery of Antimicrobial Agents Against Bacterial Methicillin-Resistant Staphylococcus aureus Biofilm-Infected Wounds.},
journal = {ACS omega},
volume = {6},
number = {34},
pages = {21994-22010},
pmid = {34497894},
issn = {2470-1343},
abstract = {Chronic wound infections caused by antibiotic-resistant bacteria have become a global health concern. This is attributed to the biofilm-forming ability of bacteria on wound surfaces, thus enabling their persistent growth. In most cases, it leads to morbidity and in severe cases mortality. Current conventional approaches used in the treatment of biofilm wounds are proving to be ineffective due to limitations such as the inability to penetrate the biofilm matrix; hence, biofilm-related wounds remain a challenge. Therefore, there is a need for more efficient alternate therapeutic interventions. Hydrogen peroxide (HP) is a known antibacterial/antibiofilm agent; however, prolonged delivery has been challenging due to its short half-life. In this study, we developed a hydrogel for the codelivery of HP and antimicrobial peptides (Ps) against bacteria, biofilms, and wound infection associated with biofilms. The hydrogel was prepared via the Michael addition technique, and the physiochemical properties were characterized. The safety, in vitro, and in vivo antibacterial/antibiofilm activity of the hydrogel was also investigated. Results showed that the hydrogel is biosafe. A greater antibacterial effect was observed with HP-loaded hydrogels (CS-HP; hydrogel loaded with HP and CS-HP-P; hydrogel loaded with HP and peptide) when compared to HP as seen in an approximately twofold and threefold decrease in minimum inhibitory concentration values against methicillin-resistant Staphylococcus aureus (MRSA) bacteria, respectively. Similarly, both the HP-releasing hydrogels showed enhanced antibiofilm activity in the in vivo study in mice models as seen in greater wound closure and enhanced wound healing in histomorphological analysis. Interestingly, the results revealed a synergistic antibacterial/antibiofilm effect between HP and P in both in vitro and in vivo studies. The successfully prepared HP-releasing hydrogels showed the potential to combat bacterial biofilm-related infections and enhance wound healing in mice models. These results suggest that the HP-releasing hydrogels may be a superior platform for eliminating bacterial biofilms without using antibiotics in the treatment of chronic MRSA wound infections, thus improving the quality of human health.},
}

@article {pmid34496717,
year = {2021},
author = {Martins Antunes de Melo, WC and Celiešiūtė-Germanienė, R and Šimonis, P and Stirkė, A},
title = {Antimicrobial photodynamic therapy (aPDT) for biofilm treatments. Possible synergy between aPDT and pulsed electric fields.},
journal = {Virulence},
volume = {12},
number = {1},
pages = {2247-2272},
pmid = {34496717},
issn = {2150-5608},
abstract = {Currently, microbial biofilms have been the cause of a wide variety of infections in the human body, reaching 80% of all bacterial and fungal infections. The biofilms present specific properties that increase the resistance to antimicrobial treatments. Thus, the development of new approaches is urgent, and antimicrobial photodynamic therapy (aPDT) has been shown as a promising candidate. aPDT involves a synergic association of a photosensitizer (PS), molecular oxygen and visible light, producing highly reactive oxygen species (ROS) that cause the oxidation of several cellular components. This therapy attacks many components of the biofilm, including proteins, lipids, and nucleic acids present within the biofilm matrix; causing inhibition even in the cells that are inside the extracellular polymeric substance (EPS). Recent advances in designing new PSs to increase the production of ROS and the combination of aPDT with other therapies, especially pulsed electric fields (PEF), have contributed to enhanced biofilm inhibition. The PEF has proven to have antimicrobial effect once it is known that extensive chemical reactions occur when electric fields are applied. This type of treatment kills microorganisms not only due to membrane rupture but also due to the formation of reactive compounds including free oxygen, hydrogen, hydroxyl and hydroperoxyl radicals. So, this review aims to show the progress of aPDT and PEF against the biofilms, suggesting that the association of both methods can potentiate their effects and overcome biofilm infections.},
}

@article {pmid34495971,
year = {2021},
author = {Santos, AAN and Ribeiro, PDS and da França, GV and Souza, FN and Ramos, EAG and Figueira, CP and Reis, MG and Costa, F and Ristow, P},
title = {Leptospira interrogans biofilm formation in Rattus norvegicus (Norway rats) natural reservoirs.},
journal = {PLoS neglected tropical diseases},
volume = {15},
number = {9},
pages = {e0009736},
pmid = {34495971},
issn = {1935-2735},
support = {/WT_/Wellcome Trust/United Kingdom ; R01 AI052473/AI/NIAID NIH HHS/United States ; R01 TW009504/TW/FIC NIH HHS/United States ; R25 TW009338/TW/FIC NIH HHS/United States ; },
abstract = {Rattus norvegicus (Norway rat) is the main reservoir host of pathogenic Leptospira, the causative agent of leptospirosis, in urban environments. Pathogenic Leptospira forms biofilms in the environment, possibly contributing for bacterial survival and maintenance. Nonetheless, biofilms have not yet been studied in natural animal reservoirs presenting leptospiral renal carriage. Here, we described biofilm formation by pathogenic Leptospira inside the renal tubules of R. norvegicus naturally infected and captured in an urban slum endemic for leptospirosis. From the 65 rats carrying Leptospira in their kidneys, 24 (37%) presented biofilms inside the renal tubules. The intensity of leptospiral colonization in the renal tubules (OR: 1.00; 95% CI 1.05-1.1) and the type of occlusion pattern of the colonized renal tubules (OR: 3.46; 95% CI 1.20-9.98) were independently associated with the presence of Leptospira biofilm. Our data showed that Leptospira interrogans produce biofilms during renal chronic colonization in rat reservoirs, suggesting a possible role for leptospiral biofilms in the pathogenesis of leptospirosis and bacterial carriage in host reservoirs.},
}

@article {pmid34495103,
year = {2021},
author = {Viana, CS and Maske, TT and Signori, C and VAN DE Sande, FH and Oliveira, EF and Cenci, MS},
title = {Influence of caries activity and number of saliva donors: mineral and microbiological responses in a microcosm biofilm model.},
journal = {Journal of applied oral science : revista FOB},
volume = {29},
number = {},
pages = {e20200778},
pmid = {34495103},
issn = {1678-7765},
mesh = {Biofilms ; *Dental Caries ; Dental Caries Susceptibility ; Humans ; Minerals ; *Saliva ; Streptococcus mutans ; },
abstract = {OBJECTIVE: this study evaluated the mineral and microbiological response of biofilms originating from different types of saliva inoculum with distinct levels of caries activity.

METHODOLOGY: the biofilms grown over enamel specimens originated from saliva collected from a single donor or five donors with two distinct levels of caries activity (caries-active and caries-free) or from pooling saliva from ten donors (five caries-active and five caries-free). The percentage surface hardness change (%SHC) and microbiological counts served as outcome variables.

RESULTS: the caries activity of donors did not affect the %SHC values. Inoculum from five donors compared to a single donor showed higher %SHC values (p=0.019). Higher lactobacilli counts were observed when saliva from caries-active donors was used as the inoculum (p=0.017). Pooled saliva from both caries activity levels showed higher mutans streptococci counts (p<0.017).

CONCLUSION: Overall, pooled saliva increased the mineral response of the derived biofilms, but all the inoculum conditions formed cariogenic biofilms and caries lesions independently of caries activity.},
}

Biofilms
*Dental Caries
Dental Caries Susceptibility
Humans
Minerals
*Saliva
Streptococcus mutans

@article {pmid34494311,
year = {2021},
author = {Togo, Y},
title = {Editorial Comment to Intraluminal diamond-like carbon coating with anti-adhesion and anti-biofilm effects for uropathogens: A novel technology applicable to urinary catheters.},
journal = {International journal of urology : official journal of the Japanese Urological Association},
volume = {},
number = {},
pages = {},
doi = {10.1111/iju.14691},
pmid = {34494311},
issn = {1442-2042},
}

@article {pmid34494127,
year = {2021},
author = {Liu, Y and Luan, J},
title = {Letter to the Editor: Proper Skin Management in Breast Augmentation with a Periareolar Incision Prevents Implant Contamination and Biofilm-Related Capsular Contracture.},
journal = {Aesthetic plastic surgery},
volume = {},
number = {},
pages = {},
pmid = {34494127},
issn = {1432-5241},
}

@article {pmid34492866,
year = {2021},
author = {Zhou, B and Hou, P and Xiao, Y and Song, P and Xie, E and Li, Y},
title = {Visualizing, quantifying, and controlling local hydrodynamic effects on biofilm accumulation in complex flow paths.},
journal = {Journal of hazardous materials},
volume = {416},
number = {},
pages = {125937},
doi = {10.1016/j.jhazmat.2021.125937},
pmid = {34492866},
issn = {1873-3336},
mesh = {Biofilms ; Computer Simulation ; *Hydrodynamics ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; },
abstract = {Complex flow paths (CFPs) are commonly applied in precision equipment to accurately supply controllable fluids with designed structures. However, the presence of biofilms in CFPs causes quite a few unwanted issues, such as bio-erosion, clogging, or even health risks. To date, visualizing and quantifying the interaction between biofilm distribution and local hydrodynamics remains difficult, and the mechanism during the process is unclear. In this paper, the remodeling simulation method (3D industrial computed tomography scanning-inverse modeling-numerical simulation) and 16S rRNA high-throughput sequencing were integrated. The results indicated that local hydrodynamic characteristics significantly affected biofilm thicknesses on CFP surfaces (relative differences of 41.3-71.2%), which inversely influenced the local turbulence intensity. The average biofilm thicknesses exhibited a significant quadratic correlation with the near-wall hydraulic shear forces (r > 0.72, p < 0.05), and the biofilm reached a maximum thickness at 0.36-0.45 Pa. On the other hand, the near-wall hydraulic shear forces not only affected microbial community characteristics of biofilms, but they also influenced the number of microorganisms involved, which determined the biofilm accumulation thereafter. The PHYLUM Firmicutes and Proteobacteria were the dominant bacteria during the process. The results obtained in this paper could provide practical conceptions for the targeted control of biofilms and put forward more efficient controlling methods in commonly applied CFP systems.},
}

Biofilms
Computer Simulation
*Hydrodynamics
*Microbiota
RNA, Ribosomal, 16S/genetics

@article {pmid34492791,
year = {2021},
author = {Yang, T and Jiang, L and Cheng, L and Zheng, X and Bi, X and Wang, X and Zhou, X},
title = {Characteristics of size-segregated aerosols emitted from an aerobic moving bed biofilm reactor at a full-scale wastewater treatment plant.},
journal = {Journal of hazardous materials},
volume = {416},
number = {},
pages = {125833},
doi = {10.1016/j.jhazmat.2021.125833},
pmid = {34492791},
issn = {1873-3336},
mesh = {Aerosols/analysis ; *Biofilms ; Bioreactors ; Particulate Matter ; *Water Purification ; },
abstract = {Aerosol emissions from wastewater treatment plants (WWTPs) have been associated with health reverberation but studies about characteristics of size-segregated aerosol particulate matter (PM) are scarce. In this study, the measurement of particulate number size distribution in the range of < 10 µm, and the collection of PM10-2.5, PM2.5-1.0 and PM1.0, were conducted from an aerobic moving bed biofilm reactor (MBBR) at a full-scale WWTP. MBBR aerosols showed a unimodal number size distribution, with the majority of particles (>94%) in the ultrafine size range (<100 nm). For toxic metal(loid)s or potential pathogens, significant differences were found within MBBR aerosols (PM10-2.5, PM2.5-1.0, and PM1.0), and also between MBBR aerosols and wastewater. Both wastewater and ambient air had important source contributions for MBBR aerosols. The compositions of toxic metal(loid)s in PM1.0, and the populations of potential bacterial or fungal pathogens in PM10-2.5 and PM2.5-1.0, were dominated by that from wastewater. Compared to PM10-2.5 and PM2.5-1.0, PM1.0 had the highest aerosolization potential for the toxic metal(loid)s of As, Cd, Co, Cr, Li, Mn, Ni, U, and Zn, and the genera of Acinetobacter, Pseudomonas and Fusarium. Due to the size-segregated specialty, targeted measures should be employed to reduce the health risks. CAPSULE: The compositions of toxic metal(loid)s in PM1.0, and the populations of potential pathogens in PM10-2.5 and PM2.5-1.0, were dominated by that from wastewater.},
}

Aerosols/analysis
*Biofilms
Bioreactors
Particulate Matter
*Water Purification

@article {pmid34489569,
year = {2021},
author = {Deng, X and Zhang, C and Chen, J and Shi, Y and Ma, X and Wang, Y and Wang, Z and Yu, Z and Zheng, J and Chen, Z},
title = {Antibacterial and anti-biofilm activities of histidine kinase YycG inhibitors against Streptococcus agalactiae.},
journal = {The Journal of antibiotics},
volume = {},
number = {},
pages = {},
pmid = {34489569},
issn = {1881-1469},
support = {JCYJ20180302144345028//Shenzhen Science and Technology Innovation Commission/ ; JCYJ20180508162403996JCYJ20190809151817062;//Shenzhen Science and Technology Innovation Commission/ ; 2020A1515111146//Natural Science Foundation of Guangdong Province (Guangdong Natural Science Foundation)/ ; },
abstract = {This study aims to investigate the antibacterial and anti-biofilm activities of YycG inhibitors H2-60 and H2-81 against Streptococcus agalactiae. A total of 118 nonduplicate S. agalactiae clinical isolates were collected, and the minimal inhibitory concentrations (MICs) of H2-60 and H2-81 were determined. H2-60 and H2-81 inhibit biofilm formation of S. agalactiae were detected by crystal violet staining, and against established biofilms of S. agalactiae were observed by confocal laser scanning microscope. Inhibitory effect of H2-60 and H2-81 on the phosphorylation activity of the HisKA domain of YycG' protein was measured. The MIC50/MIC90 was 3.13/6.25 μM for H2-60 and 6.25/12.5 μM for H2-81 against S. agalactiae, respectively. S. agalactiae planktonic cells can be decreased by H2-60 or H2-81 for more than 3 × log10 CFU ml-1 after 24 h treatment. Biofilm formation of 8 S. agalactiae strains (strong biofilm producers) was significantly reduced after treated with 1/4 × MIC of H2-60 or H2-81 for 24 h. H2-60 and H2-81 could reduce 45.79% and 29.56% of the adherent cells in the established biofilm of S. agalactiae after 72 h treatment, respectively. H2-60 combined with daptomycin reduced 83.63% of the adherent cells in the established biofilm of S. agalactiae, which was significantly better than that of H2-60 (45.79%) or daptomycin (55.07%) alone. The half maximal inhibitory concentrations (IC50) were 35.6 μM for H2-60 and 46.3 μM for H2-81 against the HisKA domain of YycG' protein. In conclusion, YycG inhibitors H2-60 and H2-81 exhibit excellent antibacterial and anti-biofilm activities against S. agalactiae.},
}

@article {pmid34489479,
year = {2021},
author = {Duraj-Thatte, AM and Praveschotinunt, P and Nash, TR and Ward, FR and Nguyen, PQ and Joshi, NS},
title = {Author Correction: Modulating bacterial and gut mucosal interactions with engineered biofilm matrix proteins.},
journal = {Scientific reports},
volume = {11},
number = {1},
pages = {18033},
doi = {10.1038/s41598-021-96036-w},
pmid = {34489479},
issn = {2045-2322},
}

@article {pmid34488149,
year = {2021},
author = {Dong, X and Zhu, L and Jiang, P and Wang, X and Liu, K and Li, C and Li, D},
title = {Seasonal biofilm formation on floating microplastics in coastal waters of intensified marinculture area.},
journal = {Marine pollution bulletin},
volume = {171},
number = {},
pages = {112914},
doi = {10.1016/j.marpolbul.2021.112914},
pmid = {34488149},
issn = {1879-3363},
abstract = {The environmental pollution caused by microplastics has received increasing attention recently. In this paper, we present the results of research into the bacterium attached to microplastics in four coastal mariculture zones in southeast China during winter and summer. Polyethene and polypropylene are the main microplastics in the surface water of mariculture area. The differences between the bacteria species composition found on the surface of microplastics in winter and summer were less than that found in the planktonic bacteria, indicating that biofilms protect the bacterium that live inside. Potentially pathogenic Vibrio and Pseudomonas spp. were more abundant in samples from ShanTou and QuanZhou during the summer. Bacteria related to the degradation of microplastics were found extensively on the surface of microplastics at all of the sampling sites. More attention should be paid to the risks resulting from the accumulation of harmful bacteria on microplastic surfaces during the summer.},
}

@article {pmid34486975,
year = {2021},
author = {Kalamara, M and Abbott, JC and MacPhee, CE and Stanley-Wall, NR},
title = {Biofilm hydrophobicity in environmental isolates of Bacillus subtilis.},
journal = {Microbiology (Reading, England)},
volume = {167},
number = {9},
pages = {},
doi = {10.1099/mic.0.001082},
pmid = {34486975},
issn = {1465-2080},
support = {BB/P001335/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/R012415/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/M010996/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; },
abstract = {Biofilms are communities of bacteria that are attached to a surface and surrounded by an extracellular matrix. The extracellular matrix protects the community from stressors in the environment, making biofilms robust. The Gram-positive soil bacterium Bacillus subtilis, particularly the isolate NCIB 3610, is widely used as a model for studying biofilm formation. B. subtilis NCIB 3610 forms colony biofilms that are architecturally complex and highly hydrophobic. The hydrophobicity is linked, in part, to the localisation of the protein BslA at the surface of the biofilm, which provides the community with increased resistance to biocides. As most of our knowledge about B. subtilis biofilm formation comes from one isolate, it is unclear if biofilm hydrophobicity is a widely distributed feature of the species. To address this knowledge gap, we collated a library of B. subtilis soil isolates and acquired their whole genome sequences. We used our novel isolates to examine biofilm hydrophobicity and found that, although BslA is encoded and produced by all isolates in our collection, hydrophobicity is not a universal feature of B. subtilis colony biofilms. To test whether the matrix exopolymer poly γ-glutamic acid could be masking hydrophobicity in our hydrophilic isolates, we constructed deletion mutants and found, contrary to our hypothesis, that the presence of poly γ-glutamic acid was not the reason for the observed hydrophilicity. This study highlights the natural variation in the properties of biofilms formed by different isolates and the importance of using a more diverse range of isolates as representatives of a species.},
}

@article {pmid34485498,
year = {2021},
author = {Boswell, MT and Cockeran, R},
title = {Effect of antimicrobial peptides on planktonic growth, biofilm formation and biofilm-derived bacterial viability of Streptococcus pneumoniae.},
journal = {Southern African journal of infectious diseases},
volume = {36},
number = {1},
pages = {226},
pmid = {34485498},
issn = {2313-1810},
abstract = {Streptococcus pneumoniae is a leading cause of pneumonia mortality globally. Pneumococcal disease is often associated with prolonged colonisation of hosts and this process is facilitated by biofilm formation that is largely resistant to conventional antibiotics. We investigated the effects of antimicrobial peptides (AMPs) lysozyme, lactoferrin, LL37 and a combination of all three on planktonic growth, biofilm formation and biofilm-derived bacterial viability by S. pneumoniae, serotype 23F. Planktonic growth and biofilm-derived bacterial viability were determined using standard colony-forming techniques, while biofilm formation was measured using a crystal violet based spectrophotometric method. Relative to controls, lysozyme significantly reduced biofilm formation (0.08 OD vs. 0.10 OD at 570 nm, p = 0.01), while LL37 and the AMP combination increased biofilm formation (0.14 OD vs. 0.10 OD at 570 nm, p = 0.01). The combination of AMPs significantly decreased planktonic growth (1.10 × 108 colony-forming units per millilitres [CFU/mL] vs. 2.13 × 108 CFU/mL, p = 0.02). Biofilm-derived bacterial viability was greatly reduced by exposure to a combination of AMPs (1.05 × 105 CFU/mL vs. 1.12 × 106 CFU/mL, p = 3.60 × 10-8). Streptococcus pneumoniae displays marked resistance to the individual AMPs. A combination of lysozyme, lactoferrin and LL37 effectively inhibited planktonic growth and biofilm-derived bacterial viability; however, persister cell growth was still evident after exposure.},
}

@article {pmid34485167,
year = {2021},
author = {Ramírez-Granillo, A and Bautista-Hernández, LA and Bautista-De Lucío, VM and Magaña-Guerrero, FS and Domínguez-López, A and Córdova-Alcántara, IM and Pérez, NO and Martínez-Rivera, MLA and Rodríguez-Tovar, AV},
title = {Microbial Warfare on Three Fronts: Mixed Biofilm of Aspergillus fumigatus and Staphylococcus aureus on Primary Cultures of Human Limbo-Corneal Fibroblasts.},
journal = {Frontiers in cellular and infection microbiology},
volume = {11},
number = {},
pages = {646054},
pmid = {34485167},
issn = {2235-2988},
abstract = {Background: Coinfections with fungi and bacteria in ocular pathologies are increasing at an alarming rate. Two of the main etiologic agents of infections on the corneal surface, such as Aspergillus fumigatus and Staphylococcus aureus, can form a biofilm. However, mixed fungal-bacterial biofilms are rarely reported in ocular infections. The implementation of cell cultures as a study model related to biofilm microbial keratitis will allow understanding the pathogenesis in the cornea. The cornea maintains a pathogen-free ocular surface in which human limbo-corneal fibroblast cells are part of its cell regeneration process. There are no reports of biofilm formation assays on limbo-corneal fibroblasts, as well as their behavior with a polymicrobial infection.

Objective: To determine the capacity of biofilm formation during this fungal-bacterial interaction on primary limbo-corneal fibroblast monolayers.

Results: The biofilm on the limbo-corneal fibroblast culture was analyzed by assessing biomass production and determining metabolic activity. Furthermore, the mixed biofilm effect on this cell culture was observed with several microscopy techniques. The single and mixed biofilm was higher on the limbo-corneal fibroblast monolayer than on abiotic surfaces. The A. fumigatus biofilm on the human limbo-corneal fibroblast culture showed a considerable decrease compared to the S. aureus biofilm on the limbo-corneal fibroblast monolayer. Moreover, the mixed biofilm had a lower density than that of the single biofilm. Antibiosis between A. fumigatus and S. aureus persisted during the challenge to limbo-corneal fibroblasts, but it seems that the fungus was more effectively inhibited.

Conclusion: This is the first report of mixed fungal-bacterial biofilm production and morphological characterization on the limbo-corneal fibroblast monolayer. Three antibiosis behaviors were observed between fungi, bacteria, and limbo-corneal fibroblasts. The mycophagy effect over A. fumigatus by S. aureus was exacerbated on the limbo-corneal fibroblast monolayer. During fungal-bacterial interactions, it appears that limbo-corneal fibroblasts showed some phagocytic activity, demonstrating tripartite relationships during coinfection.},
}

@article {pmid34485075,
year = {2021},
author = {Svensson Malchau, K and Tillander, J and Zaborowska, M and Hoffman, M and Lasa, I and Thomsen, P and Malchau, H and Rolfson, O and Trobos, M},
title = {Biofilm properties in relation to treatment outcome in patients with first-time periprosthetic hip or knee joint infection.},
journal = {Journal of orthopaedic translation},
volume = {30},
number = {},
pages = {31-40},
pmid = {34485075},
issn = {2214-031X},
abstract = {Background: Periprosthetic joint infections (PJI) are challenging complications following arthroplasty. Staphylococci are a frequent cause of PJI and known biofilm producers. Biofilm formation decreases antimicrobial susceptibility, thereby challenging favourable treatment outcomes. The aims of this study were to characterize the biofilm abilities and antimicrobial susceptibilities of staphylococci causing first-time PJI and correlate them to clinical outcome (infection resolution and recurrence).

Methods: Reoperations for PJI of the hip or knee between 1st January 2012 to 30th June 2015 performed at the Sahlgrenska University Hospital were identified in a local database. Medical records were reviewed and clinical parameters recorded for patients whose intraoperative bacterial isolates had been stored at the clinical laboratory. Staphylococcal strains isolated from reoperations due to first-time PJI were characterised by their ability to form biofilms using the microtiter plate test. Antimicrobial susceptibility of the strains was determined by minimum inhibitory concentration (MIC) when grown planktonically, and by minimum biofilm eradication concentration (MBEC) when grown as biofilms. MBEC determination was conducted using the Calgary biofilm device (CBD) and a custom-made antimicrobial susceptibility plate containing eight clinically relevant antimicrobial agents.

Results: The study group included 49 patients (70 bacterial strains) from first-time PJI, whereof 24 (49%) patients had recurrent infection. Strong biofilm production was significantly associated with recurrent infection. Patients infected with strong biofilm producers had a five-fold increased risk for recurrent infection. Strains grown as biofilms were over 8000 times more resistant to antimicrobial agents compared to planktonic cultures. Biofilms were more susceptible to rifampicin compared to other antimicrobials in the assay. Increased biofilm susceptibility (MBEC ​> ​MIC) was observed for the majority of the bacterial strains and antimicrobial agents.

Conclusions: Strong biofilm production was significantly associated with increased antimicrobial resistance and PJI recurrence. This underscores the importance of determining biofilm production and susceptibility as part of routine diagnostics in PJI. Strong staphylococcal biofilm production may have implications on therapeutic choices and suggest more extensive surgery. Furthermore, despite the increased biofilm resistance to rifampicin, results from this study support its use in staphylococcal PJI.

Like for many biomaterial-associated infections, staphylococci are a common cause of PJI. Their ability to adhere to surfaces and produce biofilms on medical devices is proposed to play a role. However, clinical studies where biofilm properties are directly linked to patient outcome are scarce. This study demonstrates that the majority of staphylococci isolated from first-time PJI were biofilm producers with increased antimicrobial resistance. Patients suffering an infection caused by a staphylococcal strain with strong biofilm production ability had a five-fold greater risk of recurrent infection. This novel finding suggests the importance of evaluating biofilm production as a diagnostic procedure for the guidance of treatment decisions in PJI.},
}

@article {pmid34484613,
year = {2021},
author = {Liao, MH and Wang, XR and Hsu, WL and Tzen, JTC},
title = {Pu'er tea rich in strictinin and catechins prevents biofilm formation of two cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus.},
journal = {Journal of dental sciences},
volume = {16},
number = {4},
pages = {1331-1334},
pmid = {34484613},
issn = {2213-8862},
abstract = {Cariogenic bacteria, such as Streptococcus mutans and Streptococcus sobrinus, are main pathogens responsible for human dental caries. Pu'er tea is empirically observed to prevent tooth decay. Besides caffeine and catechins commonly found in oolong tea, strictinin is also found as an abundant phenolic compound in Pu'er tea. Infusion of Pu'er tea as well as single compound, strictinin, caffeine or (-)-epigallocatechin gallate (EGCG) was examined for its inhibitory effects on S. mutans and S. sobrinus. Relatively weak inhibition of bacterial growth was observed for these Pu'er tea constituents. However, biofilm formation of S. mutans or S. sobrinus was strongly prevented by the infusion of Pu'er tea as well as by strictinin or EGCG, but not caffeine. Relatively, strictinin showed a higher potency than EGCG to prevent biofilm formation. Anti-caries effect of Pu'er tea seems to be resulted from the prevention of biofilm formation of cariogenic bacteria mainly by strictinin and catechins.},
}

@article {pmid34482605,
year = {2021},
author = {Huang, Z and Dai, H and Zhang, X and Wang, Q and Sun, J and Deng, Y and Shi, P},
title = {BSC2 induces multidrug resistance via contributing to the formation of biofilm in Saccharomyces cerevisiae.},
journal = {Cellular microbiology},
volume = {},
number = {},
pages = {e13391},
doi = {10.1111/cmi.13391},
pmid = {34482605},
issn = {1462-5822},
support = {2232021G-04//Fundamental Research Funds for the Central Universities/ ; 2021-ZJ-Y14//Qinghai Provincial Key Laboratory of Qinghai-Tibet Plateau Biological Resources/ ; },
abstract = {Biofilm plays an important role in fungal multidrug resistance (MDR). Our previous studies showed that BSC2 is involved in resistance to amphotericin B (AMB) through antioxidation in Saccharomyces cerevisiae. In this study, the overexpression of BSC2 and IRC23 induced strong MDR in S. cerevisiae. BSC2-overexpression affected cellular flocculation, cell surface hydrophobicity, biofilm formation and invasive growth. However, it failed to induce caspofungin (CAS) resistance and affect the invasive growth in FLO mutant strains (FLO11Δ, FLO1Δ, FLO8Δ and TUP1Δ). Furthermore, the overexpression of BSC2 compensated for chitin synthesis defects to maintain the cell wall integrity and significantly reduced the cell morphology abnormality induced by CAS. However, it could not repair the cell wall damage caused by CAS in the FLO mutant strains. Although BSC2 overexpression increased the level of mannose in the cell wall, DPM1 overexpression in both BY4741 and bsc2∆ could confer resistance to CAS and AMB. In addition, BSC2 overexpression significantly increased the mRNA expression of FLO11, FLO1, FLO8 and TUP1. BSC2 may function as a regulator of FLO genes and be involved in cell wall integrity in yeast. Taken together, our data demonstrate that BSC2 induces MDR in a FLO pathway-dependent manner via contributing to the formation of biofilms in S. cerevisiae. TAKE AWAYS: Overexpression of BSC2 induced strong MDR in S. cerevisiae. BSC2 affected cellular flocculation, CSH, biofilm formation and invasive growth. BSC2 could not repair the cell wall damage caused by CAS in the FLO mutants. BSC2 may function as a regulator of FLO genes to maintain cell wall integrity. BSC2 promotes biofilm formation in a FLO pathway-dependent manner to induce MDR.},
}

@article {pmid34482564,
year = {2021},
author = {Watari, S and Wada, K and Araki, M and Sadahira, T and Ousaka, D and Oozawa, S and Nakatani, T and Imai, Y and Kato, J and Kariyama, R and Watanabe, T and Nasu, Y},
title = {Intraluminal diamond-like carbon coating with anti-adhesion and anti-biofilm effects for uropathogens: A novel technology applicable to urinary catheters.},
journal = {International journal of urology : official journal of the Japanese Urological Association},
volume = {},
number = {},
pages = {},
doi = {10.1111/iju.14675},
pmid = {34482564},
issn = {1442-2042},
support = {19K18585//Grant-in-Aid for Early Career Scientists/ ; //Ministry of Education, Culture, Sports, Science and Technology, Japan/ ; },
abstract = {OBJECTIVES: To examine anti-adhesion and anti-biofilm effects of a diamond-like carbon coating deposited via a novel technique on the inner surface of a thin silicon tube.

METHODS: Diamond-like carbon coatings were deposited into the lumen of a silicon tube with inner diameters of 2 mm. The surface of the diamond-like carbon was evaluated using physicochemical methods. We used three clinical isolates including green fluorescent protein-expressing Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus. We employed a continuous flow system for evaluation of both bacterial adhesion and biofilm formation. Bacterial adhesion assays consisted of counting the number of colony-forming units and visualization of adhered bacterial cells by scanning electron microscope to evaluate the diamond-like carbon-coated/uncoated samples. The biofilm structure was analyzed by confocal laser scanning microscopy on days 3, 5, 7 and 14 for green fluorescent protein-expressing Pseudomonas aeruginosa.

RESULTS: The smooth and carbon-rich structure of the intraluminal diamond-like carbon film remained unchanged after the experiments. The numbers of colony-forming units suggested lower adherence of green fluorescent protein-expressing Pseudomonas aeruginosa and Escherichia coli in the diamond-like carbon-coated samples compared with the uncoated samples. The scanning electron microscope images showed adhered green fluorescent protein-expressing Pseudomonas aeruginosa cells without formation of microcolonies on the diamond-like carbon-coated samples. Finally, biofilm formation on the diamond-like carbon-coated samples was lower until at least day 14 compared with the uncoated samples.

CONCLUSIONS: Intraluminal diamond-like carbon coating on a silicone tube has anti-adhesion and anti-biofilm effects. This technology can be applied to urinary catheters made from various materials.},
}

@article {pmid34481121,
year = {2021},
author = {Eze, EC and El Zowalaty, ME and Pillay, M},
title = {Antibiotic resistance and biofilm formation of Acinetobacter baumannii isolated from high-risk effluent water in tertiary hospitals in South Africa.},
journal = {Journal of global antimicrobial resistance},
volume = {27},
number = {},
pages = {82-90},
doi = {10.1016/j.jgar.2021.08.004},
pmid = {34481121},
issn = {2213-7173},
abstract = {OBJECTIVES: Discharge of drug-resistant, biofilm-forming pathogens from hospital effluent water into municipal wastewater treatment plants poses a public health concern. This study examined the relationship between antibiotic resistance levels and biofilm formation of Acinetobacter baumannii strains isolated from hospital effluents.

METHODS: Antibiotic susceptibility of 71 A. baumannii isolates was evaluated by the Kirby-Bauer disk diffusion method. Minimum inhibitory concentrations (MICs) were determined by the agar dilution method, while the minimum biofilm eradication concentration (MBEC) was determined by the broth dilution method. Genotyping was performed for plasmid DNA. Biofilm formation was evaluated by the microtitre plate method and was quantified using crystal violet. A P-value of <0.05 was regarded as statistically significant in all tests.

RESULTS: Extensively drug-resistant (XDR) strains made up 58% of the isolates, while multidrug-resistant (MDR) and pandrug-resistant (PDR) strains made up 50% of the isolates from final effluent. The MBEC of ciprofloxacin increased by 255-fold, while that of ceftazidime was as high as 63-1310-fold compared with their respective MICs. Isolates were classified into four plasmid pattern groups with no association between biofilm formation and plasmid type (P = 0.0921). The degree of biofilm formation was independent of the level of antibiotic resistance, although MDR, XDR and PDR isolates produced significant biofilm biomass (P = 0.2580).

CONCLUSION: These results suggest that hospital effluent is a potential source of MDR biofilm-forming A. baumannii strains. Appropriate treatment and disposal of effluents are essential to prevent the presence of drug-resistant pathogens in wastewater.},
}

@article {pmid34480246,
year = {2021},
author = {Joshi, K and Navalgund, L and Rathod, K and Shet, VB and Srinikethan, G and Shet, T and Rachitha, U and Anand, A},
title = {Biofilm characterization in removal of total chemical oxygen demand and nitrate from wastewater using draft tube spouted bed reactor.},
journal = {Biotechnology letters},
volume = {43},
number = {10},
pages = {2001-2009},
pmid = {34480246},
issn = {1573-6776},
abstract = {The present paper investigates the effect of dilution rate on the removal of total chemical oxygen demand and nitrate in the draft tube spouted bed reactor and morphological characteristics of biofilms formed by microorganisms of mixed culture on granular activated carbon (GAC). The nitrate and total chemical oxygen demand (COD) decreased from 97 to 81% and 95% to 87% respectively with increase in dilution rate from 0.6/h to 1.5/h showing that residence time in the reactor governs the nitrate and total COD reduction efficiency. Lower dilution rates favor higher production of biomass and extracellular polymeric substances (EPS). It was observed that the nitrate and total COD reduction rate increased with time along with simultaneous increase in EPS production. Thus, the performance of a reactor in terms of dynamic and steady-state biofilm characteristics associated with nitrate and organic reduction is a strong function of dilution rate. Hence these findings indicate that a draft tube spouted bed reactor is capable of simultaneously reducing total organics and nitrogen in industrial/municipal wastewater, as this reactor possesses two distinct regions aerobic and anoxic conditions which can prevail in different parts of a reactor.},
}

@article {pmid34479647,
year = {2021},
author = {Toledo-Silva, B and de Souza, FN and Mertens, K and Piepers, S and Haesebrouck, F and De Vliegher, S},
title = {Bovine-associated non-aureus staphylococci suppress Staphylococcus aureus biofilm dispersal in vitro yet not through agr regulation.},
journal = {Veterinary research},
volume = {52},
number = {1},
pages = {114},
pmid = {34479647},
issn = {1297-9716},
support = {G0H2516N (AUGE/15/05)//fonds wetenschappelijk onderzoek/ ; },
abstract = {Biofilm formation is a significant virulence factor in Staphylococcus (S.) aureus strains causing subclinical mastitis in dairy cows. A role of environmental signals and communication systems in biofilm development, such as the agr system in S. aureus, is suggested. In the context of multispecies biofilm communities, the presence of non-aureus staphylococci (NAS) might influence S. aureus colonization of the bovine mammary gland, yet, such interspecies interactions have been poorly studied. We determined whether 34 S. chromogenes, 11 S. epidermidis, and 14 S. simulans isolates originating from bovine milk samples and teat apices (TA) were able to affect biofilm formation and dispersion of S. aureus, and if so, how isolate traits such as the capacity to regulate the S. aureus agr quorum sensing system are determinants in this process. The capacity of an agr-positive S. aureus strain to form biofilm was increased more in the presence of S. chromogenes than in the presence of S. simulans and S. epidermidis isolates and in the presence of NAS isolates that do not harbor biofilm related genes. On the other hand, biofilm dispersion of this particular S. aureus strain was suppressed by NAS as a group, an effect that was more pronounced by isolates from TA. Furthermore, the observed effects on biofilm formation and dispersion of the agr-positive S. aureus strain as well as of an agr-negative S. aureus strain did not depend on the capacity of NAS to repress the agr system.},
}

@article {pmid34478899,
year = {2021},
author = {Ghazi, M and Pourhajibagher, M and Bahador, A and Chiniforush, N and Dadpour, S and Dadpour, Y},
title = {Evaluation of adding nanosized natural zeolite to photodynamic therapy against P. gingivalis biofilm on titanium disks.},
journal = {Photodiagnosis and photodynamic therapy},
volume = {36},
number = {},
pages = {102519},
doi = {10.1016/j.pdpdt.2021.102519},
pmid = {34478899},
issn = {1873-1597},
abstract = {BACKGROUND: Antibacterial photodynamic therapy (aPDT) can be used as an adjunctive therapy for eliminating bacterial biofilm. The application of nanotechnology in aPDT, which is a growing trend, has improved the delivery of photosensitizers (PSs) into microorganisms. Encapsulation of molecules and ions is considered an outstanding potential feature of zeolites. This study sought to enhance the effect of aPDT using a diode laser (810 nm) with a potential PS, indocyanine green (ICG), combined with nanosized natural zeolite (NZ), against biofilm of P. gingivalis on sandblasted, large-grit, and acid-etched (SLA) implant titanium disks surface.

METHODS: A bacterial suspension of standard P. gingivalis (™ATCC® 33277) strains was prepared. To prepare bacterial biofilm, the titanium disks were added to 48 microtubes containing bacterial suspension, and divided into eight groups, i.e., the control groups (positive and negative), and 6 test groups (ICG; NZ; Diod laser; NZ+ICG; aPDT; NZ+aPDT). After the treatments, the total number of colony-forming units per disk was calculated. Finally, the data was analyzed, and the eight groups were compared together.

RESULTS: The highest reduction in the number of P. gingivalis was seen in group 8 (NZ+aPDT) with 3.55 log10 CFU/ml and the antibacterial effect of 45.7% compared with the negative control group. Conversley, group 5 (Diode Laser solely) represented the highest mean of colony count with the lowest antibacterial effects per disk (6.42 log10 CFU/ml, 1.8%).

CONCLUSIONS: The antibacterial effect of NZ+aPDT against P. gingivalis biofilm was noticeable. Thus, adding NZ to ICG improved the result of aPDT in this study.},
}

@article {pmid34475672,
year = {2021},
author = {Garg, A and Mala, K and Kamath, PM},
title = {Biofilm models in endodontics-A narrative review.},
journal = {Journal of conservative dentistry : JCD},
volume = {24},
number = {1},
pages = {2-9},
pmid = {34475672},
issn = {0972-0707},
abstract = {The knowledge of biofilm and its eradication from the root canal system are of utmost importance in the clinical practice of an endodontist. Various treatment strategies and protocols have been demonstrated and discussed by numerous clinicians and researchers, on these models, that play an important role in the treatment outcome . Once a biofilm model is developed by considering various factors, several methods can be used to assess the biofilms formed on these models. This review discusses the importance of biofilm models in endodontics, types of biofilm models and factors associated with developing and the methods to evaluate these models.},
}

@article {pmid34474869,
year = {2021},
author = {Song, X and Liu, P and Liu, X and Wang, Y and Wei, H and Zhang, J and Yu, L and Yan, X and He, Z},
title = {Dealing with MDR bacteria and biofilm in the post-antibiotic era: Application of antimicrobial peptides-based nano-formulation.},
journal = {Materials science & engineering. C, Materials for biological applications},
volume = {128},
number = {},
pages = {112318},
doi = {10.1016/j.msec.2021.112318},
pmid = {34474869},
issn = {1873-0191},
mesh = {*Anti-Bacterial Agents/pharmacology ; *Antimicrobial Cationic Peptides/pharmacology ; Bacteria ; Biofilms ; Pore Forming Cytotoxic Proteins ; },
abstract = {The rapid development of multidrug-resistant (MDR) bacteria due to the improper and overuse of antibiotics and the ineffective performance of antibiotics against the difficult-to-treat biofilm-related infections (BRIs) have urgently called for alternative antimicrobial agents and strategies in combating bacterial infections. Antimicrobial peptides (AMPs), owing to their compelling antimicrobial activity against MDR bacteria and BRIs without causing bacteria resistance, have attracted extensive attention in the research field. With the development of nanomaterial-based drug delivery strategies, AMPs-based nano-formulations have significantly improved the therapeutic effects of AMPs by ameliorating their hydrolytic stability, half-life in vivo, and solubility as well as reducing the cytotoxicity and hemolysis, etc. This review has comprehensively summarized the application AMPs-based nano-formulation in various bacterial infections models, including bloodstream infections (specifically sepsis), pulmonary infections, chronic wound infections, gastrointestinal infections, among others. The design of the nanomaterial-based drug delivery systems and the therapeutic effects of the AMPs-based nano-formulations in literature have been categorized and in details discussed. Overall, this review provides insights into the advantages and disadvantages of the current developed AMPs-based nano-formulations in literature for the treatment of bacterial infections, bringing inspirations and suggestions for their future design in the way towards clinical translation.},
}

*Anti-Bacterial Agents/pharmacology
*Antimicrobial Cationic Peptides/pharmacology
Bacteria
Biofilms
Pore Forming Cytotoxic Proteins

@article {pmid34474857,
year = {2021},
author = {Qiu, G and Wu, H and Huang, M and Ma, T and Schneider, A and Oates, TW and Weir, MD and Xu, HHK and Zhao, L},
title = {Novel calcium phosphate cement with biofilm-inhibition and platelet lysate delivery to enhance osteogenesis of encapsulated human periodontal ligament stem cells.},
journal = {Materials science & engineering. C, Materials for biological applications},
volume = {128},
number = {},
pages = {112306},
pmid = {34474857},
issn = {1873-0191},
support = {R21 DE029611/DE/NIDCR NIH HHS/United States ; },
mesh = {Biofilms ; Calcium Phosphates/pharmacology ; Cell Differentiation ; Cells, Cultured ; Humans ; *Osteogenesis ; *Periodontal Ligament ; Staphylococcus aureus ; Stem Cells ; },
abstract = {Osteomyelitis is caused by Staphylococcus aureus (S. aureus), with associated progressive bone loss. This study developed for the first time a calcium phosphate cement (CPC) for delivery of doxycycline (DOX) and human platelet lysate (hPL) to fight against S. aureus infection and enhance the osteogenesis of human periodontal ligament stem cells (hPDLSCs). Chitosan-containing CPC scaffolds were fabricated in the absence (CPCC) or presence of DOX (CPCC+DOX). In addition, hPL was encapsulated in alginate microbeads and incorporated into CPCC+DOX (CPCC+DOX+ hPL). Flexural strength of CPCC+DOX + hPL was (5.56 ± 0.55) MPa, lower than (8.26 ± 1.6) MPa of CPCC+DOX (p < 0.05), but exceeding the reported strength of cancellous bone. CPCC+DOX and CPCC+DOX + hPL exhibited strong antibacterial activity against S. aureus, reducing biofilm CFU by 4 orders of magnitude. The hPDLSCs encapsulated in microbeads were co-cultured with the CPCs. The hPDLSCs were able to be released from the microbeads and showed a high proliferation rate, increasing by about 8 folds at 14 days for all groups. The hPL was released from the scaffold and promoted the osteogenic differentiation of hPDLSCs. ALP activity was 28.07 ± 5.15 mU/mg for CPCC+DOX + hPL, higher than 17.36 ± 2.37 mU/mg and 1.34 ± 0.37 mU/mg of CPCC+DOX and CPCC, respectively (p < 0.05). At 7 days, osteogenic genes (ALP, RUNX2, COL-1, and OPN) in CPCC+DOX + hPL were 3-10 folds those of control. The amount of hPDLSC-synthesized bone mineral with CPCC+DOX + hPL was 3.8 folds that of CPCC (p < 0.05). In summary, the novel CPC + DOX + hPL-hPDLSCs scaffold exhibited strong antibacterial activity, excellent cytocompatibility and hPDLSC osteogenic differentiation, showing a promising approach for treatment and prevention of bone infection and enhancement of bone regeneration.},
}

Biofilms
Calcium Phosphates/pharmacology
Cell Differentiation
Cells, Cultured
Humans
*Osteogenesis
*Periodontal Ligament
Staphylococcus aureus
Stem Cells

@article {pmid34474848,
year = {2021},
author = {Mendhi, J and Ramachandra, SS and Prasadam, I and Ivanovski, S and Yang, Y and Xiao, Y},
title = {Endogenous nitric oxide-generating surfaces via polydopamine-copper coatings for preventing biofilm dispersal and promoting microbial killing.},
journal = {Materials science & engineering. C, Materials for biological applications},
volume = {128},
number = {},
pages = {112297},
doi = {10.1016/j.msec.2021.112297},
pmid = {34474848},
issn = {1873-0191},
mesh = {Animals ; Biofilms ; Coated Materials, Biocompatible/pharmacology ; *Copper/pharmacology ; Homicide ; Indoles ; *Nitric Oxide ; Polymers ; Sheep ; Surface Properties ; Titanium/pharmacology ; },
abstract = {INTRODUCTION: Peri-implantitis is a bacterially induced inflammatory disease which affects the hard and soft tissues around a dental implant. Microbial biofilm formation is an important causative factor in peri-implantitis. The aim of this study is to develop an effective multifunctional surface coating for antimicrobial property and to counteract oral biofilm-associated infections via a single polydopamine copper coating (PDAM@Cu) on titanium implant surface to regulate endogenous nitric oxide (NO) generation.

METHODS: PDAM@Cu coatings were made with different concentrations of CuCl2 on titanium surfaces with a simple dip coating technique. Coatings were characterised to evaluate Cu concentrations as well as NO release rates from the coatings. Further, salivary biofilms were made on the coatings using Brain Heart Infusion (BHI) media in an anaerobic chamber. Biofilms were prepared with three different mixtures, one of which was saliva only, the second had an addition of sheep's blood, and the third was prepared with NO donors S-nitrosoglutathione (GSNO) and L-glutathione (GSH) in the mixture of saliva and blood to evaluate the effects of endogenously produced NO on biofilms. The effectiveness of coated surfaces on biofilms were assessed using four different methods, namely, crystal violet assay, scanning electron microscopy imaging, 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) metabolic assay, and live/dead staining.

RESULTS: NO release rates could be controlled with different Cu concentration in PDAM@Cu coatings. NO generated from the PDAM@Cu coatings effectively induced dispersal of biofilms shown by the reduction in biofilm biomass as well as reduced biofilm attachment in samples prepared with blood and NO donors. Cu ions released from the PDAM@Cu coatings resulted in killing of the dispersed bacteria, which was evidenced by the live/dead cell staining and reduced metabolic activity noted from the XTT assay. In contrast, samples prepared with saliva showed no significant reduction in biofilms, indicating the important effect of endogenously generated NO on biofilm dispersal.

CONCLUSION: In conclusion, PDAM@Cu coatings with NO generating surfaces have a dual anti-biofilm function, with a synergistic effect on biofilm dispersal from regulated NO generation and bactericidal effects from Cu ions from the coatings.},
}

Animals
Biofilms
Coated Materials, Biocompatible/pharmacology
*Copper/pharmacology
Homicide
Indoles
*Nitric Oxide
Polymers
Sheep
Surface Properties
Titanium/pharmacology

@article {pmid34473354,
year = {2021},
author = {Leelapornpisid, W and Novak-Frazer, L and Qualtrough, A and Rautemaa-Richardson, R},
title = {Effectiveness of D,L-2-hydroxyisocaproic acid (HICA) and alpha-mangostin against endodontopathogenic microorganisms in a multispecies bacterial-fungal biofilm in an ex vivo tooth model.},
journal = {International endodontic journal},
volume = {},
number = {},
pages = {},
doi = {10.1111/iej.13623},
pmid = {34473354},
issn = {1365-2591},
abstract = {AIM: To develop a defined multispecies root canal biofilm model ex vivo, and to perform viable compositional analysis following D,L-2-hydroxyisocaproic acid (HICA), alpha-mangostin, Calcicur® , and Odontopaste® exposure.

METHODOLOGY: Time-kill assays were conducted in vitro using HICA, alpha-mangostin, Calcicur® , Odontopaste® , and saline solution on the planktonic cultures of C. albicans, E. faecalis, L. rhamnosus, and S. gordonii. Human root dentine blocks were prepared (n = 100) ex vivo, and multispecies suspensions containing each of 1.5 × 108 CFU/mL C. albicans, E. faecalis, L. rhamnosus, and S. gordonii in brain heart infusion (BHI) were incubated within the root canals for 21 days. Canals (n = 20/group) were then exposed to medicaments for 7 days. Samples taken from the inner (first 0.1 mm) and deeper (second 0.1 mm) dentine by drilling with Ash Steel Burs No. 5 and No. 6, and residual roots were cultured in broth for 24 h. Cell growth was detected by spectrophotometry and confirmed by culture on agar. The other set of inner dentine, deeper dentine, and residual root samples were sonicated, and then exposed with 50 μM PMA before DNA was extracted using the QIAamp DNA mini kit. Real-time quantitative PCR was performed to determine the biofilm composition as well as the number of live and total cells remaining in the biofilm following each treatment. The OD data were analysed with Kruskal-Wallis and Friedman with Wilcoxon signed-rank test between and within groups, respectively, agar culture and qPCR data with Pearson chi-square with Mann-Whitney and Cochran with McNemar tests, respectively (p < .0001).

RESULTS: Time-kill assays revealed that HICA and Calcicur® killed all planktonic organisms within 24 h, whilst alpha-mangostin killed the organisms within 72 h. However, Odontopaste® was a slow-killing agent: 10 cells of planktonic organisms survived after exposure to the agent for 7 days. The ex vivo tooth model demonstrated that HICA and alpha-mangostin significantly inhibited the cell growth in all sampling depths (p < .0001). All species-specific data revealed the effectiveness of each medicament on the biofilm composition.

CONCLUSIONS: D,L-2-hydroxyisocaproic acid and alpha-mangostin had antimicrobial activity against multispecies bacterial-fungal biofilms.},
}

@article {pmid34469193,
year = {2021},
author = {Hashidoko, Y and Kim, D},
title = {Bidirectional cell-cell communication via indole and cyclo(Pro-Tyr) modulates interspecies biofilm formation.},
journal = {Applied and environmental microbiology},
volume = {},
number = {},
pages = {AEM0127721},
doi = {10.1128/AEM.01277-21},
pmid = {34469193},
issn = {1098-5336},
abstract = {The extracellular signaling molecule indole plays a pivotal role in biofilm formation by the enteric γ-Proteobacterium Escherichia coli; this process is particularly correlated with extracellular indole concentration. Using indole-biodegrading β-Proteobacterium Burkholderia unamae, we examined the mechanism by which these two bacteria modulate biofilm formation in an indole-dependent manner. We quantified the spatial organization of cocultured microbial communities at the micron-scale through computational image analysis, ultimately identifying how bidirectional cell-to-cell communication modulated the physical relationships between them. Further analysis allowed us to determine the mechanism by which the B. unamae-derived signaling diketopiperazine, cyclo(Pro-Tyr), considerably upregulated indole biosynthesis and enhanced E. coli biofilm formation. We also determined that the presence of unmetabolized indole enhanced production of cyclo(Pro-Tyr). Thus, bidirectional cell-to-cell communication that occurred via interspecies signaling molecules modulated formation of a mixed-species biofilm between indole-producing and indole-consuming species. Importance Indole is a relatively stable N-heterocyclic aromatic compound that is widely found in nature. To date, the correlations between indole-related bidirectional cell-to-cell communications and interspecies communal organization remain poorly understood. In this study, we used an experimental model, which consisted of indole-producing and indole-degrading bacteria, to evaluate how bidirectional cell-to-cell communication modulated interspecies biofilm formation via intrinsic and environmental cues. We identified a unique spatial patterning of indole-producing and indole-degrading bacteria within mixed-species biofilms. This spatial patterning was an active process mediated by bidirectional physico-chemical interactions. Our findings represent an important step in gaining a more thorough understanding of the process of polymicrobial biofilm formation and advance the possibility of using indole degrading bacteria to address biofilm-related health and industry issues.},
}