@article {pmid32087316,
year = {2020},
author = {Shehabeldine, A and Ashour, RM and Okba, MM and Saber, FR},
title = {Callistemon citrinus bioactive metabolites as new inhibitors of methicillin-resistant Staphylococcus aureus biofilm formation.},
journal = {Journal of ethnopharmacology},
volume = {},
number = {},
pages = {112669},
doi = {10.1016/j.jep.2020.112669},
pmid = {32087316},
issn = {1872-7573},
abstract = {The development of new anti-virulence drugs from natural origin has recently received significant attention. Callistemon citrinus Skeels is an important plant of great medicinal value. Its antimicrobial activity is well documented. Although several bioactive compounds were isolated from this plant, the actual bioactive compounds responsible for its antimicrobial activity still unrevealed.

AIM OF THE STUDY: To evaluate the effect of C. citrinus crude extract and isolates on methicillin-resistant and sensitive Staphylococcus aureus.

MATERIALS AND METHODS: C. citrinus leaves methylene chloride-methanol extract (MME) was prepared by Soxhlet apparatus. Biologically guided fractionation of MME was accomplished using several normal and reversed phase silica gel columns. The potency of MME and isolates against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) was evaluated. The mechanism of resistance was studied using three virulence factors; antibiofilm activity, inhibition of staphyloxanthin biosynthesis and effect on acid tolerance.

RESULTS: Pulverulentone A (C1), 8- desmethyl eucalyptin (C2) and eucalyptin (C3) were isolated from the most bioactive fraction of MME. Pulverulentone (C1) showed the most potent anti-biofilm activity up to 71% and 62.3% against MRSA and MSSA, respectively. It also exhibited the highest inhibition of staphyloxanthin biosynthesis of MRSA and MSSA by 55.6% and 54.5%, respectively.

CONCLUSIONS: C. citrinus phenolics and acylphloroglucinols may serve as potential source of plant-based antibacterials.},
}

@article {pmid32086747,
year = {2020},
author = {Ahmed, T and Pattnaik, S and Khan, MB and Ampasala, DR and Busi, S and Sarma, VV},
title = {Inhibition of quorum sensing-associated virulence factors and biofilm formation in Pseudomonas aeruginosa PAO1 by Mycoleptodiscus indicus PUTY1.},
journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]},
volume = {},
number = {},
pages = {},
doi = {10.1007/s42770-020-00235-y},
pmid = {32086747},
issn = {1678-4405},
abstract = {Pseudomonas aeruginosa is the second most emerging multidrug-resistant, opportunistic pathogen after Acinetobacter baumannii that poses a threat in nursing homes, hospitals, and patients who need devices such as ventilators and blood catheters. Its ability to form quorum sensing-regulated virulence factors and biofilm makes it more resistant to top most therapeutic agents such as carbapenems and next-generation antibiotics. In the current study, we studied the quorum quenching potential of secondary metabolites of Mycoleptodiscus indicus PUTY1 strain. In vitro observation showed a mitigation in virulence factors such as rhamnolipids, protease, elastase pyocyanin, exopolysaccharides, and hydrogen cyanide gas. Furthermore, a significant reduction in the motility such as swimming, swarming, twitching, and inhibition in biofilm formation by Pseudomonas aeruginosa PAO1 was observed. Results of in vitro studies were further confirmed by in silico studies through docking and molecular dynamic simulation of GC-MS-detected compounds of Mycoleptodiscus indicus employing LasR and RhlR proteins. Both in vitro and in silico observations indicate a new alternative approach for combating virulence of Pseudomonas aeruginosa by targeting its protein receptors LasR and RhlR. Graphical abstract.},
}

@article {pmid32086003,
year = {2019},
author = {Zhai, S and Ji, M and Zhao, Y and Su, X},
title = {Shift of bacterial community and denitrification functional genes in biofilm electrode reactor in response to high salinity.},
journal = {Environmental research},
volume = {184},
number = {},
pages = {109007},
doi = {10.1016/j.envres.2019.109007},
pmid = {32086003},
issn = {1096-0953},
abstract = {High salinity suppresses denitrification by inhibiting microorganism activities. The shift of microbial community and denitrification functional genes under salinity gradient was systematically investigated in a biofilm electrode reactor (BER) and biofilm reactor (BR) systems. Denitrification efficiency of both BER and BR was not significantly inhibited during the period of low salinity (0-2.0%). As the salinity increased to 2.5%, BER could overcome the impact of high salinity and maintained a relatively stable denitrification performance, and the effluent NO3--N was lower than 1.5 mg/L. High salinity (>2.5%) impoverished microbial diversity and altered the microbial community in both BER and BR. However, two genera Methylophaga and Methyloexplanations were enriched in BER due to electrochemical stimulation, which can tolerate high salinity (>3.0%). The relative abundance of Methylophaga in BER was almost 10 times as much as in BR. Paracoccus is a hydrogen autotrophic denitrifier, which was obviously inhibited with 1.0% NaCl. The hetertrophic denitrifiers were primarily responsible for the nitrate removal in the BER compared to the autotrophic denitrifiers. The abundance and proportion of denitrifying functional genes confirmed that main denitrifiers shift to salt-tolerant species (nirK-type denitrifiers) to reduce the toxic effects. The napA (2.2 × 108 to 6.5 × 108 copies/g biofilm) and nosZ (2.2 × 107 to 4.4 × 107 copies/g biofilm) genes were more abundant in BER compared to BR's, which was attributed to the enrichment of Methylophaga alcalica and Methyloversatilis universalis FAM5 in the BER. The results proved that BER had greater denitrification potential under high salinity (>2.0%) stress at the molecular level.},
}

@article {pmid32085686,
year = {2020},
author = {Cui, F and Kim, M and Lee, W and Park, C and Kim, M},
title = {Pseudo-analytical solutions for multi-species biofilm model of aerobic granular sludge.},
journal = {Environmental technology},
volume = {},
number = {},
pages = {1-29},
doi = {10.1080/09593330.2020.1733673},
pmid = {32085686},
issn = {1479-487X},
abstract = {This paper demonstrates modelling of the aerobic granular sludge (AGS) process with the pseudo-analytical solutions (PAS) of a biofilm model. A MATLAB programmed graphical user interface platform was developed to facilitate the model calculation and access. Model calibration and validation were carried out through using experimental data collected from a granular sludge sequencing batch reactor operation. The experimental and modelling results identified the distribution of heterotrophs and nitrifiers on the AGS and its contribution to the performance of wastewater treatment. The model could describe multi-species biofilms according to the distinguishing features among the three levels of PAS models. The models demonstrated increasing degrees of interaction (no interaction, competition for nitrogen and layering and protection) between heterotrophs and nitrifiers. Modelling the AGS process using PAS increases the accessibility of the simulation of multiple species in both biofilm and suspended biomass.},
}

@article {pmid32084407,
year = {2020},
author = {Lories, B and Roberfroid, S and Dieltjens, L and De Coster, D and Foster, KR and Steenackers, HP},
title = {Biofilm Bacteria Use Stress Responses to Detect and Respond to Competitors.},
journal = {Current biology : CB},
volume = {},
number = {},
pages = {},
doi = {10.1016/j.cub.2020.01.065},
pmid = {32084407},
issn = {1879-0445},
abstract = {Bacteria use complex regulatory networks to cope with stress, but the function of these networks in natural habitats is poorly understood. The competition sensing hypothesis states that bacterial stress response systems can serve to detect ecological competition, but studying regulatory responses in diverse communities is challenging. Here, we solve this problem by using differential fluorescence induction to screen the Salmonella Typhimurium genome for loci that respond, at the single-cell level, to life in biofilms with competing strains of S. Typhimurium and Escherichia coli. This screening reveals the presence of competing strains drives up the expression of genes associated with biofilm matrix production (CsgD pathway), epithelial invasion (SPI1 invasion system), and, finally, chemical efflux and antibiotic tolerance (TolC efflux pump and AadA aminoglycoside 3-adenyltransferase). We validate that these regulatory changes result in the predicted phenotypic changes in biofilm, mammalian cell invasion, and antibiotic tolerance. We further show that these responses arise via activation of major stress responses, providing direct support for the competition sensing hypothesis. Moreover, inactivation of the type VI secretion system (T6SS) of a competitor annuls the responses to competition, indicating that T6SS-derived cell damage activates these stress response systems. Our work shows that bacteria use stress responses to detect and respond to competition in a manner important for major phenotypes, including biofilm formation, virulence, and antibiotic tolerance.},
}

@article {pmid32084213,
year = {2020},
author = {Jalal, N and Lee, SF},
title = {The MsrAB reducing pathway of Streptococcus gordonii is needed for oxidative stress tolerance, biofilm formation, and oral colonization in mice.},
journal = {PloS one},
volume = {15},
number = {2},
pages = {e0229375},
doi = {10.1371/journal.pone.0229375},
pmid = {32084213},
issn = {1932-6203},
abstract = {The ability of Streptococcus gordonii to cope with oxidative stress is important for survival and persistence in dental plaque. In this study, we used mutational, phenotypic, and biochemical approaches to characterize the role of a methionine sulfoxide reductase (MsrAB) and proteins encoded by genes in the msrAB operon and an adjacent operon in oxidative stress tolerance in S. gordonii. The results showed that MsrAB and four other proteins encoded in the operons are needed for protection from H2O2 and methionine sulfoxide. These five proteins formed a reducing pathway that was needed for oxidative stress tolerance, biofilm formation, and oral colonization in mice. In the pathway, MsrAB was the enzyme that repaired oxidatively damaged proteins, and the two thioredoxin-like lipoproteins (SdbB and Sgo_1177) and two CcdA proteins were proteins that maintained the catalytic cycle of MsrAB. Consistent with the role in oxidative stress tolerance, the production of MsrAB, SdbB, and Sgo_11777 was induced in aerobic growth and planktonic cells.},
}

@article {pmid32084198,
year = {2020},
author = {Kirchhoff, L and Arweiler-Harbeck, D and Arnolds, J and Hussain, T and Hansen, S and Bertram, R and Buer, J and Lang, S and Steinmann, J and Höing, B},
title = {Imaging studies of bacterial biofilms on cochlear implants-Bioactive glass (BAG) inhibits mature biofilm.},
journal = {PloS one},
volume = {15},
number = {2},
pages = {e0229198},
doi = {10.1371/journal.pone.0229198},
pmid = {32084198},
issn = {1932-6203},
abstract = {The capability of Pseudomonas aeruginosa and Staphylococcus aureus to form biofilm on varying CI component materials differs in the presence and absence of bioactive glass (BAG). The application of BAG induces significant changes in biofilm morphology which can be visualized via scanning electron microscopy (SEM). Bacterial biofilm formation on medical devices, such as cochlear implants (CI), can lead to chronic infections. Interestingly, BAG of type S53P4 seems to be a promising tool for use in the reduction of biofilm development. Primarily, four bacterial species known to cause implant-related infections, P.aeruginosa (ATCC9027), S. aureus (ATCC6538), Staphylococcus epidermidis (ATCC12228) and Streptococcus pyogenes (ATCC19615) were analyzed regarding their capacity to form biofilm on CI components manufactured from three kinds of material: silicone, platinum and titanium. Subsequently, P. aeruginosa and S. aureus biofilms were visualized using scanning electron microscopy, comparing BAG-treated biofilm with non-treated biofilm. The four bacterial species presented biofilm-forming capabilities in a species and surface dependent manner. Metal CI components allowed for the greatest proliferation of biofilm. S. aureus and P. aeruginosa showed the highest rate of biofilm formation on polystyrene surfaces. For both species, SEM revealed altered biofilm morphology after treatment of S53P4 BAG. This study indicates that bacterial biofilm formation and structure on CI components is dependent on the surface composition, altering between metal and silicone surfaces. After application of BAG, changes in biofilm morphology on CI components were observed. These data highlight the impact of BAG on bacterial biofilm morphology.},
}

@article {pmid32083796,
year = {2020},
author = {Budell, WC and Germain, GA and Janisch, N and McKie-Krisberg, Z and Jayaprakash, AD and Resnick, AE and Quadri, LEN},
title = {Transposon mutagenesis in Mycobacterium kansasii links a small RNA gene to colony morphology and biofilm formation and identifies 9,885 intragenic insertions that do not compromise colony outgrowth.},
journal = {MicrobiologyOpen},
volume = {},
number = {},
pages = {e988},
doi = {10.1002/mbo3.988},
pmid = {32083796},
issn = {2045-8827},
support = {R03 AI 128314/NH/NIH HHS/United States ; R03 AI 135755/NH/NIH HHS/United States ; },
abstract = {Mycobacterium kansasii (Mk) is a resilient opportunistic human pathogen that causes tuberculosis-like chronic pulmonary disease and mortality stemming from comorbidities and treatment failure. The standard treatment of Mk infections requires costly, long-term, multidrug courses with adverse side effects. The emergence of drug-resistant isolates further complicates the already challenging drug therapy regimens and threatens to compromise the future control of Mk infections. Despite the increasingly recognized global burden of Mk infections, the biology of this opportunistic pathogen remains essentially unexplored. In particular, studies reporting gene function or generation of defined mutants are scarce. Moreover, no transposon (Tn) mutagenesis tool has been validated for use in Mk, a situation limiting the repertoire of genetic approaches available to accelerate the dissection of gene function and the generation of gene knockout mutants in this poorly characterized pathogen. In this study, we validated the functionality of a powerful Tn mutagenesis tool in Mk and used this tool in conjunction with a forward genetic screen to establish a previously unrecognized role of a conserved mycobacterial small RNA gene of unknown function in colony morphology features and biofilm formation. We also combined Tn mutagenesis with next-generation sequencing to identify 12,071 Tn insertions that do not compromise viability in vitro. Finally, we demonstrated the susceptibility of the Galleria mellonella larva to Mk, setting the stage for further exploration of this simple and economical infection model system to the study of this pathogen.},
}

@article {pmid32083136,
year = {2020},
author = {Li, X and Kim, J and Wu, J and Ahamed, AI and Wang, Y and Martins-Green, M},
title = {N-Acetyl-cysteine and Mechanisms Involved in Resolution of Chronic Wound Biofilm.},
journal = {Journal of diabetes research},
volume = {2020},
number = {},
pages = {9589507},
doi = {10.1155/2020/9589507},
pmid = {32083136},
issn = {2314-6753},
abstract = {Chronic wounds are a major global health problem with the presence of biofilm significantly contributing to wound chronicity. Current treatments are ineffective in resolving biofilm and simultaneously killing the bacteria; therefore, effective biofilm-resolving drugs are needed. We have previously shown that, together with α-tocopherol, N-acetyl-cysteine (NAC) significantly improves the healing of biofilm-containing chronic wounds, in a diabetic mouse model we developed, by causing disappearance of the bacteria and breakdown of the extracellular polymeric substance (EPS). We hypothesize that NAC creates a microenvironment that affects bacterial survival and EPS integrity. To test this hypothesis, we developed an in vitro biofilm system using microbiome taken directly from diabetic mouse chronic wounds. For these studies, we chose mice in which chronic wound microbiome was rich in Pseudomonas aeruginosa (97%). We show that NAC at concentrations with pH < pKa causes bacterial cell death and breakdown of EPS. When used before biofilm is formed, NAC leads to bacterial cell death whereas treatment after the biofilm is established NAC causes biofilm dismantling accompanied by bacterial cell death. Mechanistically, we show that NAC can penetrate the bacterial membrane, increase oxidative stress, and halt protein synthesis. We also show that low pH is important for the actions of NAC and that bacterial death occurs independently of the presence of biofilm. In addition, we show that both the acetyl and carboxylic groups play key roles in NAC functions. The results presented here provide insight into the mechanisms by which NAC dismantles biofilm and how it could be used to treat chronic wounds after debridement (NAC applied at the start of culture) or without debridement (NAC applied when biofilm is already formed). This approach can be taken to develop biofilm from microbiome taken directly from human chronic wounds to test molecules that could be effective for the treatment of specific biofilm compositions.},
}

@article {pmid32082279,
year = {2020},
author = {Martín-Rodríguez, AJ and Rhen, M and Melican, K and Richter-Dahlfors, A},
title = {Nitrate Metabolism Modulates Biosynthesis of Biofilm Components in Uropathogenic Escherichia coli and Acts as a Fitness Factor During Experimental Urinary Tract Infection.},
journal = {Frontiers in microbiology},
volume = {11},
number = {},
pages = {26},
doi = {10.3389/fmicb.2020.00026},
pmid = {32082279},
issn = {1664-302X},
abstract = {To successfully colonize a variety of environments, bacteria can coordinate complex collective behaviors such as biofilm formation. To thrive in oxygen limited niches, bacteria's versatile physiology enables the utilization of alternative electron acceptors. Nitrate, the second most favorable electron acceptor after oxygen, plays a prominent role in the physiology of uropathogenic Escherichia coli (UPEC) and is abundantly found in urine. Here we analyzed the role of extracellular nitrate in the pathogenesis of the UPEC strain CFT073 with an initial focus on biofilm formation. Colony morphotyping in combination with extensive mutational, transcriptional, and protein expression analyses of CFT073 wild-type and mutants deficient in one or several nitrate reductases revealed an association between nitrate reduction and the biosynthesis of biofilm extracellular matrix components. We identified a role for the nitrate response regulator NarL in modulating expression of the biofilm master regulator CsgD. To analyze the role of nitrate reduction during infection in vivo, we tested wild-type CFT073 and a nitrate reductase null mutant in an ascending urinary tract infection (UTI) model. Individually, each strain colonized extensively, suggesting that nitrate reduction is expendable during UTI. However, during competitive co-infection, the strain incapable of nitrate reduction was strongly outcompeted. This suggests that nitrate reduction can be considered a non-essential but advantageous fitness factor for UPEC pathogenesis. This implies that UPEC rapidly adapts their metabolic needs to the microenvironment of infected tissue. Collectively, this work demonstrates a unique association between nitrate respiration, biofilm formation, and UPEC pathogenicity, highlighting how the use of alternative electron acceptors enables bacterial pathogens to adapt to challenging infectious microenvironments.},
}

@article {pmid32081907,
year = {2020},
author = {Tursi, SA and Puligedda, RD and Szabo, P and Nicastro, LK and Miller, AL and Qiu, C and Gallucci, S and Relkin, NR and Buttaro, BA and Dessain, SK and Tükel, Ç},
title = {Salmonella Typhimurium biofilm disruption by a human antibody that binds a pan-amyloid epitope on curli.},
journal = {Nature communications},
volume = {11},
number = {1},
pages = {1007},
doi = {10.1038/s41467-020-14685-3},
pmid = {32081907},
issn = {2041-1723},
abstract = {Bacterial biofilms, especially those associated with implanted medical devices, are difficult to eradicate. Curli amyloid fibers are important components of the biofilms formed by the Enterobacteriaceae family. Here, we show that a human monoclonal antibody with pan-amyloid-binding activity (mAb 3H3) can disrupt biofilms formed by Salmonella enterica serovar Typhimurium in vitro and in vivo. The antibody disrupts the biofilm structure, enhancing biofilm eradication by antibiotics and immune cells. In mice, 3H3 injections allow antibiotic-mediated clearance of catheter-associated S. Typhimurium biofilms. Thus, monoclonal antibodies that bind a pan-amyloid epitope have potential to prevent or eradicate bacterial biofilms.},
}

@article {pmid32077490,
year = {2020},
author = {Islam, M and Durie, I and Ramadan, R and Purchase, D and Marvasi, M},
title = {Exploitation of nitric oxide donors to control bacterial adhesion on ready-to-eat vegetables and dispersal of pathogenic biofilm from polypropylene.},
journal = {Journal of the science of food and agriculture},
volume = {},
number = {},
pages = {},
doi = {10.1002/jsfa.10340},
pmid = {32077490},
issn = {1097-0010},
abstract = {BACKGROUND: Nitric oxide (NO) donors have been used to control biofilm formation. NO can be delivered in situ using organic carriers and act as a signaling molecule. Cells exposed to NO shift from biofilm to the planktonic state and are better exposed to the action of disinfectants. In this study, we investigate the capability of the NO donors molsidomine, MAHAMA NONOate, NO-aspirin and diethylamine NONOate to act as anti-adhesion agents on ready-to-eat vegetables, as well as dispersants to a number of pathogenic biofilms on plastic.

RESULTS: Our results showed that 10pM molsidomine reduced the attachment of Salmonella enterica sv Typhimurium 14 028 to pea shoots and coriander leaves of about 0.5 Log(CFU/leaf) when compared with untreated control. The association of 10 pM molsidomine with 0.006% H2 O2 showed a synergistic effect, obtaining a significant reduction in cell collection on the surface of the vegetable of about 1 Log(CFU/leaf). Similar results were obtained for MAHMA NONOate. We also showed that the association of diethylamine NONOate at 10 mM and 10pM with the quaternary ammonium compound diquat bromide improves the effectiveness of biofilm dispersal by 50% when compared with the donor alone.

CONCLUSIONS: Our findings reveal the dual role of NO compounds in biofilm control. Molsidomine, MAHMA NONOate and diethylamine NONOate are good candidates in either preventing biofilm formation or dispersing biofilm, especially when used in conjunction with disinfectants. NO compounds have the potential to be developed into tool-kit for pro-active practices for GAPs, HACCP and Cleaning-in-place (CIP) protocols in industrial settings where washing is routinely applied. This article is protected by copyright. All rights reserved.},
}

@article {pmid32077074,
year = {2020},
author = {Reginatto, P and Bergamo, VZ and Berlitz, SJ and Guerreiro, ICK and de Andrade, SF and Fuentefria, AM},
title = {Rational selection of antifungal drugs to propose a new formulation strategy to control Candida biofilm formation on venous catheters.},
journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]},
volume = {},
number = {},
pages = {},
doi = {10.1007/s42770-020-00242-z},
pmid = {32077074},
issn = {1678-4405},
abstract = {INTRODUCTION: Infections associated with medical devices are often related to colonization by Candida spp. biofilm; in this way, numerous strategies have been developed and studied, mainly in order to prevent this type of fungal growth.

AIM: Considering the above, the main objective of the present study is to make a rational choice of the best antifungal therapy for the in vitro treatment of the biofilm on venous catheters, proposing an innovative formulation of a film-forming system to coat the surface in order to prevent the formation of biofilms.

METHODOLOGY: Anidulafungin, fluconazole, voriconazole, ketoconazole, amphotericin B, and the association of anidulafungin and amphotericin B were tested against biofilms of C. albicans, C. tropicalis, and C. parapsilosis strains in microtiter plates and in a polyurethane catheter. Besides, anidulafungin, amphotericin B, and the combination of both were incorporated in a film-forming system and were evaluated against biofilm.

RESULTS: The superior activity of anidulafungin was demonstrated in relation to the other antifungal agents. Although amphotericin B showed good activity, high concentrations were required. The combination showed a synergistic action, in solution and in the formulation, showing excellent results, with activity above 90%.

CONCLUSION: Due to the superiority of anidulafungin and the synergistic activity of the combination, these alternatives were the most promising options for use in a formulation proposal as a new strategy to combat the Candida spp. biofilm. These formulations demonstrated high in vitro performance in the prevention of biofilms, indicating that they are candidates with great potential for in vivo tests.},
}

@article {pmid32076613,
year = {2020},
author = {Feldman, M and Sionov, R and Smoum, R and Mechoulam, R and Ginsburg, I and Steinberg, D},
title = {Comparative Evaluation of Combinatory Interaction between Endocannabinoid System Compounds and Poly-L-lysine against Streptococcus mutans Growth and Biofilm Formation.},
journal = {BioMed research international},
volume = {2020},
number = {},
pages = {7258380},
doi = {10.1155/2020/7258380},
pmid = {32076613},
issn = {2314-6141},
abstract = {Endocannabinoid/endocannabinoid-like (EC/EC-like) are natural endogenous compounds which have been found to affect MRSA pathogenicity. Our previous studies showed that EC/EC-like was able to impair staphylococcal biofilm formation and maintenance as well as to alter biofilm-associated virulence factors. In the present study, we investigated the combinatory effect of the selected EC/EC-like with a natural antimicrobial agent, poly-L-lysine, on cariogenic bacteria Streptococcus mutans growth and biofilm formation. Among four tested EC/EC-like, only two, anandamide (AEA) and oleoylethanolamide (OEA), exhibited synergistic combinatory effect with poly-L-lysine against S. mutans. We attribute this distinct effect to differences in the fatty acid chain structure of the selected EC/EC-like compounds. Moreover, AEA exerted a specific antibiofilm mode of action against S. mutans by effecting total inhibition of biofilm formation while still allowing bacteria viability. Finally, we postulate that the presence of EC/EC-like and poly-L-lysine could enhance the permeability and efficacy of each other via hydrophobic and electrostatic interactions with the S. mutans membrane. In conclusion, we assume that a combination of endogenous natural compounds such as EC/EC-like and poly-L-lysine may benefit oral hygiene by preventing dental plaque.},
}

@article {pmid32074168,
year = {2020},
author = {Shafiei, SNS and Ahmad, K and Ikhsan, NFM and Ismail, SI and Sijam, K},
title = {Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract.},
journal = {Brazilian journal of biology = Revista brasleira de biologia},
volume = {},
number = {},
pages = {},
doi = {10.1590/1519-6984.206124},
pmid = {32074168},
issn = {1678-4375},
abstract = {Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.},
}

@article {pmid32074086,
year = {2019},
author = {Goudarzi, M and Mohammadi, A and Amirpour, A and Fazeli, M and Nasiri, MJ and Hashemi, A and Goudarzi, H},
title = {Genetic diversity and biofilm formation analysis of Staphylococcus aureus causing urinary tract infections in Tehran, Iran.},
journal = {Journal of infection in developing countries},
volume = {13},
number = {9},
pages = {777-785},
doi = {10.3855/jidc.11329},
pmid = {32074086},
issn = {1972-2680},
abstract = {INTRODUCTION: Over the past decades, prevalence of biofilm-forming Staphylococcus aureus strains has significantly increased in urinary tract infections. The aim of this study was to investigate prevalence of biofilm forming and adhesion encoding genes and to analyze distribution of different agr and spa types in S. aureus isolates.

METHODOLOGY: In the present study, 75 S. aureus isolates obtained from patients with urinary tract infections were examined for susceptibility to antimicrobial agents. Adhesion, biofilm, and spa encoding genes were detected by PCR screening; agr types were determined using multiplex PCR.

RESULTS: Among the 75 isolates, 72% were biofilm producers and 28% were non-biofilm producers. Notably, the ability to produce biofilm was higher among MRSA strains ompared to MSSA strains. The most prevalent biofilm forming gene was icaD (77.3%), followed by icaA (76%), icaB (57.3%) and icaC (50.7%). Adhesion genes clfA, clfB, fnbB, can, fnbA, ebp and bap were detected in 94.7%, 92%, 68%, 64%, 64%, 60% and 5.3% of the isolates, respectively. The spa types t426 and t7789 were found among the non-MDR isolates. It was found that t790, t084, t7789 and t325 spa types were biofilm producers, while t426 and t1339 spa types were non-biofilm producers.

CONCLUSION: Biofilm encoding genes icaD and spa type t790 and agr type III were the most prevalent factors among MDR biofilm producer isolates. The study emphasized that identification of genes and characterization of molecular types involved in biofilm formation should be considered.},
}

@article {pmid32072274,
year = {2020},
author = {Ren, X and Wang, L and Chen, W},
title = {Oxytropis glabra DC. Inhibits Biofilm Formation of Staphylococcus epidermidis by Down-Regulating ica Operon Expression.},
journal = {Current microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1007/s00284-019-01847-w},
pmid = {32072274},
issn = {1432-0991},
support = {Grant 31260026//the National Science Foundation of China/ ; Grant NCET-11-1071//Excellent Young Talents Fund Program of Higher Education Institutions of Anhui Province (CN)/ ; Grant 2009JC07//Society of the Friendly Sons of St. Patrick for the Relief of Emigrants from Ireland (US)/ ; },
abstract = {Staphylococcus epidermidis is one of the main causes of medical device-related infections and bovine mastitis owing to its biofilm-forming abilities. Oxytropis glabra DC. is one of the most widespread Fabaceae species and used as a Chinese herbal formulation in Western China. Our research investigated the effects of O. glabra on the biofilm formation of S. epidermidis and the possible inhibiting mechanism. The biofilm-forming reference strain, S. epidermidis SE-1 (ATCC 35,984), was employed as a model and semi-quantitative biofilm assay was performed to evaluate the antibiofilm activity of O. glabra. The exopolysaccharides (EPS) production and expression of ica operon were studied to explore the possible antibiofilm mechanism using thin-layer chromatography and quantitative real-time PCR assay, respectively. The results obtained indicated that O. glabra decoction at 7.5 mg mL-1 significantly inhibited biofilm formation by about 95% without affecting cell growth of S. epidermidis. Two hydrolysis productions of EPS were significantly decreased by 64% and 54% with the addition of 7.5 mg mL-1O. glabra and the expression of icaR was significantly up-regulated 2.2-times, whereas icaB was significantly down-regulated more than 50% by 7.5 mg mL-1O. glabra. These findings suggest a potential application for O. glabra as a promising candidate for the exploration of new drugs against S. epidermidis biofilm-associated infections.},
}

@article {pmid32071069,
year = {2020},
author = {Sun, Y and Li, Y and Luo, Q and Huang, J and Chen, J and Zhang, R and Wang, X},
title = {LuxS/AI-2 Quorum Sensing System in Edwardsiella piscicida promotes its biofilm formation and pathogenicity.},
journal = {Infection and immunity},
volume = {},
number = {},
pages = {},
doi = {10.1128/IAI.00907-19},
pmid = {32071069},
issn = {1098-5522},
abstract = {LuxS/AI-2 is an important quorum sensing system, which affects the growth, biofilm formation, virulence and metabolism of bacteria. LuxS is encoded by luxS gene, but how this gene is associated with a diverse array of physiological activities in Edwardsiella piscicida (E. piscicida) are not available. Here we constructed the luxS gene mutant strain named ΔluxS to identify LuxS/AI-2 how to affect the pathogenicity. The result showed that LuxS was not found in the luxS gene mutant strain, and this gene deletion decreases E. piscicida growth compare to the wild-type strain. Meanwhile, the wild-type strain significantly increased penetration and motility in mucin compared to the ΔluxS. The LD50 of E. piscicida ΔluxS strain for zebrafish was significantly higher than the wild-type strain, which suggested the luxS gene deletion could attenuate its virulence. The AI-2 activities of EIB202 were 56-fold higher than the ones in the ΔluxS strain, suggested the luxS gene promotes AI-2 production. Transcriptome results demonstrated that 46 significantly differential genes were achieved between cells infected with ΔluxS and the wild-type strain, which included 34 up-regulated genes and 12 down-regulated genes. Among those genes, the largest parts were closely related to cell immunity and signaling system. Besides, the biofilm formation ability of EIB202 was significantly higher than that of ΔluxS, meanwhile the supernatant of EIB202 increased the biofilm formation ability of ΔluxS, which suggested that the luxS gene and its product LuxS enhanced biofilm formation in E. piscicida All results indicate that the LuxS/AI-2 quorum-sensing system in E. piscicida promotes its pathogenicity through increasing a diverse array of physiological activities.},
}

@article {pmid32070890,
year = {2020},
author = {Shabbir, S and Faheem, M and Ali, N and Kerr, PG and Wang, LF and Kuppusamy, S and Li, Y},
title = {Periphytic biofilm: An innovative approach for biodegradation of microplastics.},
journal = {The Science of the total environment},
volume = {717},
number = {},
pages = {137064},
doi = {10.1016/j.scitotenv.2020.137064},
pmid = {32070890},
issn = {1879-1026},
abstract = {Microplastics (MPs) have been gaining the attention of environmental researchers since the 1960s anecdotal reports of plastic entanglement and ingestion by marine creatures. Due to their increasing accretion in aquatic environments, as well as resistance towards degradation, marine litter research has focused on microplastics more recently. In the present study, a relatively new method of biodegradation was implemented for the biodegradation of three structurally different MPs i.e. polypropylene (PP), polyethylene (PE) and polyethylene terephthalate (PET). Periphytic biofilm was used for this purpose in various backgrounds of carbon sources (glucose, peptone, and glucose and peptone). Biodegradation of MPs was estimated in terms of weight loss. It was observed that the addition of glucose enhanced the biodegradation of MPs by periphyton biofilm for all MPs (from 9.52%-18.02%, 5.95%-14.02% and 13.24-19.72% for PP, PE and PET respectively) after 60 days compared to natural biofilm alone. To the contrary, peptone, and glucose and peptone together, were inhibitory. Biodegradation was further confirmed by morphological changes observed using SEM, FTIR spectra and GPC lent further support to the results whereby new peaks appeared along with reduction in old peaks and decrease in peak intensities. MiSeq sequencing shows that Deinococcus-thermus > Proteobacteria > Cyanobacteria are the dominant phyla in natural biofilms, and their relative abundances increase after the addition of glucose. However, the abundances shifted to Deinococcus-thermus > Cyanobacteria > Firmicutes > Bacteroidetes, when the biofilms were treated with either peptone alone, or with glucose and peptone together. Therefore, the change in biodegradation capability might also be due to the change in the microbial community structures after addition of the C-sources. These experiments provide an innovative approach towards effective biodegradation of MPs using a relatively new environment-friendly method.},
}

@article {pmid32070273,
year = {2020},
author = {Chen, T and Dong, G and Zhang, S and Zhang, X and Zhao, Y and Cao, J and Zhou, T and Wu, Q},
title = {Effects of iron on the growth, biofilm formation and virulence of Klebsiella pneumoniae causing liver abscess.},
journal = {BMC microbiology},
volume = {20},
number = {1},
pages = {36},
doi = {10.1186/s12866-020-01727-5},
pmid = {32070273},
issn = {1471-2180},
support = {No.81971986//National Natural Science Foundation of China/ ; No.2019KY098//Health Department of Zhejiang Province of the People's Republic of China/ ; },
abstract = {BACKGROUND: Klebsiella pneumoniae is considered the most clinically relevant species of Enterobacteriaceae, known to cause severe infections including liver abscesses. To the best of our knowledge, a large proportion of iron in the human body is accumulated and stored in the liver. We hypothesize that increased iron availability is an important factor driving liver abscess formation and we therefore aim to understand the effects of iron on K. pneumoniae causing liver abscesses.

RESULTS: All tested K. pneumoniae clinical isolates, including those isolated from liver abscesses and other abdominal invasive infection sites, grew optimally when cultured in LB broth supplemented with 50 μM iron and exhibited the strongest biofilm formation ability under those conditions. Decreased growth and biofilm formation ability were observed in all tested strains when cultured with an iron chelator (P < 0.05). The infection model of G. mellonella larvae indicated the virulence of liver abscess-causing K. pneumoniae (2/3) cultured in LB broth with additional iron was significantly higher than those under iron-restricted conditions (P < 0.05). The relative expression levels of the four siderophore genes (iucB, iroB, irp1, entB) in K. pneumoniae strains isolated from liver abscesses cultured with additional iron were lower than those under iron-restricted conditions (P < 0.05).

CONCLUSIONS: It is suggested by our research that iron in the environment can promote growth, biofilm formation and enhance virulence of K. pneumoniae causing liver abscesses. A lower expression of siderophore genes correlates with increased virulence of liver abscess-causing K. pneumoniae. Further deeper evaluation of these phenomena is warranted.},
}

@article {pmid32070067,
year = {2020},
author = {Hahn, MM and Gunn, JS},
title = {Salmonella Extracellular Polymeric Substances Modulate Innate Phagocyte Activity and Enhance Tolerance of Biofilm-Associated Bacteria to Oxidative Stress.},
journal = {Microorganisms},
volume = {8},
number = {2},
pages = {},
doi = {10.3390/microorganisms8020253},
pmid = {32070067},
issn = {2076-2607},
support = {R01AI116917/NH/NIH HHS/United States ; },
abstract = {Salmonella enterica serovar Typhi causes 14.3 million acute cases of typhoid fever that are responsible for 136,000 deaths each year. Chronic infections occur in 3%-5% of those infected and S. Typhi persists primarily in the gallbladder by forming biofilms on cholesterol gallstones, but how these bacterial communities evade host immunity is not known. Salmonella biofilms produce several extracellular polymeric substances (EPSs) during chronic infection, which are hypothesized to prevent pathogen clearance either by protecting biofilm-associated bacteria from direct humoral attack or by modulating innate phagocyte interaction with biofilms. Using wild-type and EPS-deficient planktonic and biofilm Salmonella, the direct attack hypothesis was tested by challenging biofilms with human serum and antimicrobial peptides. Biofilms were found to be tolerant to these molecules, but these phenotypes were independent of the tested EPSs. By examining macrophage and neutrophil responses, new roles for biofilm-associated capsular polysaccharides and slime polysaccharides were identified. The S. Typhi Vi antigen was found to modulate innate immunity by reducing macrophage nitric oxide production and neutrophil reactive oxygen species (ROS) production. The slime polysaccharides colanic acid and cellulose were found to be immune-stimulating and represent a key difference between non-typhoidal serovars and typhoidal serovars, which do not express colanic acid. Furthermore, biofilm tolerance to the exogenously-supplied ROS intermediates hydrogen peroxide (H2O2) and hypochlorite (ClO) indicated an additional role of the capsular polysaccharides for both serovars in recalcitrance to H2O2 but not ClO, providing new understanding of the stalemate that arises during chronic infections and offering new directions for mechanistic and clinical studies.},
}

@article {pmid32069253,
year = {2019},
author = {Rodrigues, RS and Lima, NCDS and Taborda, RLM and Esquerdo, RP and Gama, AR and Nogueira, PA and Orlandi, PP and Matos, NB},
title = {Antibiotic resistance and biofilm formation in children with Enteropathogenic Escherichia coli (EPEC) in Brazilian Amazon.},
journal = {Journal of infection in developing countries},
volume = {13},
number = {8},
pages = {698-705},
doi = {10.3855/jidc.10674},
pmid = {32069253},
issn = {1972-2680},
abstract = {INTRODUCTION: Enteropathogenic Escherichia coli is an important causative agent of diarrhea in both developed and developing countries.

METHODOLOGY: We assessed the antibiotic resistance profile and the ability of 71 Enteropathogenic Escherichia coli (EPEC) isolates from children in the age group 6 years, or younger, to form biofilm. These children were hospitalized in Cosme and Damião Children Hospital in Porto Velho, Western Brazilian Amazon, between 2010 and 2012, with clinical symptoms of acute gastroenteritis.

RESULTS: The highest frequency of atypical EPEC (aEPEC) isolates reached 83.1% (59/71). Most EPEC isolates presented Localized Adherence Like (LAL) pattern in HEp-2 cells (57.7% - 41/71). Biofilm production was observed in 33.8% (24/71) of EPEC isolates, and it means statistically significant association with shf gene (p = 0.0254). The highest antimicrobial resistance rates and a large number of multiresistant isolates 67.6% (48/71), regarded cefuroxime (CXM), ampicillin (AMP), trimethoprim-sulfamethoxazole (SXT) and tetracycline (TET), respectively, mainly in typical EPEC (tEPEC). Furthermore, 96% (68/71) of EPEC isolates in the present study were resistant to at least one antibiotic, whereas only 3 isolates were sensitive to all the tested drugs.

CONCLUSION: Based on our findings, there was increased aEPEC identification. EPEC isolates showed high resistance rate; most strains showed multiresistance; thus, they work as warning about the continuous need of surveillance towards antimicrobial use. Besides, the ability of forming biofilm was evidenced by the EPEC isolates. This outcome is worrisome, since it is a natural resistance mechanism of bacteria.},
}

@article {pmid32069025,
year = {2020},
author = {Yuan, Z and Lin, C and He, Y and Tao, B and Chen, M and Zhang, J and Liu, P and Cai, K},
title = {Near Infrared Light Triggered Nitric Oxide-Enhanced Photodynamic Therapy and Low-Temperature Photothermal Therapy for Biofilm Elimination.},
journal = {ACS nano},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsnano.9b09871},
pmid = {32069025},
issn = {1936-086X},
abstract = {Photothermal treatment (PTT) involving combination therapeutic modalities were recently emerged as an efficient alternative for combating biofilm. However, PTT-related local high temperature may destroy the surrounding healthy tissues. Herein, we present an all-in-one photo-therapeutic nanoplatform consisting of L-arginine (L-Arg), indocyanine green (ICG) and mesoporous polydopamine (MPDA), namely AI-MPDA, to eliminate the already-formed biofilm. The fabrication process included surface modification of MPDA with L-Arg and further adsorption of ICG via π-π stacking. Under near infrared (NIR) exposure, AI-MPDA not only generated heat, but also produced reactive oxygen species (ROS), causing cascade catalysis of L-Arg to release nitric oxide (NO). Under near infrared (NIR) irradiation, biofilm elimination was attributed to the NO-enhanced photodynamic therapy (PDT) and low-temperature PTT (≤ 45 °C). Notably, NIR-triggered all-in-one strategy resulted in severe destruction of bacterial membranes. The photo-therapeutic AI-MPDA also displayed good cytocompatibility. NIR-irradiated AI-MPDA nanoparticles not only prevented bacterial colonization, but also realized a rapid recovery of infected wound. More importantly, the all-in-one photo-therapeutic platform displayed effective biofilm elimination with an efficiency of around 100% in a abscess formation model. Overall, this low-temperature photo-therapeutic platform provides a reliable tool for combating already-formed biofilm in clinical applications.},
}

@article {pmid32065820,
year = {2019},
author = {Nor A'shimi, MH and Alattraqchi, AG and Mohd Rani, F and A Rahman, NI and Ismail, S and Abdullah, FH and Othman, N and Cleary, DW and Clarke, SC and Yeo, CC},
title = {Biocide susceptibilities and biofilm-forming capacities of Acinetobacter baumannii clinical isolates from Malaysia.},
journal = {Journal of infection in developing countries},
volume = {13},
number = {7},
pages = {626-633},
doi = {10.3855/jidc.11455},
pmid = {32065820},
issn = {1972-2680},
abstract = {INTRODUCTION: Acinetobacter baumannii is a Gram-negative nosocomial pathogen that has the capacity to develop resistance to all classes of antimicrobial compounds. However, very little is known regarding its susceptibility to biocides (antiseptics and disinfectants) and capacity to form biofilms, particularly for Malaysian isolates.

AIM: To determine the susceptibility of A. baumannii isolates to commonly-used biocides, investigate their biofilm-forming capacities and the prevalence of biocide resistance and biofilm-associated genes.

METHODOLOGY: . The minimum inhibitory concentration (MIC) values of 100 A. baumannii hospital isolates from Terengganu, Malaysia, towards the biocides benzalkonium chloride (BZK), benzethonium chloride (BZT) and chlorhexidine digluconate (CLX), were determined by broth microdilution. The isolates were also examined for their ability to form biofilms in 96-well microplates. The prevalence of biocide resistance genes qacA, qacE and qacDE1 and the biofilm-associated genes bap and abaI were determined by polymerase chain reaction (PCR).

RESULTS: Majority of the A. baumannii isolates (43%) showed higher MIC values (> 50 µg/mL) for CLX than for BZK (5% for MIC > 50 µg/mL) and BZT (9% for MIC > 50 µg/mL). The qacDE1 gene was predominant (63%) followed by qacE (28%) whereas no isolate was found harbouring qacA. All isolates were positive for the bap and abaI genes although the biofilm-forming capacity varied among the isolates.

CONCLUSION: The Terengganu A. baumannii isolates showed higher prevalence of qacDE1 compared to qacE although no correlation was found with the biocides' MIC values. No correlation was also observed between the isolates' biofilm-forming capacity and the MIC values for the biocides.},
}

@article {pmid32064391,
year = {2020},
author = {Ripa, R and Shen, AQ and Funari, R},
title = {Detecting Escherichia coli Biofilm Development Stages on Gold and Titanium by Quartz Crystal Microbalance.},
journal = {ACS omega},
volume = {5},
number = {5},
pages = {2295-2302},
doi = {10.1021/acsomega.9b03540},
pmid = {32064391},
issn = {2470-1343},
abstract = {Bacterial biofilms are responsible for persistent infections and biofouling, raising serious concerns in both medical and industrial processes. These motivations underpin the need to develop methodologies to study the complex biological structures of biofilms and prevent their formation on medical implants, tools, and industrial apparatuses. Here, we report the detailed comparison of Escherichia coli biofilm development stages (adhesion, maturation, and dispersion) on gold and titanium surfaces by monitoring the changes in both frequency and dissipation of a quartz crystal microbalance (QCM) device, a cheap and reliable microgravimetric sensor which allows the real-time and label-free characterization of various stages of biofilm development. Although gold is the most common electrode material used for QCM sensors, the titanium electrode is also readily available for QCM sensors; thus, QCM sensors with different metal electrodes serve as a simple platform to probe how pathogens interact with different metal substrates. The QCM outcomes are further confirmed by atomic force microscopy and crystal violet staining, thus validating the effectiveness of this surface sensitive sensor for microbial biofilm research. Moreover, because QCM technology can easily modify the substrate types and coatings, QCM sensors also provide well-controlled experimental conditions to study antimicrobial surface treatments and eradication procedures, even on mature biofilms.},
}

@article {pmid32059494,
year = {2020},
author = {Fagerlund, A and Heir, E and Møretrø, T and Langsrud, S},
title = {Listeria Monocytogenes Biofilm Removal Using Different Commercial Cleaning Agents.},
journal = {Molecules (Basel, Switzerland)},
volume = {25},
number = {4},
pages = {},
doi = {10.3390/molecules25040792},
pmid = {32059494},
issn = {1420-3049},
support = {262306//Norwegian Research Funding for Agriculture and Food Industry/ ; 221663//Norwegian Research Funding for Agriculture and Food Industry/ ; },
abstract = {Effective cleaning and disinfection (C&D) is pivotal for the control of Listeria monocytogenes in food processing environments. Bacteria in biofilms are protected from biocidal action, and effective strategies for the prevention and removal of biofilms are needed. In this study, different C&D biofilm control strategies on pre-formed L. monocytogenes biofilms on a conveyor belt material were evaluated and compared to the effect of a conventional chlorinated, alkaline cleaner (agent A). Bacterial reductions up to 1.8 log were obtained in biofilms exposed to daily C&D cycles with normal user concentrations of alkaline, acidic, or enzymatic cleaning agents, followed by disinfection using peracetic acid. No significant differences in bactericidal effects between the treatments were observed. Seven-day-old biofilms were more tolerant to C&D than four-day-old biofilms. Attempts to optimize biofilm eradication protocols for four alkaline, two acidic, and one enzymatic cleaning agent, in accordance with the manufacturers' recommendations, were evaluated. Increased concentrations, the number of subsequent treatments, the exposure times, and the temperatures of the C&D agents provided between 4.0 and >5.5 log reductions in colony forming units (CFU) for seven-day-old L. monocytogenes biofilms. Enhanced protocols of conventional and enzymatic C&D protocols have the potential for improved biofilm control, although further optimizations and evaluations are needed.},
}

@article {pmid32059431,
year = {2020},
author = {Meto, A and Colombari, B and Meto, A and Boaretto, G and Pinetti, D and Marchetti, L and Benvenuti, S and Pellati, F and Blasi, E},
title = {Propolis Affects Pseudomonas aeruginosa Growth, Biofilm Formation, eDNA Release and Phenazine Production: Potential Involvement of Polyphenols.},
journal = {Microorganisms},
volume = {8},
number = {2},
pages = {},
doi = {10.3390/microorganisms8020243},
pmid = {32059431},
issn = {2076-2607},
abstract = {Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen responsible for a wide range of clinical conditions, from mild infections to life-threatening nosocomial biofilm-associated diseases, which are particularly severe in susceptible individuals. The aim of this in vitro study was to assess the effects of an Albanian propolis on several virulence-related factors of P. aeruginosa, such as growth ability, biofilm formation, extracellular DNA (eDNA) release and phenazine production. To this end, propolis was processed using three different solvents and the extracted polyphenolic compounds were identified by means of high performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) analysis. As assessed by a bioluminescence-based assay, among the three propolis extracts, the ethanol (EtOH) extract was the most effective in inhibiting both microbial growth and biofilm formation, followed by propylene glycol (PG) and polyethylene glycol 400 (PEG 400) propolis extracts. Furthermore, Pseudomonas exposure to propolis EtOH extract caused a decrease in eDNA release and phenazine production. Finally, caffeic acid phenethyl ester (CAPE) and quercetin decreased upon propolis EtOH extract exposure to bacteria. Overall, our data add new insights on the anti-microbial properties of a natural compound, such as propolis against P. aeruginosa. The potential implications of these findings will be discussed.},
}

@article {pmid32058691,
year = {2020},
author = {Konai, MM and Barman, S and Issa, R and MacNeil, S and Adhikary, U and De, K and Monk, PN and Haldar, J},
title = {Hydrophobicity Modulated Antibacterial Small Molecule Eradicates Biofilm with Potent Efficacy against Skin-infections.},
journal = {ACS infectious diseases},
volume = {},
number = {},
pages = {},
doi = {10.1021/acsinfecdis.9b00334},
pmid = {32058691},
issn = {2373-8227},
abstract = {The role of molecular arrangement of hydrophobic and hydrophilic groups for designing membrane-active molecules remains largely ambiguous. To explore this aspect, herein we report a series of membrane-active small molecules by varying the spatial distribution of hydrophobic groups. The two terminal amino groups of linear triamines such as diethylene triamine, bis(trimethylene)triamine and bis(hexamethylene)triamine were conjugated with cationic amino acids bearing variable side chain hydrophobicity (such as diaminobutyric acid, ornithine and lysine). The hydrophobicity was also modulated through conjugation of different long chain fatty acids with the central secondary amino group of the triamine. Molecules with constant backbone hydrophobicity displayed an enhanced antibacterial activity and decreased hemolytic activity upon increasing the side chain hydrophobicity of amino acids. On the other hand, increased hydrophobicity in the backbone introduced a slight hemolytic activity but a higher increment in antibacterial activity resulting in better selective antibacterial compounds. The optimized lead compound derived from structure-activity-relationship (SAR) studies was the dodecanoyl analogue of lysine series of compounds consisting of bis(hexamethylene)triamine as the backbone. This compound was active against various Gram-positive and Gram-negative bacteria at a low concentration (MIC ranged between 3.1-6.3 µg/mL) and displayed low toxicity towards mammalian cells (HC50 = 890 µg/mL and EC50 against HEK = 85 µg/mL). Additionally, it was able to kill metabolically inactive bacterial cells and eradicate preformed biofilms of MRSA. This compound showed excellent activity in a mouse model of skin-infection with reduction of ~4 log MRSA burden at 40 mg/kg dose without any sign of skin-toxicity even at 200 mg/kg. More importantly, it revealed potent efficacy in an ex-vivo model of human skin-infection (with reduction of 85% MRSA burden at 50 μg/mL), which indicates great potential of the compound as an antibacterial agent to treat skin-infections.},
}

@article {pmid32058164,
year = {2020},
author = {Dash, DM and Osborne, WJ},
title = {Rapid biodegradation and biofilm-mediated bioremoval of organophosphorus pesticides using an indigenous Kosakonia oryzae strain -VITPSCQ3 in a Vertical-flow Packed Bed Biofilm Bioreactor.},
journal = {Ecotoxicology and environmental safety},
volume = {192},
number = {},
pages = {110290},
doi = {10.1016/j.ecoenv.2020.110290},
pmid = {32058164},
issn = {1090-2414},
abstract = {The widespread use of pesticides has been one of the major anthropogenic sources of environmental pollution. Organophosphorus (OP) pesticides are predominantly used in agriculture due to their broad-spectrum insecticidal activity and chemical stability. The study was focused on the biodegradation of OP pesticides, Profenofos (PF) and Quinalphos (QP) in culture media using bacterium isolated from wetland paddy rhizosphere. The strain VITPSCQ3 showed higher pesticide tolerance, efficient biofilm formation and was capable of synthesizing organophosphate degrading enzymes. Based on the 16S rRNA gene sequencing the isolate exhibited maximum sequence similarity with Kosakinia oryzae (GenBank accession number: KR149275). Biodegradation assay with various concentrations of PF and QP (200, 400, 600 and 800 mg L-1) showed maximum degradation up to 82% and 92% within 48 h. The kinetic studies revealed the biodegradation rates (k) to be 0.0844 min-1 and 0.107 min-1 with half-lives (h) of 18 h and 14.8 h for PF and QP. The degradation products were identified by GCMS and possible degradation pathways were proposed using Insilico techniques. To the best of our knowledge, this is the first report on the biodegradation of PF and QP using Kosakonia oryzae. Bioremoval of PF and QP from aqueous solution was performed using the biofilm of VITPSCQ3 developed on selected substrates in a circulating Vertical-flow packed-bed biofilm (VFPBB) bioreactor. Charcoal, gravel and mushroom (Agaricus bisporus) were used as biofilm carriers. Mushroom showed strong biofilm formation with optimum biodegradation capacity of up to 96% for PF and 92% for QP within 120 min reaction time.},
}

@article {pmid32057619,
year = {2020},
author = {Dey, P and Parai, D and Banerjee, M and Hossain, ST and Mukherjee, SK},
title = {Naringin sensitizes the antibiofilm effect of ciprofloxacin and tetracycline against Pseudomonas aeruginosa biofilm.},
journal = {International journal of medical microbiology : IJMM},
volume = {},
number = {},
pages = {151410},
doi = {10.1016/j.ijmm.2020.151410},
pmid = {32057619},
issn = {1618-0607},
abstract = {The study aims to explore the combinatorial effect of naringin with antibiotics, ciprofloxacin and tetracycline on Pseudomonas aeruginosa biofilms. The antibiofilm efficacy of selected treatment regimes against P. aeruginosa biofilm were quantified by crystal violet assay, MTT assay, Congo red binding assay, and were visualized by confocal laser scanning microscopy and scanning electron microscopy. All the assays reflected antibiofilm activities, however, combinatorial performances of naringin with antibiotics were found to be more significant. A significant reduction in swimming and swarming motilities along with pellicle formation and altered colony morphology were observed as a result of combinatorial effect. The cytotoxicity of naringin and its antibiotic combinations was assayed on murine macrophage cell line. The applicability of such combinations was tested for their relative eradication against pre-formed biofilm on urinary catheter surface. This finding indicated that naringin potentiates the efficacy of both ciprofloxacin and tetracycline on P. aeruginosa biofilm in comparison to their solo treatment. The finding would help to open hitherto unexplored possibilities of establishing naringin as a potential antibiofilm agent and suggest on the possibility of its use in drug-herb combinations for managing biofilm-associated bacterial infections.},
}

@article {pmid32056620,
year = {2020},
author = {Sikder, MNA and Xu, G and Xu, H},
title = {Seasonal variability in taxonomic breadth of biofilm-dwelling ciliates in colonization surveys for marine bioassessment.},
journal = {Marine pollution bulletin},
volume = {151},
number = {},
pages = {110828},
doi = {10.1016/j.marpolbul.2019.110828},
pmid = {32056620},
issn = {1879-3363},
abstract = {To determine an optimal sampling strategy for collecting samples with an expected taxonomic breadth, a 1-year baseline colonization survey was conducted in Chinese coastal waters using glass slides as an artificial substratum for biofilm-dwelling ciliates. A total of 240 slide samples were collected at a depth of 1 m in a four-season cycle. The taxonomic composition and structure of the ciliate communities differed from spring to winter. The colonization dynamics in taxonomic distinctness showed a significant variability among the four seasons. Expectation tests on the pairs of average taxonomic distinctness indices demonstrated a seasonal variability in taxonomic breadth of the ciliates, with high expectation levels in spring and autumn and low levels in the other two seasons. These findings suggest that there was a significant seasonal variability in taxonomic breadth for colonization surveys of biofilm-dwelling ciliates, and that an optimal sampling strategy should be determined for bioassessment in marine ecosystems.},
}

@article {pmid32056409,
year = {2020},
author = {Panariello, BH and Azabi, AA and Mokeem, LS and AlMady, FA and Lippert, F and Hara, AT and Duarte, S},
title = {The effects of charcoal dentifrices on Streptococcus mutans biofilm development and enamel demineralization.},
journal = {American journal of dentistry},
volume = {33},
number = {1},
pages = {12-16},
pmid = {32056409},
issn = {0894-8275},
abstract = {PURPOSE: To evaluate the in vitro effects of commercially available charcoal dentifrices on Streptococcus mutans biofilm development and their ability to prevent enamel demineralization.

METHODS: Streptococcus mutans biofilm was formed on polished bovine enamel specimens (n= 9 per treatment), and treated twice-daily for 120 seconds over the course of 5 days with: charcoal dentifrice containing fluoride (1,000 ppm F) (CF+), fluoride-free charcoal dentifrice (CF-), regular fluoride (1,100 ppm F) dentifrice (F+), or regular fluoride-free dentifrice (F-). Chlorhexidine (CHX, 0.12%) and deionized water (DIW) were used as positive and negative controls, respectively. Biofilms were analyzed for bacterial viability (colony-forming units, CFU). The pH of the medium was measured daily. Enamel specimens were analyzed using Vickers microhardness (HV) and transversal microradiography (TMR). Data were analyzed using one-way ANOVA followed by post-hoc tests (α= 0.05).

RESULTS: F+ showed higher pH values than CF+ and CF-, and CF- presented higher pH than CF+, showing that CF+ did not have inhibitory effects on the acidogenicity of cariogenic biofilms. CFU was significantly decreased when specimens were treated with CF+, CF- and F+, compared to specimens treated with DIW (P≤ 0.035) or F- (P≤ 0.001), respectively. However, the reduction observed was minimal (approximately 1 log). CF+ and CF- were less effective than F+ in preventing enamel demineralization as determined using HV (P= 0.041 and P= 0.003, respectively) and TMR (P≤ 0.001). Both charcoal dentifrices (CF+, CF-) did not show relevant inhibition of S. mutans biofilm growth. Additionally, neither product prevented enamel demineralization compared to a regular fluoride-containing dentifrice.

CLINICAL SIGNIFICANCE: The tested charcoal dentifrices did not exhibit anticaries potential.},
}

@article {pmid32055912,
year = {2020},
author = {Gao, XY and Liu, Y and Miao, LL and Liu, ZP},
title = {Pseudomonas sp. AOB-7 utilizes PHA granules as a sustained-release carbon source and biofilm carrier for aerobic denitrification of aquaculture water.},
journal = {Applied microbiology and biotechnology},
volume = {},
number = {},
pages = {},
doi = {10.1007/s00253-020-10452-y},
pmid = {32055912},
issn = {1432-0614},
support = {KFJ-BRP-009//Chinese Academy of Sciences/ ; },
abstract = {Nitrate accumulation causes long-time threat to aquatic animals in recirculating aquaculture system (RAS); thus, nitrate removal is also required in RASs. However, the lack of carbon sources makes denitrification difficult to function. Nitrate removal performance of an aerobic denitrifying and extracellular polyhydroxyalkanoate depolymerase-producing bacterium, Pseudomonas sp. AOB-7, using polyhydroxyalkanoate (PHA) granules as a solid sustained-release carbon source in RAS was evaluated. With the initial nitrate-N concentration of 140 mg/L, the high denitrification rates of 0.056 g NO3--N L-1 day-1 and 0.035 g NO3--N L-1 day-1 were achieved in denitrification medium containing poly-β-hydroxybutyrate (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), respectively. Significant erosions and pits formed on the surface of the granules made them a good biofilm carrier for AOB-7, and 3-hydroxybutyrate (3-HB) monomer was the major product released to aquatic phase, which was benefit to animals. SEM photos showed that AOB-7 entered and attached on the inside of the PHA particle holes. A 4-week application trial was conducted to reveal the effects of PHB (AOB-7) denitrifying agent and 3-HB produced on growth of zebrafish (Brachydanio rerio) by adding 0.1% (w/v) PHB (AOB-7) denitrifying agent. Result indicated that PHB (AOB-7) denitrifying agent can significantly reduce nitrate-N content in RASs. Compared with the control group, feed coefficient ratio reduced by 18% and weight gain ratio increased by 29% in the PHB (AOB-7) denitrifying agent group. 3-HB monomer produced during the denitrification was speculated to function as a prebiotic and promote zebrafish growth. KEY POINTS: • AOB-7 showed a good aerobic denitrifying ability on PHA granules as sustained-release C source. • PHB (AOB-7) denitrifying agent can significantly reduce nitrate content in RAS. • R-3-HB monomer was the major product released to aquatic phase and function as a prebiotic.},
}

@article {pmid32053733,
year = {2020},
author = {Yang, SY and Liu, Y and Mao, J and Wu, YB and Deng, YL and Qi, SC and Zhou, YC and Gong, SQ},
title = {The anti-biofilm and collagen-stabilizing effects of proanthocyanidin as an auxiliary endodontic irrigant.},
journal = {International endodontic journal},
volume = {},
number = {},
pages = {},
doi = {10.1111/iej.13280},
pmid = {32053733},
issn = {1365-2591},
abstract = {AIM: To evaluate the anti-biofilm effect of Proanthocyanidin (PA) solution as an irrigant against Enterococcus faecalis (E. faecalis) and its influence on the mechanical properties and biodegradation resistance of demineralized root dentine.

METHODOLOGY: E. faecalis were introduced into human root dentine tubules by a serial centrifugation method and grown for 1 week. Dentine blocks infected with 1-week-old E. faecalis biofilms were treated with the following irrigants: sterile water (control), 2% chlorhexidine (CHX), 2% PA, 5% PA and 10% PA. After treatment, the live and dead bacteria proportions within E. faecalis biofilms were analysed using confocal laser scanning microscopy. To evaluate the biostability of fully demineralized dentine treated by the aforementioned irrigants, the elastic modulus and hydroxyproline release of human dentine incubated in collagenase solution were tested at baseline, after irrigant treatment and after biodegradation, respectively. Furthermore, the surface chemical bond of demineralized dentine collagen treated by different irrigants was characterized by X-ray photoelectron spectroscopy (XPS). Statistical analysis was performed using one-way ANOVA and Tukey's post hoc multiple comparisons with the significance level at 5%.

RESULTS: The proportion of dead E. faecalis volume was significantly higher in the PA and CHX groups than that in the control group (p < 0.05). PA irrigation significantly increased the mechanical properties of demineralized dentine (p < 0.05) and the effect was enhanced with increasing PA concentration. CHX and PA groups had significantly less elasticity loss and hydroxyproline release (p < 0.05). The biomodification of dentine collagen by PA was verified by increased C-O/C-N peak percentage under C1s and C-O peak percentage under O1s narrow-scan XPS spectra.

CONCLUSIONS: PA killed E. faecalis within biofilms and enhanced the biostability of the collagen matrix of demineralized root dentine. It might be used as an auxiliary endodontic irrigant with anti-biofilm and collagen-stabilizing effects.},
}

@article {pmid32051871,
year = {2020},
author = {Manilal, A and Sabu, KR and Shewangizaw, M and Aklilu, A and Seid, M and Merdikios, B and Tsegaye, B},
title = {In vitro antibacterial activity of medicinal plants against biofilm-forming methicillin-resistant Staphylococcus aureus: efficacy of Moringa stenopetala and Rosmarinus officinalis extracts.},
journal = {Heliyon},
volume = {6},
number = {1},
pages = {e03303},
doi = {10.1016/j.heliyon.2020.e03303},
pmid = {32051871},
issn = {2405-8440},
abstract = {The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) is slowly rising in Ethiopia for the past few decades. Therefore, novel classes of antibiotics are indispensable to combat the increased incidence of newly emerging multidrug-resistant bacteria like MRSA. Terrestrial flora is considered as a reservoir of novel bioactive secondary metabolites as they have provided us with the largest array of natural products. In this background, the present study is intended to evaluate the in-vitro antibacterial efficacy of five medicinal plants (Ocimum lamiifolium Hochst. ex Benth., Rosmarinus officinalis L, Catharanthus roseus Linn., Azadirachta indica A. Juss and Moringa stenopetala Bac) against a panel of seven biofilm-forming MRSA. The leaves of the plants were extracted in organic solvents of varying polarity and the resultant crude extracts of respective medicinal plants were inspected for their antimicrobial activity by well diffusion technique. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the plant extracts against MRSA were determined by the broth dilution method. Besides, an anti-biofilm assay of the most potent plant extract was also performed, after which its chemical constituents were delineated by combined Gas Chromatographic and Mass Spectroscopic profiling (GC-MS). The results revealed that, of the five plants, three species including M. stenopetala, R. officinalis, and O. lamifolium exhibited significant antibacterial activity. Organic solvents with high and medium polarity were excellent in extracting antimicrobials compared to nonpolar solvents. The broadest and highest rank of activity was observed in the crude ethanolic extract of M. stenopetala. Based on the MIC/MBC ratio, the crude ethanolic extract of M. stenopetala was determined to be bacteriostatic. Anti-biofilm assay showed that the extract of M. stenopetala fairly inhibited the growth of MRSA in the preformed biofilm matrix. The GC-MS analysis of M. stenopetala revealed the presence of twelve compounds with antimicrobial activity. The present study provides new insight into the development of novel drug leads to the management of MRSA.},
}

@article {pmid32051263,
year = {2020},
author = {Harro, JM and Shirtliff, ME and Arnold, W and Kofonow, JM and Dammling, C and Achermann, Y and Brao, K and Parvizi, J and Leid, JG},
title = {Development of a Novel and Rapid Antibody-based Diagnostic for Chronic Staphylococcus aureus Infections Based on Biofilm Antigens.},
journal = {Journal of clinical microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1128/JCM.01414-19},
pmid = {32051263},
issn = {1098-660X},
abstract = {Prosthetic joint infections are difficult to diagnose and treat due to biofilm formation by the causative pathogens. Pathogen identification relies on microbial culture that requires days-to-weeks, and in the case of chronic biofilm infections, lacks sensitivity. Diagnosis of infection is often delayed past the point of effective treatment such that only the removal of the implant is curative. Early diagnosis of an infection based on antibody detection might lead to less invasive, early interventions. Our study examined antibody-based assays against the Staphylococcus aureus biofilm-upregulated antigens SAOCOL0486 (a lipoprotein), glucosaminidase (a domain of SACOL1062), and SACOL0688 (the manganese transporter MntC) for detection of chronic S. aureus infection. We evaluated these antigens by enzyme-linked immunosorbent assay (ELISA) using sera from naive rabbits and rabbits with S. aureus-mediated osteomyelitis, and then validated a proof of concept for the lateral flow assay (LFA). The SACOL0688 LFA demonstrated 100% specificity and 100% sensitivity. We demonstrated the clinical diagnostic utility of the SACOL0688 antigen using synovial fluid (SF) from humans with orthopedic implant infections. Elevated antibody levels to SACOL0688 in clinical SF specimens correlated to 91% sensitivity and 100% specificity for the diagnosis of S. aureus infection by ELISA. We found measuring antibodies levels to SACOL0688 in SF using ELISA or LFA provides a tool for the sensitive and specific diagnosis of S. aureus prosthetic joint infection. Development of the LFA diagnostic modality is a desirable, cost-effective option, potentially providing rapid readout in minutes for chronic biofilm infections.},
}

@article {pmid32050878,
year = {2020},
author = {Zeybek, Z and Türkmen, A},
title = {[Investigation of the Incidence of Legionella and Free-Living Amoebae in Swimming Pool Waters and Biofilm Specimens in Istanbul by Different Methods].},
journal = {Mikrobiyoloji bulteni},
volume = {54},
number = {1},
pages = {50-65},
doi = {10.5578/mb.68962},
pmid = {32050878},
issn = {0374-9096},
abstract = {Legionella bacteria living in free form or in biofilm and free-living amoebae (FLA) can infect humans through swimming pools and can cause various diseases. FLA may also threaten the health of swimmers because they are capable of being hosts for Legionella and some other bacteria. The aim of this study was to investigate the presence of total aerobic heterotrophic bacteria (TAHB), FLA and Legionella bacteria in swimming pool waters and biofilm samples in Istanbul by using culture and FISH methods. Water plate count agar (wPCA), buffered charcoal yeast extract (BCYE) agar supplemented with glycinevancomycin-polymyxin-cycloheximide (GVPC) and Escherichia coli cultivated non-nutrient agar (NNA) were used for the culture of TAHB, Legionella and FLA. For the FISH method analysis , Leg 705 and Leg PNE1 probes labeled with fluorescent dye for Legionella and ACANTHA probe for Acanthamoeba genus FLA were used. Legionella pneumophila serogroup 1 ATCC 33152, L.pneumophila serogroup 3 ATCC 33155 and Acanthamoeba castellani ATCC 50373 were used as positive controls. TAHB were grown in 92% and 84% of water and biofilm samples. Although Legionella bacteria could not be grown in any of the water samples, it was detected in 6 (24%) water samples by FISH method. Although these bacteria could be grown in 1 (4%) of biofilm samples, 7 (28%) were detected by FISH method. FLA were found to be 16% by culture in water samples and 28% by FISH analysis. These amoebae were detected 8% and 20% in biofilm samples by culture and FISH method, respectively. It was determined that one of the isolates of FLA had thermotolerant activity (potentially pathogenic). L.pneumophila serogroup 1 was detected in one water sample and in four biofilm samples. According to the culture method, TAHB and FLA were found to be more common in water samples than in biofilm samples and Legionella bacteria were more common in biofilm samples than in water samples (p≤ 0.05). In the detection of Legionella bacteria, the superiority of FISH method compared to culture method was found to be statistically significant (p≤ 0.05). In this study, it was found that the number of TAHB in the controlled swimming pools was within the limits determined by the Ministry of Health (≤ 200 cfu/ml). It will be appropriate to examine both water and biofilm samples for the investigation of TAHB, FLA and Legionella. It may be appropriate to use both culture and FISH methods to detect the presence of FLA in water and biofilm samples. This study is the first study to investigate the presence of Legionella and FLA in swimming pools in Istanbul, and further studies are needed to examine more pool water and biofilm samples. With the data obtained, the health principles and controls of swimming pools will be re-considered and will be contributed to public health.},
}

@article {pmid32050877,
year = {2020},
author = {Altınok, Ö and Boral, B and Ergin, A and Köseoğlu Eser, Ö},
title = {[Existence of Biofilm and Biofilm-Associated Virulence Genes in Multi-Drug Resistant Invasive Acinetobacter baumannii Isolates].},
journal = {Mikrobiyoloji bulteni},
volume = {54},
number = {1},
pages = {40-49},
doi = {10.5578/mb.20204},
pmid = {32050877},
issn = {0374-9096},
abstract = {Acinetobacter baumannii is a multi-drug resistant (MDR) gram-negative pathogen leading to nosocomial infections. Hospital-acquired infections due to A.baumannii occur especially in patients hospitalized in intensive care units. Important infections related to this bacterium are pneumonia, bacteremia, endocarditis, skin and soft tissue, urinary tract infections and meningitis. Human transmission is usually through the hospital environment or through medical personnel. A.baumannii isolates increases their virulence not only being multiple resistance to antibiotics but as well as the ability to form biofilm. The biofilm formation of A.baumannii isolates were mostly related with genes encoding curli fiber (csgA), the chaperone-usher fimbria (csuE) and the outer membrane (ompA). The aim of this study was to demonstrate biofilm production and virulence genes in MDR invasive A.baumannii isolates. MDR and similarity status previously known invasive A.baumannii (n= 156) isolates were included in the study. Biofilm production was determined by quantitative microplate biofilm method. Virulence genes csgA, csuE, fimH, ompA and blaPER-1 were investigated by polymerase chain reaction (PCR). It was determined that 60.3% (94/156) of all the isolates formed biofilm. Of these 94 isolates, 17 were weak, 33 were medium and 44 were strong. The mean biomass forming capacity of the isolates was found to be 2.23 ± 0.0033. Among the isolates included in the study (n= 156) the frequency of csgA, csuE, ompA, fimH and blaPER-1 genes were 71.2%, 32.1%, 21.8%, 7.1% and 3.2% respectively. The frequency of csgA, ompA, bap, csuE, fimH virulence genes were found to be 41.5%, 24.5%, 20.2% and 5.3% among biofilm positive isolates respectively. Biofilm-forming isolates were most commonly found in pulsotype II 19.1% (18/94), pulsotype IX 17.0% (16/94) and pulsotype VI 12.8% (12/94). In this study, when the distribution of virulence genes were compAred with the isolates that have weak, medium and strong biofilm, all of the studied genes were found to be more abundant in isolates with strong and medium positive biofilm production. This has shown that excluding fimH gene, csgA, csuE and ompA genes have contributed to the biofilm formation in invasive A.baumannii isolates, respectively.},
}

@article {pmid32050341,
year = {2020},
author = {Huang, ZS and Wei, ZS and Xiao, XL and Li, BL and Ming, S and Cheng, XL and Jiao, HY},
title = {Bioconversion of Hg0 into HA-Hg for simultaneous removal of Hg0 and NO in a denitrifying membrane biofilm reactor.},
journal = {Chemosphere},
volume = {244},
number = {},
pages = {125544},
doi = {10.1016/j.chemosphere.2019.125544},
pmid = {32050341},
issn = {1879-1298},
abstract = {Bacterial mercury oxidation coupled to denitrification offers great potential for simultaneous removal of elemental mercury (Hg0) and nitric oxide (NO) in a denitrifying membrane biofilm reactor (MBfR). Four potentially contributory mechanisms tested separately, namely, membrane gas separation, medium absorption, biosorption and biotransformation, which contributed 4.9%/7.2%, 8.1%/8.9%, 38.8%/9.5% and 48.2%/84.9% of overall Hg0/NO removal in MBfR. Herein, Hg0 bio-oxidation, oxidative Hg0 biosorption and denitrification played leading roles in simultaneous removal of Hg0 and NO. Living microbes performed simultaneous Hg0 bio-oxidation and denitrification, in which Hg0 as electron donor was biologically oxidized to oxidized mercury (Hg2+), while NO as the terminal electron acceptor was denitrified to N2. The Hg2+ further complexed with humic acids in extracellular polymeric substances via functional groups (-SH, -OH, -NH- and -COO-) and formed humic acids bound mercury (HA-Hg). Non-living microbial matrix performed oxidative Hg0 biosorption, in which Hg0 may be physically adsorbed by cellular matrix, then non-metabolically oxidized to Hg2+ via oxidative complexation with -SH in humic acids and finally cleavage of S-H bond and surface charge transfer led to formation of HA-Hg. Therefore, bioconversion of Hg0 to HA-Hg by Hg0 bio-oxidation and oxidative Hg0 biosorption coupled with NO denitrification to N2 dynamically cooperated to accomplish simultaneous removal of Hg0 and NO in MBfR.},
}

@article {pmid32049553,
year = {2020},
author = {Jacquier, H and Vironneau, P and Dang, H and Verillaud, B and Lamers, G and Herman, P and Vicaut, E and Tessier, N and Bidet, P and Varon, E and Van Den Abbeele, T and Cambau, E and Bercot, B and Kania, R},
title = {Bacterial biofilm in adenoids of children with chronic otitis media. Part II: a case-control study of nasopharyngeal microbiota, virulence, and resistance of biofilms in adenoids.},
journal = {Acta oto-laryngologica},
volume = {},
number = {},
pages = {1-5},
doi = {10.1080/00016489.2020.1718749},
pmid = {32049553},
issn = {1651-2251},
abstract = {Background: We previously described that adenoid tissue in children with chronic otitis media (COM) contained more mucosal biofilms than adenoid tissue removed for hypertrophy.Aims/objectives: The aim of the second part was to characterize nasopharyngeal microbiota and explore virulence of the most common middle ear pathogens.Material and methods: Bacteriological analysis was performed following a culture-based approach on the samples recovered from 30 patients of COM group (15 biofilm-positive and 15 biofilm-negative) and from 30 patients of a control group (15 biofilm-positive and 15 biofilm-negative). Virulence factors of Streptococcus pneumoniae, Streptococcus pyogenes, and Haemophilus influenzae were investigated.Results: The most frequent species were Firmicutes followed by Proteobacteria and Actinobacteria. The presence of biofilm was statistically associated with an increase of the number of bacterial species and Firmicutes phylum regardless of the condition (case/control). No virulence factors associated with invasive isolates were found for the most common middle ear pathogens.Conclusions and significance: This case-control study demonstrated that the presence of COM plus biofilm was associated with a given microbiota which contained more Firmicutes. Our study allows a better understanding of physiopathological mechanisms involved in chronic otitis media and paves the way for further investigations.},
}

@article {pmid32048958,
year = {2020},
author = {Janga, YS and Mosolygó, T},
title = {Inhibition of Bacterial Biofilm Formation by Phytotherapeutics with Focus on Overcoming Antimicrobial Resistance.},
journal = {Current pharmaceutical design},
volume = {},
number = {},
pages = {},
doi = {10.2174/1381612826666200212121710},
pmid = {32048958},
issn = {1873-4286},
abstract = {Bacteria within biofilms are more resistant to antibiotics and chemical agents than planktonic bacteria in suspension. Treatment of biofilm-associated infections inevitably involves high dosages and prolonged courses of antimicrobial agents; therefore, there is a potential risk of the development of antimicrobial resistance (AMR). Due to the high prevalence of AMR and its association with biofilm formation, investigation of more effective anti-biofilm agents is required. From ancient times, herbs and spices have been used to preserve foods, and their antimicrobial, anti-biofilm and anti-quorum sensing properties are well known. Moreover, phytochemicals exert their anti-biofilm properties at sub-inhibitory concentrations without providing the opportunity for the emergence of resistant bacteria or harming the host microbiota. With increasing scientific attention to natural phytotherapeutic agents, numerous experimental investigations have been conducted in recent years. The present paper aims to review the articles published in the last decade in order to summarize a) our current understanding of AMR in correlation with biofilm formation and b) the evidence of phytotherapeutic agents against bacterial biofilms and their mechanisms of action. The main focus has been put on herbal anti-biofilm compounds tested to date in association with Staphylococcus aureus, Pseudomonas aeruginosa and food-borne pathogens (Salmonella spp., Campylobacter spp., Listeria monocytogenes and Escherichia coli).},
}

@article {pmid32048076,
year = {2020},
author = {Fu, H and Chen, F and Liu, W and Kong, W and Wang, C and Fang, X and Ye, J},
title = {Adding nutrients to the biocontrol strain JK-SH007 promotes biofilm formation and improves resistance to stress.},
journal = {AMB Express},
volume = {10},
number = {1},
pages = {32},
doi = {10.1186/s13568-019-0929-8},
pmid = {32048076},
issn = {2191-0855},
support = {CX [16] 1005//the agricultural Independent Innovation Project of Jiangsu Province/ ; },
abstract = {Burkholderia pyrrocinia JK-SH007 is an important biocontrol strain for the prevention and treatment of poplar canker disease. Its powerful biocontrol function is inseparable from its successful colonization of poplar trees. Bacterial biofilms can ensure the long-term colonization of a host. To explore the mechanism of action of biofilms in the biocontrol process, we manipulated various exogenous factors to explore the morphology of the JK-SH007 biofilm in vitro. The addition of glycerol and MgSO4 to TSB medium stimulated biofilm production, increased the resistance of JK-SH007 to disease, enhanced the survival of JK-SH007 in nutrient-poor environments and maintained the antagonistic ability of JK-SH007 against the poplar canker pathogen. Therefore, we constructed and optimized a biofilm-forming system to produce a large number of stable JK-SH007 biofilms. The optimized system showed that the optimal incubation time for JK-SH007 biofilm formation was 14 h, the optimal temperature of the static culture was 25 °C, and the optimal pH was 5. The optimal medium for biofilm formation was TSB medium, 1% glycerol and 50 mM MgSO4. RT-qPCR experiments showed that an increase in the expression of the suhB gene promoted JK-SH007 biofilm formation, while an increase in the expression level of the ropN gene inhibited JK-SH007 biofilm formation. The possible mechanism by which JK-SH007 was inhibited by biofilm formation under natural culture was revealed. These results indicate the importance of adding nutrients to JK-SH007 biocides produced on a commercial scale. This is the first report of JK-SH007 producing a long-lasting biofilm that guarantees antagonism.},
}

@article {pmid32048056,
year = {2020},
author = {Pedroza-Dávila, U and Uribe-Alvarez, C and Morales-García, L and Espinoza-Simón, E and Méndez-Romero, O and Muhlia-Almazán, A and Chiquete-Félix, N and Uribe-Carvajal, S},
title = {Metabolism, ATP production and biofilm generation by Staphylococcus epidermidis in either respiratory or fermentative conditions.},
journal = {AMB Express},
volume = {10},
number = {1},
pages = {31},
doi = {10.1186/s13568-020-00966-z},
pmid = {32048056},
issn = {2191-0855},
support = {IN203018//UNAM DGAPA PAPIIT/ ; },
abstract = {Staphylococcus epidermidis is a Gram-positive saprophytic bacterium found in the microaerobic/anaerobic layers of the skin that becomes a health hazard when it is carried across the skin through punctures or wounds. Pathogenicity is enhanced by the ability of S. epidermidis to associate into biofilms, where it avoids attacks by the host and antibiotics. To test the effect of oxygen on metabolism and biofilm generation, cells were cultured at different oxygen concentrations ([O2]). As [O2] decreased, S. epidermidis metabolism went from respiratory to fermentative. Remarkably, the rate of growth decreased at low [O2] while a high concentration of ATP ([ATP]) was kept. Under hypoxic conditions bacteria associated into biofilms. Aerobic activity sensitized the cell to hydrogen peroxide-mediated damage. In the presence of metabolic inhibitors, biofilm formation decreased. It is suggested that at low [O2] S. epidermidis limits its growth and develops the ability to form biofilms.},
}

@article {pmid32046702,
year = {2020},
author = {Alshahrani, AM and Gregory, RL},
title = {In vitro Cariostatic effects of cinnamon water extract on nicotine-induced Streptococcus mutans biofilm.},
journal = {BMC complementary medicine and therapies},
volume = {20},
number = {1},
pages = {45},
doi = {10.1186/s12906-020-2840-x},
pmid = {32046702},
issn = {2662-7671},
abstract = {BACKGROUND: Dental caries is one of the most prevalent chronic oral diseases worldwide. Dental caries is mainly associated with Streptococcus mutans and the Lactobacillus species. A specific relationship was found between nicotine and S. mutans growth as the presence of nicotine increased S. mutans biofilm formation. Nicotine is able to increase the number of S. mutans and extracellular polysaccharide (EPS) synthesis. Among the widely used herbs and spices is cinnamon which demonstrated a strong antibacterial activity against a wide variety of bacteria including S. mutans and showed the ability to inhibit S. mutans biofilm formation. Cinnamon essential oil, obtained from the leaves of C. zeylanicum, has been demonstrated to be effective against S. mutans and Lactobacillus acidophilus, which are partially responsible for dental plaque formation and caries development. The aim of this study was to identify the effects of nicotine exposure on the inhibitory effects of cinnamon water extract on S. mutans biofilm formation.

MATERIALS AND METHODS: A 24-h culture of S. mutans UA159 in microtiter plates was treated with varying nicotine concentrations (0-32 mg/ml) in Tryptic Soy broth supplemented with 1% sucrose (TSBS) with or without a standardized concentration (2.5 mg/ml) of cinnamon water extract. A spectrophotometer was used to determine total growth absorbance and planktonic growth. The microtiter plate wells were washed, fixed and stained with crystal violet dye and the absorbance measured to determine biofilm formation.

RESULTS: The presence of 2.5 mg/ml cinnamon water extract inhibits nicotine-induced S. mutans biofilm formation from 34 to 98% at different concentrations of nicotine (0-32 mg/ml).

CONCLUSION: The results demonstrated nicotine-induced S. mutans biofilm formation is decreased from 34 to 98% in the presence of 2.5 mg/ml cinnamon water extract. This provides further evidence about the biofilm inhibitory properties of cinnamon water extract and reconfirms the harmful effects of nicotine.},
}

@article {pmid32045645,
year = {2020},
author = {Mirzaei, R and Mirzaei, H and Alikhani, MY and Sholeh, M and Arabestani, MR and Saeedijam, M and Karampoor, S and Ahmadyousefi, Y and Moghadam, MS and Irajian, GR and Hasanvand, H and Yousefimashouf, R},
title = {Bacterial biofilm in colorectal cancer: What is the real mechanism of action?.},
journal = {Microbial pathogenesis},
volume = {},
number = {},
pages = {104052},
doi = {10.1016/j.micpath.2020.104052},
pmid = {32045645},
issn = {1096-1208},
abstract = {Human colorectal cancer is the third most common cancer around the world. Colorectal cancer has various risk factors, but current works have bolded a significant activity for the microbiota of the human colon in the development of this disease. Bacterial biofilm has been mediated to non-malignant pathologies like inflammatory bowel disease but has not been fully documented in the setting of colorectal cancer. The investigation has currently found that bacterial biofilm is mediated to colon cancer in the human and linked to the location of human cancer, with almost all right-sided adenomas of colon cancers possessing bacterial biofilm, whilst left-sided cancer is rarely biofilm positive. The profound comprehension of the changes in colorectal cancer can provide interesting novel concepts for anticancer treatments. In this review, we will summarize and examine the new knowledge about the links between colorectal cancer and bacterial biofilm.},
}

@article {pmid32044647,
year = {2020},
author = {Wang, S and Ma, L and Xu, Y and Wang, Y and Zhu, N and Liu, J and Dolfing, J and Kerr, P and Wu, Y},
title = {The unexpected concentration-dependent response of periphytic biofilm during indole acetic acid removal.},
journal = {Bioresource technology},
volume = {303},
number = {},
pages = {122922},
doi = {10.1016/j.biortech.2020.122922},
pmid = {32044647},
issn = {1873-2976},
abstract = {Due to its extensive application in agriculture as a germinating agent and growth promoter, indole acetic acid (IAA) is present in a variety of aquatic ecosystems. To explore the response of microbial aggregates to exogenous IAA in aquatic ecosystems, periphytic biofilm, a typical microbial aggregate, was exposed to IAA at different concentrations. Results reveal an unexpected concentration-dependent effect of IAA on periphytic biofilm. Concentrations of IAA less than 10 mg/L inhibit periphytic growth, but stimulate growth when the IAA concentration exceeds 50 mg/L. Periphytic biofilm adapts to different IAA concentrations by antioxidant enzyme activation, community structure optimization and carbon-metabolism pattern change, and promotes bioremediation of IAA contaminated water in the process. The removal rates of IAA reached up to 95%-100%. This study reveals the capacity of periphytic biofilm for IAA removal in practice.},
}

@article {pmid32043899,
year = {2020},
author = {Dávila-Aviña, J and Gil-Solís, C and Merino-Mascorro, J and García, S and Heredia, N},
title = {Phenolics with Bactericidal Activity Alter Motility and Biofilm Formation in Enterotoxigenic, Enteropathogenic, and Enterohemorrhagic Escherichia coli.},
journal = {Foodborne pathogens and disease},
volume = {},
number = {},
pages = {},
doi = {10.1089/fpd.2019.2766},
pmid = {32043899},
issn = {1556-7125},
abstract = {Most Escherichia coli strains are innocuous to human beings; however, some strains can cause diarrhea and are grouped into pathotypes. Since current trends promote the use of natural-origin compounds to control bacteria, in this study, the effects of the phenolic compounds (PCs) tannic acid (TA), gallic acid (GA), methyl gallate (MG), and epigallocatechin gallate (EG) on the growth, swarming motility, biofilm formation, and expression of selected virulence genes of three E. coli pathotypes (enteropathogenic Escherichia coli [EPEC], enterohemorrhagic Escherichia coli [EHEC], and enterotoxigenic Escherichia coli [ETEC]) were evaluated. Minimum bactericidal concentrations (MBCs) were determined by using microtiter plates, and the effects of sublethal PC concentrations on swarming motility were evaluated on Luria-Bertani agar. Biofilm formation was assessed in microtiter plates via crystal violet staining, and the expression levels of genes involved in biofilm formation (flhC, fliA, fliC, and csgA) and swarming motility (csgD and cyaA) were evaluated via quantitative PCR. All PC were bactericidal with minimal bactericidal concentrations ranging from 0.07 to 2.1 mg/mL. At concentrations lower than the MBC, PCs decreased swarming motility (14.8-100%). GA reduced biofilm formation in all of the tested strains; however, TA, MG, and EG induced biofilm formation in some strains at specific concentrations. TA induced the overexpression of csgA, csgD, and cyaA, whereas the other PCs did not have any effects or reduced their expression levels. The PCs tested in this study showed potential to control E. coli strains belonging to the EHEC, ETEC, and EPEC pathotypes by affecting their growth, swarming motility, and virulence gene expression; however, proper concentrations must be used to avoid the induction of undesirable virulence factor genes.},
}

@article {pmid32043412,
year = {2020},
author = {Attanasio, V and Di Luca, M and Carozza, A and Severino, S and Pallotto, C and Capoluongo, N and Palmiero, G and Bernardo, M and Tascini, C},
title = {Clinical efficacy of amoxicillin/clavulanate plus cefditoren as de-escalation combination therapy for endocarditis due to strongly biofilm-forming Enterococcus faecalis.},
journal = {Infectious diseases (London, England)},
volume = {},
number = {},
pages = {1-4},
doi = {10.1080/23744235.2020.1725109},
pmid = {32043412},
issn = {2374-4243},
}

@article {pmid32043363,
year = {2020},
author = {Bose, SK and Chauhan, M and Dhingra, N and Chhibber, S and Harjai, K},
title = {Terpinen-4-ol attenuates quorum sensing regulated virulence factors and biofilm formation in Pseudomonas aeruginosa.},
journal = {Future microbiology},
volume = {},
number = {},
pages = {},
doi = {10.2217/fmb-2019-0204},
pmid = {32043363},
issn = {1746-0921},
abstract = {Aim: To investigate the effects of Terpinen-4-ol on quorum sensing (QS)-regulated biofilm formation and virulence factors production in Pseudomonas aeruginosa. Materials & methods: QS inhibition, molecular docking analysis and gene expression studies were performed to check attenuation effect of Terpinen-4-ol on virulence of P. aeruginosa. Production of various virulence factors and biofilm formation were studied at sub-MIC of Terpinen-4-ol alone and in combination with ciprofloxacin. Results: Terpinen-4-ol at sub-MIC exhibited QS inhibition and downregulated all key QS genes. Molecular docking analysis showed high binding affinities of Terpinen-4-ol with QS receptors. Terpinen-4-ol exhibited synergistic interaction with ciprofloxacin and further reduced production of all the virulence factors and biofilms formation. Conclusion: Terpinen-4-ol could be developed into antivirulence drug after its in vivo evaluation for treatment strategies.},
}

@article {pmid32042258,
year = {2020},
author = {Bukhari, SI and Aleanizy, FS},
title = {Association of OprF mutant and disturbance of biofilm and pyocyanin virulence in pseudomonas aeruginosa.},
journal = {Saudi pharmaceutical journal : SPJ : the official publication of the Saudi Pharmaceutical Society},
volume = {28},
number = {2},
pages = {196-200},
doi = {10.1016/j.jsps.2019.11.021},
pmid = {32042258},
issn = {1319-0164},
abstract = {Outer membrane porin F (OprF) is a major structural membrane protein of Pseudomonas aeruginosa, a recognised human opportunistic pathogen which is correlated with severe hospital-acquired infections. This study investigating a multiphenotypic approach, based on the comparative study of a wild type strain of P. aeruginosa, its isogenic OprF mutant. Both P. aeruginosa PAO1 and OprF mutant strains were grown in same condition and cultures were subjected to further analysis by SDS PAGE, pyocyanin production and biofilm formation that was analyse using scanning electron microscopy. Based on biofilm formation essay and pyocyanin production, the study showed that OprF plays a dynamic role in P. aeruginosa virulence. The absence of OprF results in slow growth rate corresponded to elongated lag phase and reduced biofilm production also a significance reduction in the production of the quorum-sensing-dependent virulence factors pyocyanin. Accordingly, in the OprF mutant scanning electron microscope "SEM" images showed impaired cellular niche and detached cells when compared to regular attached P. aeruginosa wild type cells in the niche. Taken together, this study shows the contribution of OprF in P. aeruginosa virulence, at least partly through impairment of biofilm, cell to cell attachment in niche and pyocyanin production. This study show a vital link between OprF and virulence factor production, providing novel insights for its role in pathogenicity and future could provide the basis for the development of novel drug targets for antibiotics and vaccines.},
}

@article {pmid32042100,
year = {2020},
author = {Souza, JGS and Bertolini, M and Thompson, A and Mansfield, JM and Grassmann, AA and Maas, K and Caimano, MJ and Barao, VAR and Vickerman, MM and Dongari-Bagtzoglou, A},
title = {Role of glucosyltransferase R in biofilm interactions between Streptococcus oralis and Candida albicans.},
journal = {The ISME journal},
volume = {},
number = {},
pages = {},
doi = {10.1038/s41396-020-0608-4},
pmid = {32042100},
issn = {1751-7370},
support = {RO1 DE013986//U.S. Department of Health & Human Services | NIH | National Institute of Dental and Craniofacial Research (NIDCR)/ ; RO1AI029735//U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (NIAID)/ ; R21AI126146//U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (NIAID)/ ; },
abstract = {Streptococcal glucosyltransferases (Gtf) synthesize α-glucan exopolymers which contribute to biofilm matrix. Streptococcus oralis interacts with the opportunistic pathogen Candida albicans to form hypervirulent biofilms. S. oralis 34 has a single gtf gene (gtfR). However, the role of gtfR in single and mixed species biofilms with C. albicans has never been examined. A gtfR deletion mutant, purified GtfR, and recombinant GtfR glucan-binding domain were tested in single and mixed biofilms on different substrata in vitro. A mouse oral infection model was also used. We found that in single species biofilms growing with sucrose on abiotic surfaces S. oralis gtfR increased biofilm matrix, but not bacterial biomass. In biofilms with C. albicans, S. oralis encoding gtfR showed increased bacterial biomass on all surfaces. C. albicans had a positive effect on α-glucan synthesis, and α-glucans increased C. albicans accretion on abiotic surfaces. In single and mixed infection of mice receiving sucrose S. oralis gtfR enhanced mucosal burdens. However, sucrose had a negative impact on C. albicans burdens and reduced S. oralis burdens in co-infected mice. Our data provide new insights on the GtfR-mediated interactions between the two organisms and the influence of biofilm substratum and the mucosal environment on these interactions.},
}

@article {pmid32041788,
year = {2020},
author = {Gries, CM and Biddle, T and Bose, JL and Kielian, T and Lo, DD},
title = {Staphylococcus aureus fibronectin binding protein A mediates biofilm development and infection.},
journal = {Infection and immunity},
volume = {},
number = {},
pages = {},
doi = {10.1128/IAI.00859-19},
pmid = {32041788},
issn = {1098-5522},
abstract = {Implanted medical device-associated infections pose significant health risks as they are often the result of bacterial biofilm formation. Staphylococcus aureus is a leading cause of biofilm-associated infections which persist due to mechanisms of device surface adhesion, biofilm accumulation, and reprogramming of host innate immune responses. We found that the S. aureus fibronectin binding protein A (FnBPA) is required for normal biofilm development in mammalian serum, and that the SaeRS two-component system is required for functional FnBPA activity in serum. Furthermore, serum-developed biofilms deficient in FnBPA were more susceptible to macrophage invasion, and in a model of biofilm-associated implant infection, we found that FnBPA is crucial for the establishment of infection. Together, these findings show that S. aureus FnBPA plays an important role in physical biofilm development and represents potential therapeutic target for the prevention and treatment of device-associated infections.},
}

@article {pmid32040901,
year = {2019},
author = {Godovalov, AP and Stepanov, MS and Yakovlev, MV and Kobzarenko, EE and Batog, KA},
title = {[Determination of biofilm forming activity of microorganisms on synthetic polymeric materials.].},
journal = {Klinicheskaia laboratornaia diagnostika},
volume = {64},
number = {12},
pages = {758-761},
doi = {10.18821/0869-2084-2019-64-12-758-761},
pmid = {32040901},
issn = {0869-2084},
abstract = {Microorganisms are able to form biofilms on surfaces of biotic and abiotic nature. In turn, in human biotopes there are optimal conditions for the implementation of biofilm-forming activity. Moreover, in medical practice, polymeric materials are often used for drainage or prosthetics, which can also be successfully colonized by bacteria. However, in laboratory practice, the formation of biofilms is usually evaluated on glass or polystyrene. The purpose of the study is to evaluate the methodological features of studying the biofilm-forming activity of microorganisms on the surface of synthetic polymeric materials. We used strains of Staphylococcus aureus ATCC 25923, Escherichia coli K-12, Candida albicans ATCC 10231, as well as synthetic polymeric materials - DentLight Flow light-curing composite material (nano-hybrid fluid composite; Russia), glass ionomer chemical curing Fuji 1 (Japan), cement for temporary fixation of orthopedic constructions TempBond NE (USA), acrylic, polyurethane and polyvinyl chloride. The formation of biofilms in flat-bottomed ELISA plates in this study was considered as a control group. If the polymer belonged to cold curing materials, sterile flat-bottomed tablets were used, the bottom of which was filled with a thin layer of plastic. After hardening of the plastic, biofilms were formed in the tablets. In the second series of experiments, hot cured materials cut into equal parts 5×5×1 mm in size were placed in the wells of a plate and again used to determine biofilm formation with subsequent coloring. To extract the dye, the pieces were transferred to a new plate to exclude the amount of film biomass formed on the walls of the plate wells. In both cases, cultivation was carried out at 37° C for 24-48 hours. The biomass of the film was stained with fuchsin. Statistical data processing was performed using t-Student criterion. For the threshold level of significance, the value p <0.05 was taken. It is established that the proposed options for determining biofilm forming ability are available and indicative. It was revealed that the same microorganisms have individual biofilm formation indicators for each polymer material. The light curing dental composite and polyvinyl chloride exhibit the more pronounced antiadhesive properties than cements and polyurethane. Up to date, most of the studies of biofilm formation have been carried out using glass or polystyrene, which, as a rule, are not used for the manufacture of prostheses, catheters, drains, etc., which makes it difficult to assess the true film-forming activity of microorganisms. The proposed methodological approaches, especially the second option for preparing testing samples, solve this problem. In general, the proposed approaches to testing biofilm-forming activity on polymers are very simple to implement and generally available. For an adequate study of the biofilms formation, it will be advisable to use polymer materials, directly used in medicine, rather than polystyrene tablets, the material of which is found exclusively in laboratory practice.},
}

@article {pmid32039189,
year = {2020},
author = {Hoseinzadeh, E and Wei, C and Farzadkia, M and Rezaee, A},
title = {Effects of Low Frequency-Low Voltage Alternating Electric Current on Apoptosis Progression in Bioelectrical Reactor Biofilm.},
journal = {Frontiers in bioengineering and biotechnology},
volume = {8},
number = {},
pages = {2},
doi = {10.3389/fbioe.2020.00002},
pmid = {32039189},
issn = {2296-4185},
abstract = {Bioelectrochemical systems have undergone several modifications to promote the enzymes or pathways used to reduce the energy required for microbial metabolism. Changes in dominant bacteria, population, and growth rates occur when an electric current is applied intermittently. Applying electricity to bioelectrical reactor (BER) biofilms can either stimulate cells or lead to cell death; therefore, determining the applied voltage range that leads to viable and stimulated bacteria is crucial. We investigated the progression of apoptosis induced by a low frequency-low voltage alternating electric current (AC) in a BER biofilm and found that biofilms on carbon cloth (CC) and stainless steel (SS) 304 electrodes had pHzpc values of 8.67. The pHzpc of the biofilms increased by two compared to that of the inoculant bacteria mass. Furthermore, the Henderson-Hasselbalch equation reveals that the compositions of cell walls of the biofilms that formed on the CC and SS304 electrodes are very similar. In contrast, the CC and SS304 biofilms differ from the inoculant biomass without the influence of an AC field; this indicates that there are differences in the compositions of the cell walls in the present bacteria. Fourier transform infrared spectroscopy was used to compare spectra of the biofilms with that of the inoculation mass, and there were differences in shape and absorbance intensity, indicating variability in the composition, and quantity of each individual biofilm component. In addition, the dehydrogenase activity (DHA) content varied under different applied voltages; the highest DHA was obtained at 8 Vpp. A flow cytometry analysis showed a relatively low number of apoptotic cells (10.93 ± 5.19%) for the AC amplitudes studied. Thus, a low voltage-low frequency AC likely induces significant changes in bacterial metabolic activity but causes no significant change in their viability.},
}

@article {pmid32039129,
year = {2019},
author = {Stapleton, EM and Manges, R and Parker, G and Stone, EA and Peters, TM and Blount, RJ and Noriega, J and Li, X and Zabner, J and Polgreen, PM and Chipara, O and Herman, T and Comellas, AP},
title = {Indoor Particulate Matter From Smoker Homes Induces Bacterial Growth, Biofilm Formation, and Impairs Airway Antimicrobial Activity. A Pilot Study.},
journal = {Frontiers in public health},
volume = {7},
number = {},
pages = {418},
doi = {10.3389/fpubh.2019.00418},
pmid = {32039129},
issn = {2296-2565},
abstract = {Background: Particulate matter (PM) air pollution causes deleterious health effects; however, less is known about health effects of indoor air particulate matter (IAP). Objective: To understand whether IAP influences distinct mechanisms in the development of respiratory tract infections, including bacterial growth, biofilm formation, and innate immunity. Additionally, we tested whether IAP from Iowa houses of subjects with and without recent respiratory exacerbations recapitulated the National Institute of Standards and Technology (NIST) IAP findings. Methods: To test the effect of NIST and Iowa IAP on bacterial growth and biofilm formation, we assessed Staphylococcus aureus growth and Pseudomonas aeruginosa biofilm formation with and without the presence of IAP. To assess the effect of IAP on innate immunity, we exposed primary human airway surface liquid (ASL) to NIST, and Iowa IAP. Lastly, we tested whether specific metals may be responsible for effects on airway innate immunity. Results: NIST and Iowa IAP significantly enhanced bacterial growth and biofilm formation. NIST IAP (whole particle and the soluble portion) impaired ASL antimicrobial activity. IAP from one Iowa home significantly impaired ASL antimicrobial activity (p < 0.05), and five other homes demonstrated a trend (p ≤ 0.18) of impaired ASL antimicrobial activity. IAP from homes of subjects with a recent history of respiratory exacerbation tended (p = 0.09) to impair ASL antimicrobial activity more than IAP from homes of those without a history respiratory exacerbation. Aluminum and Magnesium impaired ASL antimicrobial activity, while copper was bactericidal. Combining metals varied their effect on ASL antimicrobial activity. Conclusions: NIST IAP and Iowa IAP enhanced bacterial growth and biofilm formation. ASL antimicrobial activity was impaired by NIST IAP, and Iowa house IAP from subjects with recent respiratory exacerbation tended to impair ASL antimicrobial activity. Individual metals may explain impaired ASL antimicrobial activity; however, antimicrobial activity in the presence of multiple metals warrants further study.},
}

@article {pmid32038604,
year = {2020},
author = {Dahyot, S and Oxaran, V and Niepceron, M and Dupart, E and Legris, S and Destruel, L and Didi, J and Clamens, T and Lesouhaitier, O and Zerdoumi, Y and Flaman, JM and Pestel-Caron, M},
title = {Role of the LytSR Two-Component Regulatory System in Staphylococcus lugdunensis Biofilm Formation and Pathogenesis.},
journal = {Frontiers in microbiology},
volume = {11},
number = {},
pages = {39},
doi = {10.3389/fmicb.2020.00039},
pmid = {32038604},
issn = {1664-302X},
abstract = {Staphylococcus lugdunensis is a coagulase negative Staphylococcus recognized as a virulent pathogen. It is responsible for a wide variety of infections, some of which are associated with biofilm production, such as implanted medical device infections or endocarditis. However, little is known about S. lugdunensis regulation of virulence factor expression. Two-component regulatory systems (TCS) play a critical role in bacterial adaptation, survival, and virulence. Among them, LytSR is widely conserved but has variable roles in different organisms, all connected to metabolism or cell death and lysis occurring during biofilm development. Therefore, we investigated here the functions of LytSR in S. lugdunensis pathogenesis. Deletion of lytSR in S. lugdunensis DSM 4804 strain did not alter either susceptibility to Triton X-100 induced autolysis or death induced by antibiotics targeting cell wall synthesis. Interestingly, ΔlytSR biofilm was characterized by a lower biomass, a lack of tower structures, and a higher rate of dead cells compared to the wild-type strain. Virulence toward Caenorhabditis elegans using a slow-killing assay was significantly reduced for the mutant compared to the wild-type strain. By contrast, the deletion of lytSR had no effect on the cytotoxicity of S. lugdunensis toward the human keratinocyte cell line HaCaT. Transcriptional analyses conducted at mid- and late-exponential phases showed that lytSR deletion affected the expression of 286 genes. Most of them were involved in basic functions such as the metabolism of amino acids, carbohydrates, and nucleotides. Furthermore, LytSR appeared to be involved in the regulation of genes encoding known or putative virulence and colonization factors, including the fibrinogen-binding protein Fbl, the major autolysin AtlL, and the type VII secretion system. Overall, our data suggest that the LytSR TCS is implicated in S. lugdunensis pathogenesis, through its involvement in biofilm formation and potentially by the control of genes encoding putative virulence factors.},
}

@article {pmid32038515,
year = {2019},
author = {Hegde, SR},
title = {Computational Identification of the Proteins Associated With Quorum Sensing and Biofilm Formation in Mycobacterium tuberculosis.},
journal = {Frontiers in microbiology},
volume = {10},
number = {},
pages = {3011},
doi = {10.3389/fmicb.2019.03011},
pmid = {32038515},
issn = {1664-302X},
abstract = {With prolonged therapy and increased instances of drug resistance, tuberculosis is viewed as a serious infectious disease causing high mortality. Emerging concepts in Mycobacterium tuberculosis pathogenicity include biofilm formation, which endows bacterial survival in the host for a long time. To tackle chronic tuberculosis infection, a detailed understanding of the bacterial survival mechanisms is crucial. Using comparative genomics and literature mining, 115 M. tuberculosis proteins were shortlisted for their likely association with biofilm formation or quorum sensing. These include essential genes such as secA2, lpqY-sugABC, Rv1176c, and Rv0195, many of which are also known virulence factors. Furthermore, the functional relationship among these proteins was established by considering known protein-protein interactions, regulatory interactions, and gene expression correlation data/information. Graph centrality and motif analyses predicted the importance of proteins, such as Rv0081, DevR, RegX3, Rv0097, and Rv1996 in M. tuberculosis biofilm formation. Analysis of conservation across other biofilm-forming bacteria suggests that most of these genes are conserved in mycobacteria. As the processes, such as quorum sensing, leading to biofilm formation involve diverse pathways and interactions between proteins, these system-wide studies provide a novel perspective toward understanding mycobacterial persistence.},
}

@article {pmid32037796,
year = {2020},
author = {Liu, T and Khai Lim, Z and Chen, H and Hu, S and Yuan, Z and Guo, J},
title = {Temperature-tolerated mainstream nitrogen removal by anammox and nitrite/nitrate-dependent anaerobic methane oxidation in a membrane biofilm reactor.},
journal = {Environmental science & technology},
volume = {},
number = {},
pages = {},
doi = {10.1021/acs.est.9b05650},
pmid = {32037796},
issn = {1520-5851},
abstract = {The mainstream anaerobic ammonium oxidation (anammox) process provides strong support to the on-going paradigm shift from energy-negative to energy-neutral in wastewater treatment plants. However, the low temperature (e.g., below 15oC) represents one of the major challenges for mainstream anammox in practice. In this study, a stable nitrogen removal rate (0.13 kg m-3 d-1), together with a high-level effluent quality (<5.0 mg N L-1) were achieved in a lab-scale upflow membrane biofilm reactor (MBfR) by coupling anammox with nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) microorganisms, at temperature as low as 10oC. With temperature progressively decreased from 25 to 10oC, the total nitrogen removal efficiency maintained in the range of 90-94% at a constant hydraulic retention time of 9 h. The temperature impacts on the biofilm system coupling anammox and n-DAMO reactions were increasing at a lower temperature range with higher Arrhenius coefficients. Additionally, 16S rRNA gene sequencing results showed that anammox bacteria, n-DAMO bacteria and n-DAMO archaea jointly dominated the biofilm, and their respective abundances remained relatively stable when the temperature was decreased. The major reason for this temperature-tolerated performance is the overcapacity developed, which is indicated by biofilm thickness measurements and mathematical modelling. The stable performance obtained in this study shows promise for the n-DAMO application in domestic wastewater.},
}

@article {pmid32037588,
year = {2020},
author = {Wei, D and Zhao, XH},
title = {Calcium maintained higher quality and enhanced resistance against chilling stress by regulating enzymes in reactive oxygen and biofilm metabolism of Chinese winter jujube fruit.},
journal = {Journal of food biochemistry},
volume = {},
number = {},
pages = {e13161},
doi = {10.1111/jfbc.13161},
pmid = {32037588},
issn = {1745-4514},
support = {2019XY009//Key discipline foundation of Food science and engineering in Jilin agricultural science and technology institute/ ; 201860019//University Innovation and Entrepreneurship Foundation of China/ ; },
abstract = {The postharvest senescence of Chinese winter jujube fruit can be effectively delayed by refrigerated storage. However, chilling injury often occurs in jujube fruit during cold storage. In this study, Chinese winter jujubes were sprayed with CaCl2 (4%) 3 times at intervals of 2 hr on the day of refrigeration. The results presented that maximum difference of 2.7 N firmness, 3.42% TAC, and 0.8 OD280 /g polyphenol content were detected in calcium-treated fruit during cold storage, but the levels of O 2 - , MDA, hydrogen peroxide, browning rate, electrolyte leakage, and weight loss rate were significantly inhibited (p < .05). The maximum difference of enzymes activity of CAT, POD, SOD was 2.1, 10.8, and 40.6 mol h-1 kg-1 respectively, but 21.1 mol h-1 kg-1 PPO was restrained in the treated group. In conclusion, the results provided a reliable method for inhibiting cold injury and explained the internal molecular mechanism of the fruit regulated by calcium. PRACTICAL APPLICATIONS: Refrigerated storage is an important method for extending the storage time of Chinese winter jujube fruit. However, cold damage may occur when the jujubes are stored at low temperature for long-term. It is, therefore, of great significance to find a new method and reveal the molecular mechanism. We believe that our study makes a significant contribution to the literature because it provides an effective method of maintaining higher quality and mechanistic insights into the resistance against the chilling stress of jujubes.},
}

@article {pmid32036742,
year = {2020},
author = {Skariyachan, S and Gopal, D and Kadam, SP and Muddebihalkar, AG and Uttarkar, A and Niranjan, V},
title = {Carbon fullerene act as potential lead molecule against prospective molecular targets of biofilm-producing multi-drug resistant Acinetobacter baumanni and Pseudomonas aerugenosa: Computational modeling and MD simulation studies.},
journal = {Journal of biomolecular structure & dynamics},
volume = {},
number = {},
pages = {1-36},
doi = {10.1080/07391102.2020.1726821},
pmid = {32036742},
issn = {1538-0254},
abstract = {This study aimed to screen putative drug targets associated with biofilm formation of multidrug-resistant Acinetobacter baumannii and Pseudomonas areugenosa and prioritize carbon nano-fullerene as potential lead molecule by structure-based virtual screening. Based on the functional role, 36 and 83 genes that are involved in biofilm formation of A. baumannii and P. areugenosa respectively were selected and metabolic network was computationally constructed. The genes that lack three-dimensional structures were predicted and validated. Carbon nano-fullerene selected as lead molecule and their drug-likeliness and pharmacokinetics properties were computationally predicted. The binding potential of carbon nano-fullerene towards selected drug targets was modeled and compared with the binding of conventional drugs, doripenem, and polymyxin-B with their usual targets. The stabilities of four best-docked complexes were confirmed by molecular dynamic (MD) simulation. This study suggested that selected genes demonstrated relevant interactions in the constructed metabolic pathways. Carbon fullerene exhibited significant binding abilities to most of the prioritized targets in comparison with the binding of last-resort antibiotics and their usual target. The four best ligand-receptor interactions predicted by molecular docking revealed that stability throughout MD simulation. Notably, carbon fullerene exhibited profound binding with outer membrane protein (OmpA) and ribonuclease-HII (rnhB) of A. baumannii and 2-heptyl-4(1H)-quinolone synthase (pqsBC) and chemotaxis protein (wspA) of P. aeruginosa. Thus, the current study suggested that carbon fullerene probably used as potential lead molecules towards selected targets of A. baumannii and P. aeruginosa and the applied aspects probably scaled up to design promising lead molecules towards these pathogens.},
}

@article {pmid32036275,
year = {2020},
author = {Brescia, F and Marchetti-Deschmann, M and Musetti, R and Perazzolli, M and Pertot, I and Puopolo, G},
title = {The rhizosphere signature on the cell motility, biofilm formation and secondary metabolite production of a plant-associated Lysobacter strain.},
journal = {Microbiological research},
volume = {234},
number = {},
pages = {126424},
doi = {10.1016/j.micres.2020.126424},
pmid = {32036275},
issn = {1618-0623},
abstract = {Lysobacter spp. are common bacterial inhabitants of the rhizosphere of diverse plant species. However, the impact of the rhizosphere conditions on their physiology is still relatively understudied. To provide clues on the behaviour of Lysobacter spp. in this ecological niche, we investigated the physiology of L. capsici AZ78 (AZ78), a biocontrol strain isolated from tobacco rhizosphere, on a common synthetic growth medium (LBA) and on a growth medium containing components of the plant rhizosphere (RMA). The presence of a halo surrounding the AZ78 colony on RMA was a first visible effect related to differences in growth medium composition and it corresponded to the formation of a large outer ring. The lower quantity of nutrients available in RMA as compared with LBA was associated to a higher expression of a gene encoding cAMP-receptor-like protein (Clp), responsible for cell motility and biofilm formation regulation. AZ78 cells on RMA were motile, equipped with cell surface appendages and organised in small groups embedded in a dense layer of fibrils. Metabolic profiling by mass spectrometry imaging revealed increased diversity of analytes produced by AZ78 on RMA as compared with LBA. In particular, putative cyclic lipodepsipeptides, polycyclic tetramate macrolactams, cyclic macrolactams and other putative secondary metabolites with antibiotic activity were identified. Overall, the results obtained in this study shed a light on AZ78 potential to thrive in the rhizosphere by its ability to move, form biofilm and release secondary metabolites.},
}

@article {pmid32034730,
year = {2020},
author = {Benmouna, Z and Dalache, F and Zadi-Karam, H and Karam, NE and Vuotto, C},
title = {Ability of Three Lactic Acid Bacteria to Grow in Sessile Mode and to Inhibit Biofilm Formation of Pathogenic Bacteria.},
journal = {Advances in experimental medicine and biology},
volume = {},
number = {},
pages = {},
doi = {10.1007/5584_2020_495},
pmid = {32034730},
issn = {0065-2598},
abstract = {In this study, we explored the effect of three lactic acid bacteria (LAB), i.e. Enterococcus sp CM9, Enterococcus sp CM18 and Enterococcus faecium H3, and their supernatants, on seven biofilm-forming pathogenic strains isolated from human urinary tract or nose infections. By quantitative biofilm production assay, a strong adherence ability of Enterococcus sp CM9 and Enterococcus sp CM18 was revealed while E. faecium H3 resulted to be moderately adherent. Inhibition tests demonstrated an antimicrobial activity of LAB against pathogens.The presence of cell free supernatant (CFS) of CM9 and CM18 strains significantly decreased the adhesion of S. aureus 10,850, S. epidermidis 4,296 and E. coli FSL24. The CFS of H3 strain was effective against S. epidermidis 4,296 and P. aeruginosa PA1FSL biofilms only. Biofilm formation of K. pneumoniae Kp20FSL, A. baumannii AB8FSL and ESBL+ E. coli FS101570 have not been affected by any CSF while P. aeruginosa PA1FSL biofilm increase in presence of CM9 and CM18 CFS.Confocal Laser Scanning Microscopy revealed that K. pneumoniae Kp20FSL biofilm was inhibited by Enterococcus sp CM9, when grown together.Our results suggest that the LAB strains and/or their bacteriocins can be considered as potential tools to control biofilm formation of some bacterial pathogens.},
}

@article {pmid32034427,
year = {2020},
author = {Röhner, E and Jacob, B and Böhle, S and Rohe, S and Löffler, B and Matziolis, G and Zippelius, T},
title = {Sodium hypochlorite is more effective than chlorhexidine for eradication of bacterial biofilm of staphylococci and Pseudomonas aeruginosa.},
journal = {Knee surgery, sports traumatology, arthroscopy : official journal of the ESSKA},
volume = {},
number = {},
pages = {},
doi = {10.1007/s00167-020-05887-9},
pmid = {32034427},
issn = {1433-7347},
support = {S 01/17//Stiftung Endoprothetik/ ; },
abstract = {PURPOSE: Periprosthetic infection is a common reason for surgical revision. Given the increasing resistance of bacteria to antibiotics (e.g., VRE, 4-MRGN) local antiseptic treatment is gaining in importance. However, no standard guideline-based treatment recommendation is yet available. The aim of this study was to investigate the effectiveness of sodium hypochlorite and chlorhexidine against bacterial biofilms. Furthermore, the toxicity of both antiseptics towards human chondrocytes was examined.

METHODS: Human chondrocytes were isolated, cultivated and treated with sodium hypochlorite and chlorhexidine. The viability of cultures was assessed by determination of cell count, XTT and MTT ELISAs, and fluorescent staining with propidium iodide. Bacterial strains of Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa were added to liquid media and incubated overnight. After determination of bacterial concentrations polyethylene (PE) devices were inoculated with bacteria for 48 h until biofilms formed. The devices were then washed, treated with antiseptics for 2 and 5 min and subsequently spread on agar plates.

RESULTS: Sodium hypochlorite is more effective than chlorhexidine in penetrating biofilms of S. aureus, S. epidermidis and P. aeruginosa. Both antiseptics are chondrotoxic, but sodium hypochlorite damages human chondrocytes less than chlorhexidine in vitro.

CONCLUSIONS: The findings confirm the effectiveness of sodium hypochlorite and chlorhexidine against bacterial biofilms. Both antiseptics can be recommended for the treatment of periprosthetic infections. The toxic effects of sodium hypochlorite and chlorhexidine towards chondrocytes may mean there is a risk of damage to cartilage tissue.

LEVEL OF EVIDENCE: Controlled experimental study.},
}

@article {pmid32033878,
year = {2020},
author = {Lawrence, JA and Huang, Z and Rathinavelu, S and Hu, JF and Garo, E and Ellis, M and Norman, VL and Buckle, R and Williams, RB and Starks, CM and Eldridge, GR},
title = {Optimized plant compound with potent anti-biofilm activity across gram-negative species.},
journal = {Bioorganic & medicinal chemistry},
volume = {},
number = {},
pages = {115229},
doi = {10.1016/j.bmc.2019.115229},
pmid = {32033878},
issn = {1464-3391},
abstract = {Many human diseases, including cystic fibrosis lung infections, are caused or exacerbated by bacterial biofilms. Specialized modes of motility, including swarming and twitching, allow gram-negative bacteria to spread across surfaces and form biofilms. Compounds that inhibit these motilities could slow the spread of biofilms, thereby allowing antibiotics to work better. We previously demonstrated that a set of plant-derived triterpenes, including oleanolic acid and ursolic acid, inhibit formation of Escherichia coli and Pseudomonas aeruginosa biofilms, and alter expression of genes involved in chemotaxis and motility. In the present study, we have prepared a series of analogs of oleanolic acid. The analogs were evaluated against clinical isolates of E. coli and P. aeruginosa in biofilm formation assays and swarming assays. From these analogs, compound 9 was selected as a lead compound for further development. Compound 9 inhibits E. coli biofilm formation at 4 µg/mL; it also inhibits swarming at ≤1 µg/mL across multiple clinical isolates of P. aeruginosa, E. coli, Burkholderia cepacia, and Salmonella enterica, and at <0.5 µg/mL against multiple agricultural strains. Compound 9 also potentiates the activity of the antibiotics tobramycin and colistin against swarming P. aeruginosa; this is notable, as tobramycin and colistin are inhaled antibiotics commonly used to treat P. aeruginosa lung infections in people with cystic fibrosis. qPCR experiments suggested that 9 alters expression of genes involved in regulating Type IV pili; western blots confirmed that expression of Type IV pili components PilA and PilY1 decreases in P. aeruginosa in the presence of 9.},
}

@article {pmid32033530,
year = {2020},
author = {Zheng, J and Wu, Y and Lin, Z and Wang, G and Jiang, S and Sun, X and Tu, H and Yu, Z and Qu, D},
title = {ClpP participates in stress tolerance, biofilm formation, antimicrobial tolerance, and virulence of Enterococcus faecalis.},
journal = {BMC microbiology},
volume = {20},
number = {1},
pages = {30},
doi = {10.1186/s12866-020-1719-9},
pmid = {32033530},
issn = {1471-2180},
support = {SMGC201705029//Sanming Project of Medicine in Shenzhen/ ; JCYJ20170412143551332//Shenzhen Science and Technology Innovation Commission/ ; JCYJ20180302144431923//Shenzhen Science and Technology Innovation Commission/ ; JCYJ20180508162403996//Shenzhen Science and Technology Innovation Commission/ ; JCYJ20180302144721183//Shenzhen Science and Technology Innovation Commission/ ; 2019027//Shenzhen Nanshan District Scientific Research Program of the People's Republic of China/ ; },
abstract = {BACKGROUND: ClpP is important for bacterial growth and plays an indispensable role in cellular protein quality control systems by refolding or degrading damaged proteins, but the physiological significance of ClpP in Enterococcus faecalis remains obscure. A clpP deletion mutant (△clpP) was constructed using the E. faecalis OG1RF strain to clarify the effect of ClpP on E. faecalis. The global abundance of proteins was determined by a mass spectrometer with tandem mass tag labeling.

RESULTS: The ΔclpP mutant strain showed impaired growth at 20 °C or 45 °C at 5% NaCl or 2 mM H2O2. The number of surviving ΔclpP mutants decreased after exposure to the high concentration (50× minimal inhibitory concentration) of linezolid or minocycline for 96 h. The ΔclpP mutant strain also demonstrated decreased biofilm formation but increased virulence in a Galleria mellonella model. The mass spectrometry proteomics data indicated that the abundances of 135 proteins changed (111 increased, 24 decreased) in the ΔclpP mutant strain. Among those, the abundances of stress response or virulence relating proteins: FsrA response regulator, gelatinase GelE, regulatory protein Spx (spxA), heat-inducible transcription repressor HrcA, transcriptional regulator CtsR, ATPase/chaperone ClpC, acetyl esterase/lipase, and chaperonin GroEL increased in the ΔclpP mutant strain; however, the abundances of ribosomal protein L4/L1 family protein (rplD), ribosomal protein L7/L12 (rplL2), 50S ribosomal protein L13 (rplM), L18 (rplR), L20 (rplT), 30S ribosomal protein S14 (rpsN2) and S18 (rpsR) all decreased. The abundances of biofilm formation-related adapter protein MecA increased, while the abundances of dihydroorotase (pyrC), orotate phosphoribosyltransferase (pyrE), and orotidine-5'-phosphate decarboxylase (pyrF) all decreased in the ΔclpP mutant strain.

CONCLUSION: The present study demonstrates that ClpP participates in stress tolerance, biofilm formation, antimicrobial tolerance, and virulence of E. faecalis.},
}

@article {pmid32033480,
year = {2020},
author = {Leonetti, S and Tuvo, B and Campanella, B and Legnaioli, S and Onor, M and Bramanti, E and Totaro, M and Baggiani, A and Giorgi, S and Privitera, GP and Piolanti, N and Parchi, PD and Casini, B},
title = {Evaluation of Microbial Adhesion and Biofilm Formation on Nano-Structured and Nano-Coated Ortho-Prosthetic Materials by a Dynamic Model.},
journal = {International journal of environmental research and public health},
volume = {17},
number = {3},
pages = {},
doi = {10.3390/ijerph17031013},
pmid = {32033480},
issn = {1660-4601},
support = {PRA 2017/2018//This work was supported by the University of Pisa Research Project/ ; },
abstract = {The bio-engineering technologies of medical devices through nano-structuring and coating was recently proposed to improve biocompatibility and to reduce microbial adhesion in the prevention of implantable device-related infections. Our aim was to evaluate the ability of new nano-structured and coated materials to prevent the adhesion and biofilm formation, according to the American Standard Test Method ASTM-E2647-13. The materials composition was determined by X-ray Fluorescence and Laser Induced Breakdown Spectroscopy. Silver release was evaluated by Inductively Coupled Plasma Mass Spectrometry analysis. The gene expression levels of the Quorum Sensing Las and Rhl system were evaluated by the ΔΔCt method. The Log bacterial density (Log CFU/cm2) on TiAl6V4 was 4.41 ± 0.76 and 4.63 ± 1.01 on TiAl6V4-AgNPs compared to 2.57 ± 0.70 on CoCr and 2.73 ± 0.61 on CoCr-AgNPs (P < 0.0001, A.N.O.V.A.- one way test). The silver release was found to be equal to 17.8 ± 0.2 µg/L after the batch phase and 1.3 ± 0.1 µg/L during continuous flow. The rhlR gene resulted in a 2.70-fold increased expression in biofilm growth on the silver nanoparticles (AgNPs) coating. In conclusion, CoCr showed a greater ability to reduce microbial adhesion, independently of the AgNPs coating. The silver release resulted in promoting the up-regulation of the Rhl system. Further investigation should be conducted to optimize the effectiveness of the coating.},
}

@article {pmid32033098,
year = {2020},
author = {Niu, TX and Wang, XN and Wu, HY and Bi, JR and Hao, HS and Hou, HM and Zhang, GL},
title = {Transcriptomic Analysis, Motility and Biofilm Formation Characteristics of Salmonella typhimurium Exposed to Benzyl Isothiocyanate Treatment.},
journal = {International journal of molecular sciences},
volume = {21},
number = {3},
pages = {},
doi = {10.3390/ijms21031025},
pmid = {32033098},
issn = {1422-0067},
support = {No.2017YFC1600403//The National Key R and D Program of China/ ; 31571888//The National Natural Science Foundation of China/ ; 2019-MS-021//Liaoning Provincial Natural Science Foundation of China/ ; },
abstract = {Salmonella typhimurium (S. typhimurium) is a common foodborne pathogen that not only causes diseases and contaminates food, but also causes considerable economic losses. Therefore, it is necessary to find effective and feasible methods to control S. typhimurium. In this study, changes in S. typhimurium after treatment with benzyl isothiocyanate (BITC) were detected by transcriptomics to explore the antibacterial effect of BITC at subinhibitory concentration. The results showed that, in contrast to the control group (SC), the BITC-treated group (SQ_BITC) had 197 differentially expressed genes (DEGs), of which 115 were downregulated and 82 were upregulated. We screened out eight significantly downregulated virulence-related genes and verified gene expression by quantitative Real-time Polymerase Chain Reaction (qRT-PCR). We also selected motility and biofilm formation to observe the effects of BITC on the other virulence related factors of S. typhimurium. The results showed that both swimming and swarming were significantly inhibited. BITC also had a significant inhibitory effect on biofilm formation, and showed an effect on bacterial morphology. These results will be helpful for understanding the mechanism of the antibacterial action of BITC against S. typhimurium and other foodborne pathogens.},
}

@article {pmid32032989,
year = {2020},
author = {Anticó, E and Fontàs, C and Vera, R and Mostazo, G and Salvadó, V and Guasch, H},
title = {A novel Cyphos IL 104-based polymer inclusion membrane (PIM) probe to mimic biofilm zinc accumulation.},
journal = {The Science of the total environment},
volume = {715},
number = {},
pages = {136938},
doi = {10.1016/j.scitotenv.2020.136938},
pmid = {32032989},
issn = {1879-1026},
abstract = {The presence of Zn in surface waters from abandoned mining zones is a critical issue since excess Zn concentrations may affect aquatic life and whole ecosystems. We present, for the first time, a simple tool based on a polymer inclusion membrane (PIM) intended to monitor Zn in river water by mimicking metal accumulation in the biofilm. The PIM-based probe contains a polymeric membrane prepared using cellulose triacetate (CTA, 50% w/w) as the base polymer, nitrophenyloctyl ether (NPOE) as the plasticizer (20% w/w), and the ionic liquid (IL) Cyphos 104 as the extractant (30% w/w). The accumulation of Zn in the acceptor phase (0.01 M HNO3) was evaluated for different free metal concentrations at 4 h accumulation time resulting in a good correlation between the free metal concentration and the accumulated one. We also found that the metal accumulated agrees with the free metal fraction upon addition of EDTA in the donor solution. The results for Zn accumulation with the PIM-based probe were found to be comparable to those obtained for a biofilm that was grown in a stream from an abandoned mine area and subsequently translocated to the laboratory and put in contact with Zn polluted stream water, so confirming the effectiveness of this new probe in mimicking Zn accumulation in the biofilm.},
}

@article {pmid32032938,
year = {2020},
author = {Hong, P and Wu, X and Shu, Y and Wang, C and Tian, C and Wu, H and Xiao, B},
title = {Bioaugmentation treatment of nitrogen-rich wastewater with a denitrifier with biofilm-formation and nitrogen-removal capacities in a sequencing batch biofilm reactor.},
journal = {Bioresource technology},
volume = {303},
number = {},
pages = {122905},
doi = {10.1016/j.biortech.2020.122905},
pmid = {32032938},
issn = {1873-2976},
abstract = {A strain with efficient biofilm-formation and aerobic denitrification capabilities was isolated and identified as Pseudomonas mendocina IHB602. In pure culture, strain IHB602 removed almost all NO3--N, NO2--N, and NH4+-N (initial concentrations 50 mg/L) within 24 h. The strain produced large amounts of extracellular polymeric substances (maximum 430.33 mg/g cell dry weight) rich in protein but containing almost no humic acid. This, and strong autoaggregation (maximum 47.09%) and hydrophobicity (maximum 85.07%), imparted strain IHB602 with biofilm forming traits. A sequencing batch biofilm reactor bioaugmented with strain IHB602 (SBBR1) had more rapid biofilm-formation than the control without strain IHB602 inoculation (SBBR2). During the stabilization period, the effluent removal ratios for NH4+-N (95%), NO3--N (91%) and TN (88%) in SBBR1 were significantly higher than those in SBBR2 (NH4+-N: 91%, NO3--N: 88%, TN: 82%). Microbial community structure analysis revealed that strain IHB602 successfully proliferated and contributed to nitrogen removal as well as biofilm formation.},
}

@article {pmid32032775,
year = {2020},
author = {Liu, T and Guo, J and Hu, S and Yuan, Z},
title = {Model-based investigation of membrane biofilm reactors coupling anammox with nitrite/nitrate-dependent anaerobic methane oxidation.},
journal = {Environment international},
volume = {137},
number = {},
pages = {105501},
doi = {10.1016/j.envint.2020.105501},
pmid = {32032775},
issn = {1873-6750},
abstract = {An innovative process coupling anaerobic ammonium oxidation (anammox) with nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) in membrane biofilm reactors (MBfRs) has been developed to achieve high-level nitrogen removal from both sidestream (i.e., anaerobic digestion liquor) and mainstream (i.e., domestic strength) wastewater. In this study, a 1D biofilm model embedding the n-DAMO and anammox reactions was developed to facilitate further understanding of the process and its optimization. The model was calibrated and validated using comprehensive data sets from two independent MBfRs, treating sidestream- and mainstream-strength wastewater, respectively. Modelling results revealed a unique biofilm stratification. While anammox bacteria dominated throughout the biofilm, n-DAMO archaea (coupling nitrate reduction with anaerobic methane oxidation) only occurred at the inner layer and n-DAMO bacteria (coupling nitrite reduction with anaerobic methane oxidation) spread more evenly with a slightly higher fraction in the outer layer. The established MBfRs were robust against dynamic influent flowrates and nitrite/ammonium ratios. Thicker biofilms were beneficial for not only the total nitrogen (TN) removal but also the system robustness. Additionally, a positive correlation between the nitrogen removal efficiency and the residual methane emission was observed, as a result of higher methane partial pressure required. However, there was a threshold of methane partial pressure, above which the residual methane increased but nitrogen removal efficiency was stable. Meanwhile, thicker biofilms were also favorable to achieve less residual methane emission. Simulation results also suggested the feasibility of methane-based MBfRs to polish mainstream anammox effluent to meet a stringent N discharge standard (e.g., TN < 5 mg/L).},
}

@article {pmid32032766,
year = {2020},
author = {Chen, L and Gu, L and Geng, X and Xu, G and Huang, X and Zhu, X},
title = {A novel cis antisense RNA AsfD promotes Salmonella enterica serovar Typhi motility and biofilm formation.},
journal = {Microbial pathogenesis},
volume = {},
number = {},
pages = {104044},
doi = {10.1016/j.micpath.2020.104044},
pmid = {32032766},
issn = {1096-1208},
abstract = {Bacterial non-coding RNAs (ncRNAs) can participate in multiple biological processes, including motility, biofilm formation, and virulence. Using high-throughput sequencing and transcriptome analysis of Salmonella enterica serovar Typhi (S. Typhi), we identified a novel antisense RNA located at the opposite strand of the flhDC operon. In this study, a northern blot and qRT-PCR were used to confirm the expression of this newfound antisense RNA in S. Typhi. Moreover, 5' RACE and 3' RT-PCR were performed to reveal the molecular characteristics of the antisense RNA, which was 2079 nt - 2179 nt in length, covered the entire flhDC operon sequence, and termed AsfD. The level of AsfD expression was higher during the stationary phase of S. Typhi and activated by the regulators, OmpR and Fis. When AsfD was overexpressed, the level of flagellar gene flhDC transcription increased; moreover, the level of fliA and fljB expression, as well as the motility and biofilm formation of S. Typhi were also enhanced. The results of this study suggest that AsfD is likely to enhance the motility and biofilm formation of S. Typhi by up-regulating flhDC expression.},
}

@article {pmid32029764,
year = {2020},
author = {Ahmad, I and Nygren, E and Khalid, F and Myint, SL and Uhlin, BE},
title = {A Cyclic-di-GMP signalling network regulates biofilm formation and surface associated motility of Acinetobacter baumannii 17978.},
journal = {Scientific reports},
volume = {10},
number = {1},
pages = {1991},
doi = {10.1038/s41598-020-58522-5},
pmid = {32029764},
issn = {2045-2322},
support = {8666/Punjab/NRPU/R&D/HEC/2017//Higher Education Commission, Pakistan (HEC)/ ; 2015-03007//Vetenskapsrådet (Swedish Research Council)/ ; 2015-06824//Vetenskapsrådet (Swedish Research Council)/ ; 2016-06598//Vetenskapsrådet (Swedish Research Council)/ ; 349-2007-8673//Vetenskapsrådet (Swedish Research Council)/ ; 829-2006-7431//Vetenskapsrådet (Swedish Research Council)/ ; JCK-1527//Kempestiftelserna (Kempe Foundations)/ ; JCK-1724//Kempestiftelserna (Kempe Foundations)/ ; },
abstract = {Acinetobacter baumannii has emerged as an increasing multidrug-resistant threat in hospitals and a common opportunistic nosocomial pathogen worldwide. However, molecular details of the pathogenesis and physiology of this bacterium largely remain to be elucidated. Here we identify and characterize the c-di-GMP signalling network and assess its role in biofilm formation and surface associated motility. Bioinformatic analysis revealed eleven candidate genes for c-di-GMP metabolizing proteins (GGDEF/EAL domain proteins) in the genome of A. baumannii strain 17978. Enzymatic activity of the encoded proteins was assessed by molecular cloning and expression in the model organisms Salmonella typhimurium and Vibrio cholerae. Ten of the eleven GGDEF/EAL proteins altered the rdar morphotype of S. typhimurium and the rugose morphotype of V. cholerae. The over expression of three GGDEF proteins exerted a pronounced effect on colony formation of A. baumannii on Congo Red agar plates. Distinct panels of GGDEF/EAL proteins were found to alter biofilm formation and surface associated motility of A. baumannii upon over expression. The GGDEF protein A1S_3296 appeared as a major diguanylate cyclase regulating macro-colony formation, biofilm formation and the surface associated motility. AIS_3296 promotes Csu pili mediated biofilm formation. We conclude that a functional c-di-GMP signalling network in A. baumannii regulates biofilm formation and surface associated motility of this increasingly important opportunistic bacterial pathogen.},
}

@article {pmid32029738,
year = {2019},
author = {Vermilyea, DM and Ottenberg, GK and Davey, ME},
title = {Citrullination mediated by PPAD constrains biofilm formation in P. gingivalis strain 381.},
journal = {NPJ biofilms and microbiomes},
volume = {5},
number = {1},
pages = {7},
doi = {10.1038/s41522-019-0081-x},
pmid = {32029738},
issn = {2055-5008},
support = {R01DEO24580//U.S. Department of Health & Human Services | NIH | National Institute of Dental and Craniofacial Research (NIDCR)/ ; R01DE019117//U.S. Department of Health & Human Services | NIH | National Institute of Dental and Craniofacial Research (NIDCR)/ ; F31DE027278//U.S. Department of Health & Human Services | NIH | National Institute of Dental and Craniofacial Research (NIDCR)/ ; R01DEO24580//U.S. Department of Health & Human Services | NIH | National Institute of Dental and Craniofacial Research (NIDCR)/ ; R01DEO19117//U.S. Department of Health & Human Services | NIH | National Institute of Dental and Craniofacial Research (NIDCR)/ ; },
abstract = {Porphyromonas gingivalis is the only known human-associated prokaryote that produces a peptidylarginine deiminase (PPAD), a protein-modifying enzyme that is secreted along with a number of virulence factors via a type IX secretion system (T9SS). While the function of PPAD in P. gingivalis physiology is not clear, human peptidylarginine deiminases are known to convert positively charged arginine residues within proteins to neutral citrulline and, thereby, impact protein conformation and function. Here, we report that the lack of citrullination in a PPAD deletion mutant (Δ8820) enhances biofilm formation. More Δ8820 cells attached to the surface than the parent strain during the early stages of biofilm development and, ultimately, mature Δ8820 biofilms were comprised of significantly more cell-cell aggregates and extracellular matrix. Imaging by electron microscopy discovered that Δ8820 biofilm cells secrete copious amounts of protein aggregates. Furthermore, gingipain-derived adhesin proteins, which are also secreted by the T9SS were predicted by mass spectrometry to be citrullinated and citrullination of these targets by wild-type strain 381 in vitro was confirmed. Lastly, Δ8820 biofilms contained more gingipain-derived adhesin proteins and more gingipain activity than 381 biofilms. Overall, our findings support the model that citrullination of T9SS cargo proteins known to play a key role in colonization, such as gingipain-derived adhesin proteins, is an underlying mechanism that modulates P. gingivalis biofilm development.},
}

@article {pmid32029309,
year = {2019},
author = {Burneo, BS and Juárez, AS and Nieto-Monteros, DA},
title = {Un-steady state modeling for free cyanide removal and biofilm growth in a RBC batch process.},
journal = {Journal of hazardous materials},
volume = {},
number = {},
pages = {120647},
doi = {10.1016/j.jhazmat.2019.05.040},
pmid = {32029309},
issn = {1873-3336},
abstract = {Biofilm growth and free cyanide biological removal from gold mine wastewater were modeled and simulated using a bench-scale rotating biological contactor (RBC). Eight batch cultures were run in three independent compartments (1.7 L, each) of the RBC. The system worked under the following conditions: [CNi-] = 0.3 g/L, pH = 10.5 ± 0.5, T = 20 ± 5 °C, ω =5 rpm, and 40.5 % of disc submersion. During each culture, biofilm thickness, biomass, and free cyanide concentration in the liquid were quantified. Subsequently, μmax, [Formula: see text] , [Formula: see text] were determined using experimental data to later model and simulate the biofilm thickness and free cyanide biological removal with Wolfram Mathematica software. After the experiments, free cyanide biological removal was 96.33 % after three days, and maximum biofilm thickness was 0.0292 cm in the 16th day. Moreover, biofilm growth and free cyanide consumption models were adjusted to the experimental data with r2 = 0.90 and r2 = 0.99. Also, there was an equivalent error of 7.89 and 7.38 and a standard deviation of 10.89 % and 10.17 %, between the models and their experimental data, respectively. Finally, the proposed models will allow improvement of reactor operation and its design.},
}

@article {pmid32028684,
year = {2020},
author = {Vestby, LK and Grønseth, T and Simm, R and Nesse, LL},
title = {Bacterial Biofilm and its Role in the Pathogenesis of Disease.},
journal = {Antibiotics (Basel, Switzerland)},
volume = {9},
number = {2},
pages = {},
doi = {10.3390/antibiotics9020059},
pmid = {32028684},
issn = {2079-6382},
abstract = {Recognition of the fact that bacterial biofilm may play a role in the pathogenesis of disease has led to an increased focus on identifying diseases that may be biofilm-related. Biofilm infections are typically chronic in nature, as biofilm-residing bacteria can be resilient to both the immune system, antibiotics, and other treatments. This is a comprehensive review describing biofilm diseases in the auditory, the cardiovascular, the digestive, the integumentary, the reproductive, the respiratory, and the urinary system. In most cases reviewed, the biofilms were identified through various imaging technics, in addition to other study approaches. The current knowledge on how biofilm may contribute to the pathogenesis of disease indicates a number of different mechanisms. This spans from biofilm being a mere reservoir of pathogenic bacteria, to playing a more active role, e.g., by contributing to inflammation. Observations also indicate that biofilm does not exclusively occur extracellularly, but may also be formed inside living cells. Furthermore, the presence of biofilm may contribute to development of cancer. In conclusion, this review shows that biofilm is part of many, probably most chronic infections. This is important knowledge for development of effective treatment strategies for such infections.},
}

@article {pmid32028622,
year = {2020},
author = {Nett, JE and Andes, DR},
title = {Contributions of the Biofilm Matrix to Candida Pathogenesis.},
journal = {Journal of fungi (Basel, Switzerland)},
volume = {6},
number = {1},
pages = {},
doi = {10.3390/jof6010021},
pmid = {32028622},
issn = {2309-608X},
support = {112580130/DDCF/Doris Duke Charitable Foundation/United States ; R01AI073289/NH/NIH HHS/United States ; K08 AI108727/NH/NIH HHS/United States ; },
abstract = {In healthcare settings, Candida spp. cause invasive disease with high mortality. The overwhelming majority of cases are associated with the use of critically-needed medical devices, such as vascular catheters. On the surface of these indwelling materials, Candida forms resilient, adherent biofilm communities. A hallmark characteristic of this process is the production of an extracellular matrix, which promotes fungal adhesion and provides protection from external threats. In this review, we highlight the medical relevance of device-associated Candida biofilms and draw attention to the process of Candida-biofilm-matrix production. We provide an update on the current understanding of how biofilm extracellular matrix contributes to pathogenicity, particularly through its roles in the promoting antifungal drug tolerance and immune evasion.},
}

@article {pmid32027606,
year = {2018},
author = {Triveni, AG and Suresh Kumar, M and Manjunath, C and Shivannavar, CT and Gaddad, SM},
title = {Biofilm formation by clinically isolated Staphylococcus Aureus from India.},
journal = {Journal of infection in developing countries},
volume = {12},
number = {12},
pages = {1062-1066},
doi = {10.3855/jidc.10671},
pmid = {32027606},
issn = {1972-2680},
abstract = {INTRODUCTION: Staphylococcal biofilms are prominent cause for acute and chronic infection both in hospital and community settings across the world. Current study explores biofilm formation by Staphylococcus aureus isolates from clinical samples by different methods.

METHODOLOGY: Standard techniques used for the characterization of S.aureus. Qualitative and quantitative biofilm formation was assessed by Congo red Agar, Tube and Microtiter plate methods.

RESULTS: A total of 188 clinical isolates of S.aureus were screened for biofilm formation and 72 (38.29%) of them were found to be biofilm producers, 34 (18.08%) strong, 38 (20.21%) moderate. The remaining 116 (61.7%) were weak/ non biofilm producers. Maximum biofilm formers were recorded in pus samples (39.06%), followed by isolates from blood (38.23%) and urine (34.61%). Statistical analysis for the formation of biofilm indicated that Microtiter plate method is the most sensitive and specific method for screening biofilm production.

CONCLUSIONS: Biofilm formation is one of the influential virulence factor in staphylococcal pathogenesis and persistence. Microtiter plate and Congo red agar remain as reliable methods for the qualitative and quantitative estimation of biofilm formation. Monitoring of biofilm formation in various etiological agents will help in determining the severity of infection.},
}

@article {pmid32026679,
year = {2020},
author = {Barlow, DE and Biffinger, JC and Estrella, L and Lu, Q and Hung, CS and Nadeau, LJ and Crouch, AL and Russell, JN and Crookes-Goodson, WJ},
title = {Edge-Localized Biodeterioration and Secondary Microplastic Formation by Papiliotrema laurentii Unsaturated Biofilm Cells on Polyurethane Films.},
journal = {Langmuir : the ACS journal of surfaces and colloids},
volume = {},
number = {},
pages = {},
doi = {10.1021/acs.langmuir.9b03421},
pmid = {32026679},
issn = {1520-5827},
abstract = {Painted environmental surfaces are prone to microbiological colonization with potential coating deterioration induced by the microorganisms. Accurate mechanistic models of these interactions require an understanding of the heterogeneity in which the deterioration processes proceed. Here, unsaturated biofilms (i.e., at air/solid interfaces) of the yeast Papiliotrema laurentii were prepared on polyether polyurethane (PEUR) and polyester-polyether polyurethane (PEST-PEUR) coatings and incubated for up to 33 days at controlled temperature and humidity with no additional nutrients. Transmission micro-Fourier transform infrared microscopy (μFTIR) confirmed preferential hydrolysis of the ester component by the biofilm. Atomic force microscopy combined with infrared nanospectroscopy (AFM-IR) was used to analyze initial PEST-PEUR coating deterioration processes at the single-cell level, including underlying surfaces that became exposed following cell translocation. The results revealed distinct deterioration features that remained localized within ∼10 μm or less of the edges of individual cells and cell clusters. These features comprised depressions of up to ∼300 nm with locally reduced ester/urethane ratios. They are consistent with a formation process initiated by enzymatic ester hydrolysis followed by erosion from water condensation cycles. Further observations included particle accumulation in the broader biofilm vicinity. AFM-IR spectroscopy indicated these to be secondary microplastics consisting of urethane-rich oligomeric aggregates. Overall, multiple contributing factors have been identified that can facilitate differential deterioration rates across the PEST-PEUR surface. Effects of the imposed nutrient conditions on Papiliotrema laurentii physiology were also apparent, with cells developing the characteristics of starvation response, despite the availability of polyester metabolites as a carbon source. The combined results provide new laboratory insights into field-relevant microbiological polymer deterioration mechanisms and biofilm physiology at polymer coating interfaces.},
}

@article {pmid32023892,
year = {2020},
author = {de Oliveira, RVD and Bonafé, FSS and Spolidorio, DMP and Koga-Ito, CY and Farias, AL and Kirker, KR and James, GA and Brighenti, FL},
title = {Streptococcus mutans and Actinomyces naeslundii Interaction in Dual-Species Biofilm.},
journal = {Microorganisms},
volume = {8},
number = {2},
pages = {},
doi = {10.3390/microorganisms8020194},
pmid = {32023892},
issn = {2076-2607},
support = {2012/17236-4, 2013/12326-8, 2014/16935-1, 2014/02397-8 and 2015/07679-4//São Paulo Research Foundation (FAPESP)/ ; Finance Code 001//Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES)/ ; },
abstract = {The study of bacterial interaction between Streptococcus mutans and Actinomyces naeslundii may disclose important features of biofilm interspecies relationships. The aim of this study was to characterize-with an emphasis on biofilm formation and composition and metabolic activity-single- and dual-species biofilms of S. mutans or A. naeslundii, and to use a drip flow reactor (DFR) to evaluate biofilm stress responses to 0.2% chlorhexidine diacetate (CHX). Single- and dual-species biofilms were grown for 24 h. The following factors were evaluated: cell viability, biomass and total proteins in the extracellular matrix, 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide-"XTT"-reduction and lactic acid production. To evaluate stress response, biofilms were grown in DFR. Biofilms were treated with CHX or 0.9% sodium chloride (NaCl; control). Biofilms were plated for viability assessment. Confocal laser-scanning microscopy (CLSM) was also performed. Data analysis was carried out at 5% significance level. S. mutans viability and lactic acid production in dual-species biofilms were significantly reduced. S. mutans showed a higher resistance to CHX in dual-species biofilms. Total protein content, biomass and XTT reduction showed no significant differences between single- and dual-species biofilms. CLSM images showed the formation of large clusters in dual-species biofilms. In conclusion, dual-species biofilms reduced S. mutans viability and lactic acid production and increased S. mutans' resistance to chlorhexidine.},
}

@article {pmid32023715,
year = {2020},
author = {Guo, HN and Chen, Z and Xiang, J},
title = {[Influence of abaR gene knockout on growth metabolism and biofilm formation of Acinetobacter baumannii].},
journal = {Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns},
volume = {36},
number = {1},
pages = {32-36},
doi = {10.3760/cma.j.issn.1009-2587.2020.01.006},
pmid = {32023715},
issn = {1009-2587},
support = {16ZR1420800//Natural Science Foundation of Shanghai Municipal Science and Technology Commission/ ; },
abstract = {Objective: To investigate the influence of abaR gene knockout on growth metabolism and biofilm formation of Acinetobacter baumannii. Methods: The abaR gene was knocked out from Acinetobacter baumannii standard strain ATCC 17978 (wild strain) by homologous recombination method, and then the ATCC 17978 abaR knockout strain (ATCC 17978/ΔabaR: : Kn) was obtained and verified by polymerase chain reaction (PCR) electrophoresis and sequencing. The growth curves of Acinetobacter baumannii wild strain and Acinetobacter baumannii knockout strain were determined by microplate reader within cultivation hour (CH) 18, and the biofilm formation ability was measured by crystal violet staining at CH 8, 24, and 48, respectively. The sample number at each time point was 3.The results were denoted as absorbance value. Data were processed with analysis of variance of factorial design, one-way analysis of variance, t test, and least-significant difference test. Results: (1) The length of PCR product of target fragment ΔabaR: : Kn was 3 029 bp. The abaR gene was knocked out to obtain the knockout strain ATCC 17978/ΔabaR: : Kn. The length of PCR product of the knockout strain was 3 300 bp. The abaR gene was successfully knocked out. (2) At CH 2, 3, and 4, the absorbance values of Acinetobacter baumannii wild strain were slightly higher than those of the knockout strain. The absorbance values of Acinetobacter baumannii wild strain and knockout strain were similar from CH 5 to 18. (3) At CH 8 and 24, the biofilm formation ability of Acinetobacter baumannii wild strains (0.644±0.066, 0.574±0.184) was similar to that of knockout strains (0.559±0.008, 0.394±0.030, t=2.209, 1.167, P>0.05). At CH 48, the biofilm formation ability of Acinetobacter baumannii wild strains (1.157±0.259) was significantly stronger than that of Acinetobacter baumannii knockout strains (0.576±0.026, t=3.865, P<0.05). The biofilm formation ability of Acinetobacter baumannii wild strains at CH 48 was significantly stronger than that at CH 8 and 24 (P<0.05). The biofilm formation ability of Acinetobacter baumannii knockout strains at CH 24 was significantly weaker than that at CH 8 and 48 (P<0.05). Conclusions: The abaR gene of Acinetobacter baumannii ATCC 17978 can be successfully knocked out by homologous recombination to obtain its knockout strain ATCC 17978/ΔabaR: : Kn. The abaR gene does not affect the growth and metabolism of Acinetobacter baumanniibut can weaken its biofilm formation ability.},
}

@article {pmid32023713,
year = {2020},
author = {Qi, ZY and Yang, SY and Dong, SW and Zhao, FF and Qin, JH and Xiang, J},
title = {[Biological characteristics and genomic information of a bacteriophage against pan-drug resistant Klebsiella pneumoniae in a burn patient and its effects on bacterial biofilm].},
journal = {Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns},
volume = {36},
number = {1},
pages = {14-23},
doi = {10.3760/cma.j.issn.1009-2587.2020.01.004},
pmid = {32023713},
issn = {1009-2587},
support = {16ZR1420800//Natural Science Foundation of Shanghai Science and Technology Commission/ ; },
abstract = {Objective: To isolate a bacteriophage against pan-drug resistant Klebsiella pneumoniae in a burn patient, and to study its biological characteristics, genomic information, and effects on bacterial biofilm. Methods: (1) In 2018, pan-drug resistant Klebsiella pneumoniae UA168 (hereinafter referred to as the host bacteria) solution isolated from the blood of a burn patient in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (hereinafter referred to as Ruijin Hospital) was used to isolate and purify the bacteriophage against pan-drug resistant Klebsiella pneumoniae from the sewage of Ruijin Hospital with sewage co-culture method, drip plate method, and double-agar plate method. The bacteriophage was named as phage KP168 and the plaque morphology was observed. (2) The phage KP168 solution was taken for cesium chloride density gradient centrifugation and dialysis, and then the morphology of phage KP168 was observed through transmission electron microscope after phosphotungstic acid negative staining. (3) The phage KP168 solution was taken to determine the lytic ability of the phage KP168 against 20 strains of pan-drug resistant Klebsiella pneumoniae isolated from the burned patients' blood in Ruijin Hospital by the drip plate method, and then the lysis rate was calculated. (4) The phage KP168 solution at a initial titer of 9.3×10(11) plaque-forming unit (PFU)/mL (400 μL per tube) and the host bacteria solution at a concentration of 1×10(9) colony-forming unit (CFU)/mL (4 mL per tube) were conventionally shaking cultured together for 4 hours at multiplicity of infection (MOI) of 10.000, 1.000, 0.100, 0.010, or 0.001, respectively (1 tube per MOI). The titer of phage KP168 was measured by the double-agar plate method (the measurement method was the same below) to select the optimal MOI. The experiment was repeated three times. (5) The host bacteria solution at a concentration of 1×10(9) CFU/mL (4 mL per tube) and the phage KP168 solution at an adjusted titer of 5×10(7) PFU/mL (400 μL per tube) were mixed at the MOI of 0.005. The plaques were counted 0 (immediately), 1, 2, 3, 4, 5, 15, and 30 minutes (1 tube at each time point) after mixing by the double-agar plate method (the counting method was the same below), and the percentage of adsorbed phages was calculated to screen for the optimal adsorption time. The experiment was repeated three times. (6) The host bacteria solution at a concentration of 1×10(9) CFU/mL (300 μL per tube) and the phage KP168 solution at a titer of 5×10(8) PFU/mL (60 μL per tube) were mixed at MOI of 0.005 and conventionally shaking cultured after standing for the optimal adsorption time. The phage KP168 titer was measured 0 (immediately), 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 minutes after culture, and a one-step growth curve was drawn. The experiment was repeated three times. (7) The phage KP168 solution at a titer of 2.5×10(10) PFU/mL was left to stand for 1 hour at 37, 40, 50, 60, or 70 ℃ (3 tubes at each time point, 1 mL per tube) for counting the plaques, and then the thermal stability curve was drawn. SM buffer at a pH values of 5.0, 6.0, 7.0, 7.4, 8.0, 9.0, or 10.0 were added to the phage KP168 solution at a titer of 3.0×10(10) PFU/mL, respectively. The mixed solution was left to stand for 1 hour at 37 ℃ (3 tubes of each pH, each tube containing 100 μL phage KP168 solution and 900 μL SM buffer), and then the plaques were counted, and an acid-base stability curve was drawn. (8) The phage KP168 solution was taken for DNA extraction and sequencing after dialysis as in experiment (2). The whole genome was annotated with Prokka to obtain the coding sequence of phage KP168. Nucleotide's BLAST function was used to proceed nucleic acid sequence alignment for finding a known phage with the highest similarity to the phage KP168 nucleic acid sequence, and Blastx function was used to translate the coding sequence into protein for its function prediction. The comparison with Antibiotic Resistance Genes Database and Virulence Factors Database was proceeded. (9) In a 96-well plate, at a MOI of 1.000, 0.100, 0.010 or 0.001 (3 wells per MOI), 20 μL phage KP168 solution at a initial titer of 5.8×10(10) PFU/mL was added to 200 μL host bacteria solution at a concentration of 1.5×10(8) CFU/mL (the same concentration below) for co-cultivation for 48 hours. After 200 μL host bacteria solution was left to stand for 48 hours, 20 μL phage KP168 solution at a titer of 1×10(6,) 1×10(7,) 1×10(8,) 1×10(9,) or 1×10(10) PFU/mL (3 wells per titer) was added respectively for action for 4 hours. In both experiments, 200 μL host bacteria solution added with 20 μL SM buffer (3 wells) acted as a negative control, and 220 μL LB culture medium (3 wells) acted as a blank control. Absorbance values were measured by a microplate reader, and inhibition/destruction rates of biofilm were calculated. The experiments were both repeated three times. Results: (1) The plaques of phage KP168 successfully isolated and purified were transparent and round, and its diameter was approximately 1.5 mm. (2) The phage KP168 has a regular polyhedron structure with a diameter of about 50 nm and without a tail. (3) The phage KP168 could lyse 13 of 20 strains of Klebsiella pneumoniae from burned patients, with a lysis rate of 65.0%. (4) When MOI was 1.000, the titer was the highest after co-culturing the phage KP168 with the host bacteria for 4 hours, which was the optimal MOI. (5) After the mixing of the phage KP168 with the host bacteria for 4 minutes, the percentage of the adsorbed phage reached the highest, which was the optimal adsorption time. (6) The one-step growth curve showed that during the lysis of the host bacteria by phage KP168, the incubation period was about 10 minutes, and the lysis period was about 40 minutes. (7) With the condition of 40 ℃ or pH 7.4, the number of plaques and the activity of phage KP168 reached the highest. (8) The genome of phage KP168 was a linear double-stranded DNA with a length of 40 114 bp. There were 48 possible coding sequences. It had the highest similarity to Klebsiella phage_vB_Kp1. The most similar known proteins corresponding to the translated proteins of coding sequences contained 23 hypothetical proteins and 25 proteins with known functions. No resistance genes or virulence factor genes were found. The GeneBank accession number was KT367885. (9) After 48 hours of co-cultivation of the phage KP168 and the host bacteria at each MOI, the inhibition rates of biofilm were similar, with an average of about 45%. After the phage KP168 with a titer of 1×10(9) PFU/mL acted on the biofilm formed by the host bacteria for 4 h, the destruction rate of biofilm was the highest, reaching an average of 42%. Conclusions: In this study, a bacteriophage against pan-drug resistant Klebsiella pneumoniae from a burn patient, phage KP168, is isolated from sewage, which belongs to the tailless phage. It has a wide host spectrum, short adsorption time, and short incubation period, with certain thermal and acid-base stability. Its genomic information is clear, and it does not contain resistance genes or virulence factor genes. It also has an inhibitory effect on the formation of bacterial biofilm and a destructive effect on the formed bacterial biofilm.},
}

@article {pmid32012126,
year = {2018},
author = {Soltani, S and Arshadi, M and Getso, MI and Aminharati, F and Mahmoudi, M and Pourmand, MR},
title = {Prevalence of virulence genes and their association with biofilm formation in VRE faecium isolates from Ahvaz, Iran.},
journal = {Journal of infection in developing countries},
volume = {12},
number = {11},
pages = {970-977},
doi = {10.3855/jidc.10078},
pmid = {32012126},
issn = {1972-2680},
abstract = {INTRODUCTION: Vancomycin-resistant Enterococcus faecium (VREfm) is a common cause of nosocomial infections. Biofilm formation is an important factor in recurrence of infections, facilitating transfer of genetic elements, leading to treatment failures. The aim of this study was to investigate the virulence genes in biofilm producing isolates and to determine possible association between biofilm formation and the presence of these genes; also to determine association between antibiotic susceptibility patterns of VREfm isolates and their biofilm formation ability.

METHODOLOGY: A total of 57 isolates of VREfm were recovered from different sources of hospitals under Ahvaz University, Iran. The isolates were examined by conventional microbiological methods and molecular test using PCR. The antibiotic susceptibility patterns of the isolates were determined by disk-diffusion and E-test. The biofilm formation ability of the isolates was investigated by Modified Congo red agar and microtiter plate techniques. The presence of virulence genes was examined using Multiplex-PCR method.

RESULTS: Out of 57 VREfm isolates, 63.15% of isolates were biofilm producers. The frequency of biofilm producing isolates from clinical specimens, colonized patients and environmental sources were 78.26%, 60%, and 42.85%, respectively. The prevalence of acm, esp and hyl genes among biofilm producing isolates was 86.10%, 55.56% and 52.77%, respectively. There was statistically significant association between esp gene and biofilm formation among isolates from the clinical specimens.

CONCLUSION: Clinical isolates producing biofilms showed a positive association with the presence of the esp. Our study further suggests that the link between virulence genes and biofilms is affected by the environmental context.},
}

@article {pmid32023304,
year = {2020},
author = {Siebert, C and Villers, C and Pavlou, G and Touquet, B and Yakandawala, N and Tardieux, I and Renesto, P},
title = {Francisella novicida and F. philomiragia biofilm features conditionning fitness in spring water and in presence of antibiotics.},
journal = {PloS one},
volume = {15},
number = {2},
pages = {e0228591},
doi = {10.1371/journal.pone.0228591},
pmid = {32023304},
issn = {1932-6203},
abstract = {Biofilms are currently considered as a predominant lifestyle of many bacteria in nature. While they promote survival of microbes, biofilms also potentially increase the threats to animal and public health in case of pathogenic species. They not only facilitate bacteria transmission and persistence, but also promote spreading of antibiotic resistance leading to chronic infections. In the case of Francisella tularensis, the causative agent of tularemia, biofilms have remained largely enigmatic. Here, applying live and static confocal microscopy, we report growth and ultrastructural organization of the biofilms formed in vitro by these microorganisms over the early transition from coccobacillary into coccoid shape during biofilm assembly. Using selective dispersing agents, we provided evidence for extracellular DNA (eDNA) being a major and conserved structural component of mature biofilms formed by both F. subsp. novicida and a human clinical isolate of F. philomiragia. We also observed a higher physical robustness of F. novicida biofilm as compared to F. philomiragia one, a feature likely promoted by specific polysaccharides. Further, F. novicida biofilms resisted significantly better to ciprofloxacin than their planktonic counterparts. Importantly, when grown in biofilms, both Francisella species survived longer in cold water as compared to free-living bacteria, a trait possibly associated with a gain in fitness in the natural aquatic environment. Overall, this study provides information on survival of Francisella when embedded with biofilms that should improve both the future management of biofilm-related infections and the design of effective strategies to tackle down the problematic issue of bacteria persistence in aquatic ecosystems.},
}

@article {pmid32021324,
year = {2020},
author = {Hu, L and Shi, Y and Xu, Q and Zhang, L and He, J and Jiang, Y and Liu, L and Leptihn, S and Yu, Y and Hua, X and Zhou, Z},
title = {Capsule Thickness, Not Biofilm Formation, Gives Rise to Mucoid Acinetobacter baumannii Phenotypes That are More Prevalent in Long-Term Infections: A Study of Clinical Isolates from a Hospital in China.},
journal = {Infection and drug resistance},
volume = {13},
number = {},
pages = {99-109},
doi = {10.2147/IDR.S230178},
pmid = {32021324},
issn = {1178-6973},
abstract = {Background: Acinetobacter baumannii is a nosocomial pathogen of critical importance due to the increasing numbers of antibiotic-resistant isolates. Colonies can have a smooth or matt appearance, but also exhibit slimy, mucoid growth, with the latter being increasingly isolated in patients in recent years.

Methods: We isolated 60 A. baumannii strains from altogether 56 patients and found that all patients were infected by mucoid strains, with four patients having also matt phenotypes in addition to the mucoid ones. The morphology of the colonies and capsules was observed. The antibiotics susceptibilities were tested, and the biofilm formation ability was determined by crystal violet staining. The whole-genome sequencing (WGS) was performed on all the strains, and then the core genome multilocus sequence typing (cgMLST) and drug resistance gene analysis were performed. Finally, a part of isolates were selected to test virulence in a Galleria mellonella model.

Results: We observed much larger capsules in the mucoid strains compared to the matt isolates. But the mucoid phenotype did not correlate with the amount of biofilm produced by the strain. Almost all mucus-type A. baumannii were multi-drug resistant isolates, containing various antibiotic resistance genes. The main ST types of mucoid-type A.baumannii were ST191 and ST195, of which ST191 isolates were more virulence, while ST195 isolates were weaker.

Conclusion: The mucoid A. baumannii had resistance to most antibiotics and some strains had high virulence, which should be paid attention in clinical.},
}

@article {pmid32021094,
year = {2020},
author = {Xie, Y and Liu, X and Zhou, P},
title = {In vitro Antifungal Effects of Berberine Against Candida spp. In Planktonic and Biofilm Conditions.},
journal = {Drug design, development and therapy},
volume = {14},
number = {},
pages = {87-101},
doi = {10.2147/DDDT.S230857},
pmid = {32021094},
issn = {1177-8881},
abstract = {Purpose: Antifungal resistance associated with the extensive use of antifungals and biofilm formation presents major clinical challenges. Thus, new therapeutic strategies for fungal infections are urgently required. This study aimed to evaluate the in vitro antifungal effects of the natural bioactive alkaloid berberine against Candida spp. in planktonic and biofilm conditions.

Methods: Using the CLSI M27-A3 reference method for broth dilution antifungal susceptibility testing of yeasts, the MICs for five standard strains comprised of Candida albicans (ATCC 10231, ATCC 90028), Candida krusei (ATCC 6258), Candida glabrata (ATCC 90030), Candida dubliniensis (MYA 646), and six clinical isolates (CLC1-CLC6) were tested. The 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay was used to evaluate the inhibitory effects of berberine against Candida biofilms. The optical density value at 490 nm was measured and illustrated using concentration-absorbance curves. Finally, the effects were quantified by confocal laser scanning microscopy (CLSM), and 3-dimensional reconstruction was performed. The viability inhibition rates, biofilm formation, and thickness decrease rates were tested and analyzed using independent-samples t-test. The differences among the five Candida strains were analyzed using one way ANOVA.

Results: The MICs for the five standard strains described above were 80, 160, 10, 20, and 40 μg/mL, respectively, which was similar to that of the clinical isolates, suggesting the stable, broad-spectrum antifungal activity of berberine. Berberine exerted concentration-dependent inhibitory effects against Candida biofilms, which were enhanced with the maturation of Candida biofilms. Berberine decreased the viability of Candida biofilms, with inhibition rates by CLSM ranging from 19.89 ± 0.57% to 96.93 ± 1.37%. Following 3-dimensional reconstruction, the biofilms of the berberine-treated group displayed a poorly developed architecture, and the biofilm thickness decrease rates ranged from 15.49 ± 8.45% to 30.30 ± 15.48%.

Conclusion: Berberine exhibited significant antifungal activity in Candida spp. The results provide a useful reference for multiple Candida infections and biofilm infections associated with antifungal resistance. Therefore, berberine might have novel therapeutic potential as an antifungal agent or a major active component of antifungal drugs.},
}

@article {pmid32020907,
year = {2020},
author = {Machineni, L},
title = {Effects of biotic and abiotic factors on biofilm growth dynamics and their heterogeneous response to antibiotic challenge.},
journal = {Journal of biosciences},
volume = {45},
number = {},
pages = {},
pmid = {32020907},
issn = {0973-7138},
abstract = {Over the last couple of decades, with the crisis of new antimicrobial arsenal, multidrug-resistant clinical pathogens have been observed extensively. In clinical and medical settings, these persistent pathogens predominantly grow as complex heterogeneous structures enmeshed in a self-produced exopolysaccharide matrix, termed as biofilms. Since biofilms can rapidly form by adapting new environmental surroundings and have potential effect on human health, it is critical to study them promptly and consistently. Biofilm infections are challenging in the contamination of medical devices and implantations, food processing and pharmaceutical industrial settings, and in dental area caries, periodontitis and so on. The persistence of infections associated with biofilms has been mainly attributed to the increased antibiotic resistance offered by the cells growing in biofilms. In fact, it is well known that this recalcitrance of bacterial biofilms is multifactorial, and there are several resistance mechanisms that may act in parallel in order to provide an enhanced level of resistance to the biofilm. In combination, distinct resistance mechanisms significantly decrease our ability to control and eradicate biofilm-associated infections with current antimicrobial arsenal. In addition, various factors are known to influence the process of biofilm formation, growth dynamics, and their heterogeneous response towards antibiotic therapy. The current review discusses the contribution of cellular and physiochemical factors on the growth dynamics of biofilm, especially their role in antibiotic resistance mechanisms of bacterial population living in surface attached growth mode. A systematic investigation on the effects and treatment of biofilms may pave the way for novel therapeutic strategies to prevent and treat biofilms in healthcare and industrial settings.},
}

@article {pmid32020424,
year = {2020},
author = {Çankirili, NK and Kart, D and Çelebi-Saltik, B},
title = {Evaluation of the biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa on human umbilical cord CD146+ stem cells and stem cell-based decellularized matrix.},
journal = {Cell and tissue banking},
volume = {},
number = {},
pages = {},
doi = {10.1007/s10561-020-09815-6},
pmid = {32020424},
issn = {1573-6814},
support = {TYL-2019-17802//Hacettepe University, Scientific Research Project Coordination Unit/ ; },
abstract = {This study aims to evaluate the CD146+ stem cells obtained from the human umbilical cord and their extracellular matrix proteins on in vitro Pseudomonas aeruginosa and Staphylococcus aureus biofilms to understand their possible antimicrobial activity. CD146+ stem cells were determined according to cell surface markers and differentiation capacity. Characterization of the decellularized matrix was done with DAPI, Masson's Trichrome staining and proteome analysis. Cell viability/proliferation of cells in co-cultures was evaluated by WST-1 and crystal-violet staining. The effects of cells and decellularized matrix proteins on biofilms were investigated on a drip flow biofilm reactor and their effects on gene expression were determined by RT-qPCR. We observed that CD146/105+ stem cells could differentiate adipogenically and decellularized matrix showed negative DAPI and positive collagen staining with Masson' s Trichrome. Proteome analysis of the decellularized matrix revealed some matrix components and growth factors. Although the decellularized matrix significantly reduced the cell counts of P. aeruginosa, no significant difference was observed for S. aureus cells in both groups. Supporting data was obtained from the gene expression results of P. aeruginosa with the significant down-regulation of rhlR and lasR. For S. aureus, icaADBC genes were significantly up-regulated when grown on the decellularized matrix.},
}

@article {pmid32018281,
year = {2020},
author = {Zavattini, A and Cowie, J and Niazi, S and Giovarruscio, M and Sauro, S and Foschi, F},
title = {Reduction of an in vitro Intraradicular Multispecies Biofilm Using Two Rotary Instrumentation Sequences.},
journal = {European journal of dentistry},
volume = {},
number = {},
pages = {},
doi = {10.1055/s-0040-1701541},
pmid = {32018281},
issn = {1305-7456},
abstract = {OBJECTIVE: The purpose of this research was to investigate the effect per se of two shaping and cleaning techniques on the reduction of an in vitro multispecies biofilm.

MATERIALS AND METHODS: A total of 39 freshly extracted monoradicular teeth for periodontal reason were decoronated. Roots were sectioned longitudinally. After autoclaving, a specific stressed biofilm was grown on the root halves that were subsequently reassembled in a silicone index. Two treatments (n = 9 each)-RaCe (Schottlander; Letchworth Garden City, United Kingdom) and ProTaper Gold (PTG; Dentsply Maillefer, Baillagues, Switzerland)-were tested; three noninstrumented samples served as a control group and three were rinsed with saline. Posttreatment samples were taken at three different levels of the root. Colony-forming units were counted after incubations. Additionally, three treatments (n = 5 each)-RaCe, PTG, and saline only-were evaluated under a confocal laser scanning microscope (CLSM). Statistical analysis was conducted using Tukey's test and analysis of variance to evaluate the post-instrumentation bioburden.

RESULTS: Both instrumentations were able to reduce the biofilm; however, differences were not present between them (p > 0.05). CLSM showed biofilm killing and disruption through mechanical shaping alone.

CONCLUSIONS: Intraradicular biofilm is reduced with mechanical shaping. There was no difference between RaCe and PTG systems in biofilm reduction despite differences in design, file sequence, and rotational speed.},
}

@article {pmid32017602,
year = {2020},
author = {Bumunang, E and Ateba, C and Stanford, K and McAllister, TA and Niu, YD},
title = {Biofilm formation by South-African Shiga toxigenic non-O157 Escherichia coli on stainless steel coupons.},
journal = {Canadian journal of microbiology},
volume = {},
number = {},
pages = {},
doi = {10.1139/cjm-2019-0554},
pmid = {32017602},
issn = {1480-3275},
abstract = {This study examined the biofilm-forming ability of six non-O157 STEC serotypes: O116:H21, wzx-Onovel5:H19, O129:H21, O129:H23, O26:H11 and O154:H10 on stainless steel coupons after 24, 48 and 72 h of incubation at 22 °C and 168 h at 10 °C. Based on crystal violet staining, O129:H23 and O154:H10 were able to form biofilms on both the submerged surface and the air-liquid interface of coupons whereas, O116:H21, wzx-Onovel5:H19, O129:H21 and O26:H11 formed biofilm only at the air-liquid interface. Viable cell counts and scanning electron microscopy showed that biofilm formation increased (p < 0.05) over time. The biofilm-forming ability of non-O157 STEC was strongest (p < 0.05) at 22 °C after 48 h incubation. The strongest biofilm former regardless of temperature was O129:H23. Generally, at 10 °C, weak to no biofilm was observed for isolates O154:H10, O116:H21, wzx-Onovel5:H19, O26:H11 and O129:H21 after 168 h. This study found that temperature affected biofilm-forming ability of non-O157 STEC strains. Overall, our data indicate that the high potential biofilm formation abilities of isolates at 22 °C, suggests that non-O157 STEC strains could colonize stainless steel within food-processing facilities. This could serve as a potential source of adulteration and promote the dissemination of these potential pathogens in food.},
}

@article {pmid32013708,
year = {2020},
author = {Kamble, E and Pardesi, K},
title = {Antibiotic Tolerance in Biofilm and Stationary-Phase Planktonic Cells of Staphylococcus aureus.},
journal = {Microbial drug resistance (Larchmont, N.Y.)},
volume = {},
number = {},
pages = {},
doi = {10.1089/mdr.2019.0425},
pmid = {32013708},
issn = {1931-8448},
abstract = {The ability of Staphylococcus aureus to form biofilms and persisters is a major cause of recalcitrant infections that are difficult to treat. We have examined time-dependent variation in persister population present in stationary-phase planktonic cells and biofilms of S. aureus when treated with bactericidal antibiotics having different cellular targets. Fourteen isolates identified as S. aureus were found to be resistant to three to nine classes of antibiotics tested according to the CLSI guidelines. Among the sensitive isolates, S48 was found to be the strongest biofilm producer, whereas J6 was the weakest. The four antibiotics, ciprofloxacin, daptomycin, tobramycin, and vancomycin, inhibited biofilm formation, whereas daptomycin was the strongest in disrupting 24-hr-old biofilm. Treatment of stationary-phase planktonic cells with 100 × minimum inhibitory concentration (MIC) of these antibiotics showed a typical biphasic pattern indicating the presence of persister cells. Twenty-four-hour-old biofilm of the two isolates tested at 100 × MIC of the antibiotics showed a similar biphasic pattern. Tolerance of biofilm cells was greater as compared with planktonic cells, which could be due to elevated number of persisters found in the biofilm as compared with planktonic cells.},
}

@article {pmid32013373,
year = {2020},
author = {Kirmusaoglu, S},
title = {Improved β-Lactam Susceptibility Against ica-Dependent Biofilm-Embedded Staphylococcus aureus by 2-Aminothiazole.},
journal = {Clinical laboratory},
volume = {66},
number = {1},
pages = {},
doi = {10.7754/Clin.Lab.2020.191112},
pmid = {32013373},
issn = {1433-6510},
abstract = {BACKGROUND: Due to the emergence of methicillin-resistant Staphylococcus aureus (MRSA) producing biofilm, causing recurrent infections worldwide, new therapeutic combinations need to be discovered to prevent resistance and to make treatment available. It was aimed to improve β-lactam susceptibility against ica-dependent biofilm-embedded Staphylococcus aureus (S. aureus), even in blaZ and mecA carriers, by 2-aminothiazole.

METHODS: Virulence genes in isolates were detected by qRT-PCR. MICs, MBCs of 2-aminothiazole and β-lactams against planktonic and sessile ica-dependent biofilm-producer isolates were investigated. Activities of 2-aminothiazole combined β-lactams against sessile one MRSA ATCC 43300, one MRSA, and two MSSA were investigated by checkerboard-assay.

RESULTS: Activities of 2-aminothiazole combined β-lactams were found synergistic and partially-synergistic against both biofilm-embedded MRSA and MSSA-isolates with FIC-indexes ranging between 0.193 - 0.387 and 0.535 - 0.745 and between 0.358 - 0.415 and 0.707 - 1.0, respectively. MICs of β-lactams against MSSA and MRSA were decreased 2- to 8-fold and 0- to 8-fold by sub-MICs of 2-aminothiazole, respectively.

CONCLUSIONS: Sub-MIC 2-aminothiazole combined Sub-MIC β-lactams can be a choice in varying applications to treat biofilm-associated staphylococcal infections.},
}

@article {pmid32013221,
year = {2020},
author = {Yuan, G and Li, P and Xu, X and Li, P and Zhong, Q and He, S and Yi, H and Yi, W and Guan, Y and Wen, ZT},
title = {Azalomycin F5a Eradicates Staphylococcus aureus Biofilm by Rapidly Penetrating and Subsequently Inducing Cell Lysis.},
journal = {International journal of molecular sciences},
volume = {21},
number = {3},
pages = {},
doi = {10.3390/ijms21030862},
pmid = {32013221},
issn = {1422-0067},
support = {81660578, 81960636 and 81460529//National Natural Science Foundation of China/ ; 20192ACBL20020//Natural Science Foundation of Jiangxi Province/ ; },
abstract = {Antimicrobial resistance has emerged as a serious threat to public health. Bacterial biofilm, as a natural lifestyle, is a major contributor to resistance to antimicrobials. Azalomycin F5a, a natural guanidine-containing polyhydroxy macrolide, has remarkable activities against Gram-positive bacteria, including Staphylococcus aureus, a major causative agent of hospital-acquired infections. To further evaluate its potential to be developed as a new antimicrobial agent, its influence on S. aureus biofilm formation was evaluated using the crystal violet method, and then its eradication effect against mature biofilms was determined by confocal laser scanning microscopy, the drop plate method, and regrowth experiments. The results showed that azalomycin F5a could significantly inhibit S. aureus biofilm formation, and such effects were concentration dependent. In addition, it can also eradicate S. aureus mature biofilms with the minimum biofilm eradication concentration of 32.0 μg/mL. As extracellular deoxyribonucleic acid (eDNA) plays important roles in the structural integrity of bacterial biofilm, its influence on the eDNA release in S. aureus biofilm was further analyzed using gel electrophoresis. Combined with our previous works, these results indicate that azalomycin F5a could rapidly penetrate biofilm and causes damages to the cell membrane, leading to an increase in DNase release and eventually eradicating S. aureus biofilm.},
}

@article {pmid32011777,
year = {2020},
author = {Davis, SC and Pastar, I},
title = {Reply to "Questioning the use of an acute porcine wound model to assess anti-biofilm activity of dressings".},
journal = {Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society},
volume = {},
number = {},
pages = {},
doi = {10.1111/wrr.12795},
pmid = {32011777},
issn = {1524-475X},
}

@article {pmid32011702,
year = {2020},
author = {Ceballos Garzon, A and Amado, D and Robert, E and Parra Giraldo, CM and Le Pape, P},
title = {Impact of calmodulin inhibition by fluphenazine on susceptibility, biofilm formation and pathogenicity of caspofungin-resistant Candida glabrata.},
journal = {The Journal of antimicrobial chemotherapy},
volume = {},
number = {},
pages = {},
doi = {10.1093/jac/dkz565},
pmid = {32011702},
issn = {1460-2091},
abstract = {BACKGROUND: In recent decades, Candida glabrata has emerged as a frequent cause of life-threatening fungal infection. In C. glabrata, echinocandin resistance is associated with mutations in FKS1/FKS2 (β-1,3-glucan synthase). The calmodulin/calcineurin pathway is implicated in response to antifungal stress and calcineurin gene disruption specifically reverses Fks2-mediated resistance of clinical isolates.

OBJECTIVES: We evaluated the impact of calmodulin inhibition by fluphenazine in two caspofungin-resistant C. glabrata isolates.

METHODS: C. glabrata isolates were identified by ITS1/ITS4 (where ITS stands for internal transcribed spacer) sequencing and the echinocandin target FKS1/FKS2 genes were sequenced. Susceptibility testing of caspofungin in the presence of fluphenazine was performed by a modified CLSI microbroth dilution method. The effect of the fluphenazine/caspofungin combination on heat stress (37°C or 40°C), oxidative stress (0.2 and 0.4 mM menadione) and biofilm formation (polyurethane catheter) was analysed. A Galleria mellonella model using blastospores (1 × 109 cfu/mL) was developed to evaluate the impact of this combination on larval survival.

RESULTS: F659del was found in the FKS2 gene of both resistant strains. In these clinical isolates, fluphenazine increased susceptibility to caspofungin and reduced their thermotolerance. Furthermore, the fluphenazine/caspofungin combination significantly impaired biofilm formation in an in vitro polyurethane catheter model. All these features participated in the increasing survival of infected G. mellonella after combination treatment in comparison with caspofungin alone.

CONCLUSIONS: In a repurposing strategy, our findings confirm that calmodulin could provide a relevant target in life-threatening fungal infectious diseases.},
}

@article {pmid32010109,
year = {2019},
author = {Hwang, SH and Park, JH and Lee, B and Choi, SH},
title = {A Regulatory Network Controls cabABC Expression Leading to Biofilm and Rugose Colony Development in Vibrio vulnificus.},
journal = {Frontiers in microbiology},
volume = {10},
number = {},
pages = {3063},
doi = {10.3389/fmicb.2019.03063},
pmid = {32010109},
issn = {1664-302X},
abstract = {Biofilms provide bacteria with protection from environmental stresses and host immune defenses. The pathogenic marine bacterium Vibrio vulnificus forms biofilms and colonizes environmental niches such as oysters. The cabABC operon encodes an extracellular matrix protein CabA and the corresponding type I secretion system, which are essential for biofilm and rugose colony development of V. vulnificus. In this study, molecular biological analyses revealed the roles of three transcriptional regulators BrpR, BrpT, and BrpS in the regulatory pathway for the cabABC operon. BrpR induces brpT and BrpT in turn activates the cabABC operon in a sequential cascade, contributing to development of robust biofilm structures. BrpT also activates brpS, but BrpS represses brpT, constituting a negative feedback loop that stabilizes brpT expression. BrpT and BrpS directly bind to specific sequences upstream of cabA, and they constitute a feedforward loop in which BrpT induces brpS and together with BrpS activates cabABC, leading to precise regulation of cabABC expression. Accordingly, BrpS as well as BrpT plays a crucial role in complete development of rugose colonies. This elaborate network of three transcriptional regulators BrpR, BrpT, and BrpS thus tightly controls cabABC regulation, and contributes to successful development of robust biofilms and rugose colonies in V. vulnificus.},
}

@article {pmid32010107,
year = {2019},
author = {Völkel, S and Hein, S and Benker, N and Pfeifer, F and Lenz, C and Losensky, G},
title = {How to Cope With Heavy Metal Ions: Cellular and Proteome-Level Stress Response to Divalent Copper and Nickel in Halobacterium salinarum R1 Planktonic and Biofilm Cells.},
journal = {Frontiers in microbiology},
volume = {10},
number = {},
pages = {3056},
doi = {10.3389/fmicb.2019.03056},
pmid = {32010107},
issn = {1664-302X},
abstract = {Halobacterium salinarum R1 is an extremely halophilic archaeon capable of adhesion and forming biofilms, allowing it to adjust to a range of growth conditions. We have recently shown that living in biofilms facilitates its survival under Cu2+ and Ni2+ stress, with specific rearrangements of the biofilm architecture observed following exposition. In this study, quantitative analyses were performed by SWATH mass spectrometry to determine the respective proteomes of planktonic and biofilm cells after exposition to Cu2+ and Ni2+.Quantitative data for 1180 proteins were obtained, corresponding to 46% of the predicted proteome. In planktonic cells, 234 of 1180 proteins showed significant abundance changes after metal ion treatment, of which 47% occurred in Cu2+ and Ni2+ treated samples. In biofilms, significant changes were detected for 52 proteins. Only three proteins changed under both conditions, suggesting metal-specific stress responses in biofilms. Deletion strains were generated to assess the potential role of selected target genes. Strongest effects were observed for ΔOE5245F and ΔOE2816F strains which exhibited increased and decreased biofilm mass after Ni2+ exposure, respectively. Moreover, EPS obviously plays a crucial role in H. salinarum metal ion resistance. Further efforts are required to elucidate the molecular basis and interplay of additional resistance mechanisms.},
}

@article {pmid32009731,
year = {2019},
author = {Merigo, E and Chevalier, M and Conti, S and Ciociola, T and Fornaini, C and Manfredi, M and Vescovi, P and Doglio, A},
title = {Antimicrobial effect on Candida albicans biofilm by application of different wavelengths and dyes and the synthetic killer decapeptide KP.},
journal = {Laser therapy},
volume = {28},
number = {3},
pages = {180-186},
doi = {10.5978/islsm.28_19-OR-14},
pmid = {32009731},
issn = {0898-5901},
abstract = {The aim of this study was to test the application in vitro of different laser wavelengths at a low fluence in combination or not with proper photosensitizing dyes on Candida albicans biofilm with or without a synthetic killer decapeptide (KP). Candida albicans SC5314 was grown on Sabouraud dextrose agar plates at 37°C for 24 h. Cells were suspended in RPMI 1640 buffered with MOPS and cultured directly on the flat bottom of 96-wells plates. The previously described killer decapeptide KP was used in this study. Three different combinations of wavelengths and dyes were applied, laser irradiation has been performed at a fluence of 10 J/cm2. The effect on C. albicans biofilm was evaluated by the XTT assay. Microscopic observations were realized by fluorescence optic microscopy with calcofluor white and propidium iodide. Compared with control, no inhibition of C. albicans biofilm viability was obtained with application of red, blue and green lasers alone or with any combination of red diode laser, toluidine blue and KP. The combined application of blue diode laser with curcumin and/or KP showed always a very significant inhibition, as curcumin alone and the combination of curcumin and KP did, while combination of blue diode laser and KP gave a less significant inhibition, the same obtained with KP alone. The combined application of green diode laser with erythrosine and/or KP showed always a very significant inhibition, as the combination of erythrosine and KP did, but no difference was observed with respect to the treatment with erythrosine alone. Again, combination of green diode laser and KP gave a significant inhibition, although paradoxically lower than the one obtained with KP alone. Treatment with KP alone, while reducing biofilm viability did not cause C. albicans death in the adopted experimental conditions. On the contrary, combined treatment with blue laser, curcumin and KP, as well as green laser, erythrosine and KP led to death most C. albicans cells. The combination of laser light at a fluence of 10 J/cm2 and the appropriate photosensitizing agent, together with the use of KP, proved to exert differential effects on C. albicans biofilm.},
}

@article {pmid32009660,
year = {2020},
author = {Palmer, MB and Wang, Y and White, AP},
title = {Discovering CsgD Regulatory Targets in Salmonella Biofilm Using Chromatin Immunoprecipitation and High-Throughput Sequencing (ChIP-seq).},
journal = {Journal of visualized experiments : JoVE},
volume = {},
number = {155},
pages = {},
doi = {10.3791/60736},
pmid = {32009660},
issn = {1940-087X},
abstract = {Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a technique that can be used to discover the regulatory targets of transcription factors, histone modifications, and other DNA-associated proteins. ChIP-seq data can also be used to find differential binding of transcription factors in different environmental conditions or cell types. Initially, ChIP was performed through hybridization on a microarray (ChIP-chip); however, ChIP-seq has become the preferred method through technological advancements, decreasing financial barriers to sequencing, and massive amounts of high-quality data output. Techniques of performing ChIP-seq with bacterial biofilms, a major source of persistent and chronic infections, are described in this protocol. ChIP-seq is performed on Salmonella enterica serovar Typhimurium biofilm and planktonic cells, targeting the master biofilm regulator, CsgD, to determine differential binding in the two cell types. Here, we demonstrate the appropriate amount of biofilm to harvest, normalizing to a planktonic control sample, homogenizing biofilm for cross-linker access, and performing routine ChIP-seq steps to obtain high quality sequencing results.},
}

@article {pmid32009221,
year = {2020},
author = {Schubiger, CB and Hoang, KHT and Häse, CC},
title = {Sodium antiporters of Pseudomonas aeruginosa in challenging conditions: effects on growth, biofilm formation, and swarming motility.},
journal = {Journal, genetic engineering & biotechnology},
volume = {18},
number = {1},
pages = {4},
doi = {10.1186/s43141-020-0019-y},
pmid = {32009221},
issn = {2090-5920},
abstract = {BACKGROUND: Pseudomonas aeruginosa is a bacterial pathogen that can cause grave and sometimes chronic infections in patients with weakened immune systems and cystic fibrosis. It is expected that sodium/proton transporters in the cellular membrane are crucial for the organism's survival and growth under certain conditions, since many cellular processes rely on the maintenance of Na+ and H+ transmembrane gradients.

RESULTS: This study focused on the role of the primary and secondary proton and/or sodium pumps Mrp, Nuo, NhaB, NhaP, and NQR for growth, biofilm formation, and swarming motility in P. aeruginosa. Using mutants with gene deletions, we investigated the impact of each sodium pump's absence on the overall growth, biofilm formation, motility, and weak acid tolerance of the organism. We found that the absence of some, but not all, of the sodium pumps have a deleterious effect on the different phenotypes of P. aeruginosa.

CONCLUSION: The absence of the Mrp sodium/proton antiporter was clearly important in the organism's ability to survive and function in environments of higher pH and sodium concentrations, while the absence of Complex I, which is encoded by the nuo genes, had some consistent impact on the organism's growth regardless of the pH and sodium concentration of the environment.},
}

@article {pmid32008118,
year = {2020},
author = {Jin, X and Marshall, JS and Wargo, MJ},
title = {Hybrid Model of Bacterial Biofilm Growth.},
journal = {Bulletin of mathematical biology},
volume = {82},
number = {2},
pages = {27},
doi = {10.1007/s11538-020-00701-6},
pmid = {32008118},
issn = {1522-9602},
support = {NNX16AQ96A/NASA/NASA/United States ; },
abstract = {Bacterial biofilms play a critical role in environmental processes, water treatment, human health, and food processing. They exhibit highly complex dynamics due to the interactions between the bacteria and the extracellular polymeric substance (EPS), water, and nutrients and minerals that make up the biofilm. We present a hybrid computational model in which the dynamics of discrete bacterial cells are simulated within a multiphase continuum, consisting of EPS and water as separate interacting phases, through which nutrients and minerals diffuse. Bacterial cells in our model consume water and nutrients in order to grow, divide, and produce EPS. Consequently, EPS flows outward from the bacterial colony, while water flows inward. The model predicts bacterial colony formation as a treelike structure. The distribution of bacterial growth and EPS production is found to be sensitive to the pore spacing between bacteria and the consumption of nutrients within the bacterial colony. Forces that are sometimes neglected in biofilm simulations, such as lubrication force between nearby bacterial cells and osmotic (swelling) pressure force resulting from gradients in EPS concentration, are observed to have an important effect on biofilm growth via their influence on bacteria pore spacing and associated water/nutrient percolation into the bacterial colony.},
}