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Bibliography on: Biofilm

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ESP: PubMed Auto Bibliography 20 Nov 2019 at 01:33 Created: 


Wikipedia: Biofilm A biofilm is any group of microorganisms in which cells stick to each other and often also to a surface. These adherent cells become embedded within a slimy extracellular matrix that is composed of extracellular polymeric substances (EPS). The EPS components are produced by the cells within the biofilm and are typically a polymeric conglomeration of extracellular DNA, proteins, and polysaccharides. Because they have three-dimensional structure and represent a community lifestyle for microorganisms, biofilms are frequently described metaphorically as cities for microbes. Biofilms may form on living or non-living surfaces and can be prevalent in natural, industrial and hospital settings. The microbial cells growing in a biofilm are physiologically distinct from planktonic cells of the same organism, which, by contrast, are single-cells that may float or swim in a liquid medium. Biofilms can be present on the teeth of most animals as dental plaque, where they may cause tooth decay and gum disease. Microbes form a biofilm in response to many factors, which may include cellular recognition of specific or non-specific attachment sites on a surface, nutritional cues, or in some cases, by exposure of planktonic cells to sub-inhibitory concentrations of antibiotics. When a cell switches to the biofilm mode of growth, it undergoes a phenotypic shift in behavior in which large suites of genes are differentially regulated.

Created with PubMed® Query: biofilm[title] NOT pmcbook NOT ispreviousversion

Citations The Papers (from PubMed®)

RevDate: 2019-11-19

Godovalov AP, TI Karpunina (2019)

[The determination of biofilm composition of gram-positive bacteria.].

Klinicheskaia laboratornaia diagnostika, 64(10):632-634.

Current methods of biofilm imaging do not support a differentiated assessment of its composition, since it is not possible to establish a substrate stained with crystal violet, as this dye can form complexes with both intracellular and extracellular structures. This approach does not adequately assess the anti-biofilm effects of drugs, while the results of studying the interaction of drugs with biofilm components can ensure their most correct choice. The aim of investigation was to study the possibility of applying the original modification of the current method to determine the ratio of the cellular part and the matrix of biofilms of gram-positive microorganisms. The biofilm components were analyzed using a two-step approach, when prepared biofilms of gram-positive microorganisms were stained with crystal violet for 5 minutes, followed by fixing the dye in bacterial cells with iodine solution, and then the colored products were dissolved with 95% alcohol: matrix components for 1 minute, total biofilm for 15 minutes, after which the composition of biofilms was estimated by the formula: M=(OP1/OP15)×100, Kb=100-M, where M is the proportion of the matrix,%; Kb - the proportion of the cellular component,%; OP1 - optical density of samples, when alcohol was allowed to dissolve the colored product for no more than 1 minute; OP15 - was the optical density of samples, when alcohol is allowed to dissolve the colored product for 15 minutes. It was shown that in the composition of the biofilm formed by the collection strain, the proportion of the matrix was 13.2%, and the cellular component accounted for 86.8%. When the same strain cultivated in the presence of an antibiotic, an increase in the biofilm matrix was observed, which is probably due to the compensatory response of the microorganism to the action of the antibiotic. The proposed approach to the study of biofilms makes it possible to evaluate its component composition. Obtaining additional information in this way can provide, inter alia, an increase in the effectiveness of antimicrobial therapy while reducing the study time.

RevDate: 2019-11-19

Rahdar HA, Malekabad ES, Dadashi AR, et al (2019)

Correlation between biofilm formation and carbapenem resistance among clinical isolates of Klebsiella pneumoniae.

Ethiopian journal of health sciences, 29(6):745-750.

Background: Klebsiella pneumoniae is a Gram-negative enteric bacterium that causes nosocomial infections; this bacterium has survived from harsh condition using biofilm formation in hospital equipment and cause severe infection. In the other hand, the emergence and extension of carbapenem resistance burden among K. pneumonia producing biofilm is the current concern of public health services. There are controversial findings about this subject. The aim of this study was to evaluate the correlation between biofilm formation and resistance to carbapenem among clinical isolates of K. pneumoniae.

Methods: A total of 160 K. pneumoniae isolates were collected from various infections of hospitalized patients. The Carba NP test and molecular methods were used for detection of carbapenem resistance isolates of K. pneumonia. Subsequently, the ability for biofilm production was performed from all isolates. Finally, Correlation of biofilm formation among carbapenem resistant isolates was calculated using χ2 and Fisher's exact tests.

Results: Among K. pneumoniae isolates 42.5% have carbapenemase activity by Carba NP test, while carbapenemase genes were detected in 35.6% of isolates in amplification assay. Moreover, there are 52.5% (n= 84) of all isolates were formed a strong biofilm, while 38.1% (n= 61) and 9.3% (n= 15) of isolates were middle and weak biofilm producer, respectively. Among carbapenem resistant cases (n= 68), there are 77.9% (n= 53) and 22% (n= 15) of isolates were reported as strong and middle biofilm producer, respectively. We see a significant correlation was seen between biofilm formation ability and carbapenem resistant isolates (p-value < 0.00001).

Conclusion: The increase of carbapenem resistance burden in biofilm producing isolates of K. pneumoniae is considered as serious alert and the basic measures to combat this phenomenon is imperative.

RevDate: 2019-11-19

Liu Y, Feng H, Chen L, et al (2019)

Root-secreted spermine binds to Bacillus amyloliquefaciens SQR9 histidine kinase KinD and modulates biofilm formation.

Molecular plant-microbe interactions : MPMI [Epub ahead of print].

The signal molecules in root exudates that are sensed by plant growth-promoting rhizobacteria (PGPRs) are critical to regulate their root colonization. Phosphorylated Spo0A is an important global transcriptional regulator that controls colonization and sporulation in Bacillus species. In this study, we found that deletion of kinD from PGPR strain Bacillus amyloliquefaciens SQR9, encoding an original phosphate donor of Spo0A, resulted in reduced biofilm formation in root exudates compared with the wild type strain, indicating that KinD is responsible for sensing root exudates. Ligands of B. amyloliquefaciens SQR9 KinD in cucumber root exudates were determined by both the non-targeted ligand fishing method and the targeted surface plasmon resonance (SPR) detection method. In total, we screened 80 compounds in root exudates for binding to KinD and found that spermine and guanosine could bind to KinD with KD values of 213 μM and 51 μΜ, respectively. In addition, calcium L-threonate, N-acetyl-L-aspartic acid, sodium decanoic acid and parabanic acid could also bind to KinD weakly. Then, the 3-dimensional binding models were constructed to demonstrate the interactions between the root secreted signals and KinD. It was observed that exogenous spermine reduced the wrinkles of biofilm when kinD was defected, indicating that KinD might be involved in sensing root-secreted spermine and stabilizing biofilm in response to this negative effector. This study provided a new insight of interaction between rhizobacterial sensor and root secreted signals.

RevDate: 2019-11-19

Fernández-Calderón MC, Romero-Guzmán D, Ferrández-Montero A, et al (2019)

Impact of PLA/Mg films degradation on surface physical properties and biofilm survival.

Colloids and surfaces. B, Biointerfaces pii:S0927-7765(19)30761-1 [Epub ahead of print].

New biocompatible and bioabsorbable materials are currently being developed for bone regeneration. These serve as scaffolding for controlled drug release and prevent bacterial infections. Films of polylactic acid (PLA) polymers that are Mg-reinforced have demonstrated they have suitable properties and bioactive behavior for promoting the osseointegration process. However little attention has been paid to studying whether the degradation process can alter the adhesive physical properties of the biodegradable film and whether this can modify the biofilm formation capacity of pathogens. Moreover, considering that the concentration of Mg and other corrosion products may not be constant during the degradation process, the question that arises is whether these changes can have negative consequences in terms of the bacterial colonization of surfaces. Bacteria are able to react differently to the same compound, depending on its concentration in the medium and can even become stronger when threatened. In this context, physical surface parameters such as hydrophobicity, surface tension and zeta potential of PLA films reinforced with 10% Mg have been determined before and after degradation, as well as the biofilm formation capacity of Staphylococcus epidermidis. The addition of Mg to the films makes them less hydrophobic and the degradation also reduces the hydrophobicity and increases the negative charge of the surface, especially over long periods of time. Early biofilm formation at 8 h is consistent with the physical properties of the films, where we can observe a reduction in the bacterial biofilm formation. However, after 24 h of incubation, the biofilm formation increases significantly on the PLA/Mg films with respect to PLA control. The explosive release of Mg ions and other corrosion products within the first hours were not enough to prevent a greater biofilm formation after this initial time. Consequently, the Mg addition to the polymer matrix had a bacteriostatic effect but not a bactericidal one. Future works should aim to optimize the design and biofunctionality of these promising bioabsorbable composites for a degradation period suitable for the intended application.

RevDate: 2019-11-19

Pan J, Hu J, Liu B, et al (2019)

Enhanced quorum sensing of anode biofilm for better sensing linearity and recovery capability of microbial fuel cell toxicity sensor.

Environmental research pii:S0013-9351(19)30703-0 [Epub ahead of print].

MFC toxicity sensor has major hindrances that limit its practical application, such as the poor concentration-response relationship and inferior recovery capability after high toxicity shock. Till now, the direct influence of intrinsic properties on the performance of MFC toxicity sensor has not been well understood. Quorum sensing (QS) is a cell-to-cell communication strategy that indirectly affects the intrinsic properties of electroactive biofilms. In this work, commercially available QS autoinducers (AHLs) were applied to MFC toxicity sensor to manipulate anode biofilm for better sensing performance. The results showed that the addition of AHLs (C6-HSL, 3-OXO-C12-HSL) led to higher sensing linearity to a wider range of Pb2+. The voltage of MFC sensors with AHLs addition fully recovered even after 10 mg/L Cu2+ shock, indicating an enhanced recovery capability of MFC toxicity sensor. It was found that higher live/dead cells ratio and increased exoelectrogen Geobacter abundance were responsible for the superior sensing linearity and recovery capability of MFC toxicity sensor. Our work presented a novel and effective way to advance the process of MFC toxicity sensor application from the perspective of EABs.

RevDate: 2019-11-19

Ré ACS, Ferreira MP, Freitas O, et al (2019)

Antimicrobial effect of a local release system containing metronidazole against a Porphyromonas gingivalis biofilm.

Die Pharmazie, 74(11):665-666.

The aim of this study was to evaluate a semi-solid system containing metronidazole (MDZ) in presence of challenging conditions for drug release, as well its antimicrobial effect against Porphyromonas gingivalis biofilm. Biofilms grown in culture medium were exposed to a formulation containing MDZ or its vehicle. After 24, 48, and 72 h, biofilm viability were analyzed while MDZ was quantified in culture medium and buffer solution (control). MDZ formulation reduced bacterial viability when compared to control groups. The vehicle formulation also affected bacterial viability in relation to control at all periods. Culture medium impaired MDZ release compared to buffer solution at 24 h. The semi-solid system reported herein is able to release MDZ and maintain its levels at concentrations that control viability of P. gingivalis in 1- to 3-day-old biofilms.

RevDate: 2019-11-18

Zhang Z, Du W, Wang M, et al (2019)

Contribution of the colicin receptor CirA to biofilm formation, antibotic resistance, and pathogenicity of Salmonella Enteritidis.

Journal of basic microbiology [Epub ahead of print].

Salmonella Enteritidis is an important foodborne pathogen that can infect a wide range of animal species including human beings, resulting in great losses to commercial husbandry and human health. CirA is an outer membrane receptor involved in iron uptake and colicin1A/B-mediated competitive killing. Although iron uptake is crucial to bacterial virulence, limited literature is available about the role of CirA in infection. In the present work, we aimed to evaluate the role of CirA during S. Enteritidis infection. For this purpose, we generated a CirA-deficient mutant of the S. Enteritidis strain C50336 and examined its biological characteristics. The results showed that cirA gene inactivation caused sharply decreased biofilm formation and apparently impaired antibiotic resistance. Furthermore, the cirA gene deletion mutant showed markedly reduced adhesion and invasion to human epithelial cell line Caco-2 cells and decreased proliferation in mouse macrophage cell line RAW264.7 cells. Moreover, attenuated virulence was determined by a mouse model, with an LD50 increase of approximately 1,000-fold. These data indicated that CirA plays critical roles in the S. Enteritidis infection process.

RevDate: 2019-11-18

Qvortrup K, Hultqvist LD, Nilsson M, et al (2019)

Small Molecule Anti-biofilm Agents Developed on the Basis of Mechanistic Understanding of Biofilm Formation.

Frontiers in chemistry, 7:742.

Microbial biofilms are the cause of persistent infections associated with various medical implants and distinct body sites such as the urinary tract, lungs, and wounds. Compared with their free living counterparts, bacteria in biofilms display a highly increased resistance to immune system activities and antibiotic treatment. Therefore, biofilm infections are difficult or impossible to treat with our current armory of antibiotics. The challenges associated with biofilm infections have urged researchers to pursue a better understanding of the molecular mechanisms that are involved in the formation and dispersal of biofilms, and this has led to the identification of several steps that could be targeted in order to eradicate these challenging infections. Here we describe mechanisms that are involved in the regulation of biofilm development in Pseudomonas aeruginosa, Escherichia coli, and Acinetobacter baumannii, and provide examples of chemical compounds that have been developed to specifically inhibit these processes. These compounds include (i) pilicides and curlicides which inhibit the initial steps of biofilm formation by E. coli; (ii) compounds that interfere with c-di-GMP signaling in P. aeruginosa and E. coli; and (iii) compounds that inhibit quorum-sensing in P. aeruginosa and A. baumannii. In cases where compound series have a defined molecular target, we focus on elucidating structure activity relationship (SAR) trends within the particular compound series.

RevDate: 2019-11-18

Roussin M, Rabarioelina S, Cluzeau L, et al (2019)

Identification of a Contact-Dependent Growth Inhibition (CDI) System That Reduces Biofilm Formation and Host Cell Adhesion of Acinetobacter baumannii DSM30011 Strain.

Frontiers in microbiology, 10:2450.

Acinetobacter baumannii is a multidrug-resistant nosocomial opportunistic pathogen that is becoming a major health threat worldwide. In this study, we have focused on the A. baumannii DSM30011 strain, an environmental isolate that retains many virulence-associated traits. We found that its genome contains two loci encoding for contact-dependent growth inhibition (CDI) systems. These systems serve to kill or inhibit the growth of non-sibling bacteria by delivering toxins into the cytoplasm of target cells, thereby conferring the host strain a significant competitive advantage. We show that one of the two toxins functions as a DNA-damaging enzyme, capable of inducing DNA double-stranded breaks to the chromosome of Escherichia coli strain. The second toxin has unknown catalytic activity but stops the growth of E. coli without bactericidal effect. In our conditions, only one of the CDI systems was highly expressed in the A. baumannii DSM30011 strain and was found to mediate interbacterial competition. Surprisingly, the absence of this CDI system promotes adhesion of A. baumannii DSM30011 to both abiotic and biotic surfaces, a phenotype that differs from previously described CDI systems. Our results suggest that a specific regulation mediated by this A. baumannii DSM30011 CDI system may result in changes in bacterial physiology that repress host cell adhesion and biofilm formation.

RevDate: 2019-11-17

Gani KM, Nazir FU, Kumari S, et al (2019)

Role of treatment configuration in simultaneous removal of priority phthalic acid esters and nitrogen in a post anoxic integrated biofilm activated sludge system.

The Science of the total environment, 702:134733 pii:S0048-9697(19)34724-2 [Epub ahead of print].

To develop future wastewater treatment systems, focus is to improve/investigate existing biological wastewater treatment processes for the concurrent treatment of conventional pollution parameters (essentially nitrogen) and micro pollutants. Majority of the existing biological wastewater treatment systems were not designed for the removal of micro pollutants. This study focuses on understanding the role of treatment configuration for efficient removal of nitrogen and priority phthalic acid esters (PAEs) from real municipal wastewater in an integrated biofilm activated sludge (IBAS) system. The reactor was operated in two phases: Run I, without external carbon source in anoxic reactor and Run II, a nitrogen removal process, with partial diversion of untreated wastewater in anoxic reactor. Nitrogen removal was 70 ± 12% in both operational phases, however, during Run I, removal of PAEs fluctuated with an average removal of 60-78%. Comparatively, removal of PAEs in Run II varied over a smaller range with average removal increased to 89-95%. In both operational scenarios, secondary oxic tank contributed maximum to the overall removal of PAEs in treatment system. Mass balance calculations showed significant contribution of biodegradation towards overall removal of PAEs which was enhanced by the supply of external carbon source. Kinetics and model output supported the PAEs removal performance observed in different reaction environments of IBAS process. A correlation between food to microorganism (F/M) ratio and PAEs removal showed increase in PAEs removal with decrease in F/M ratio. The study showed that treatment configuration and F/M ratio may be one of the guiding parameters for efficient removal of PAEs in biological wastewater treatment.

RevDate: 2019-11-17

Wang SH, Chen CC, Lee CH, et al (2019)

Fungicidal and anti-biofilm activities of trimethylchitosan-stabilized silver nanoparticles against Candida species in zebrafish embryos.

International journal of biological macromolecules pii:S0141-8130(19)33986-8 [Epub ahead of print].

Herein, positively surface-charged silver nanoparticles (AgNPs) capped with trimethylchitosan nitrate (TMCN) were synthesized using an environmentally friendly method. Nano-sized TMCN-AgNPs (∼80 nm) with high zeta potential (>30 mV) provide sufficient static repulsion to stabilize colloid AgNPs in aqueous solutions without aggregation for more than 3 months. In in vitro cell cycle assays, TMCN-AgNPs showed low cytotoxicity towards L929 cells. A microdilution inhibition assay demonstrated the antifungal potential of TMCN-AgNPs, with a minimum inhibitory concentration of 0.06 mM against Candida tropicalis ATCC 750, and 0.46 mM against both Candida albicans ATCC 76615 and Candida glabrata ATCC 15545. Moreover, the addition of TMCN-AgNPs at 0.23 mM significantly reduced biofilm formation in 96-well plates with C. albicans and C. tropicalis. Importantly, when zebrafish eggs were infected with Candida cells, 0.23 mM TMCN-AgNPs greatly diminished the amount of biofilm on eggs and rescued the survival of embryos by up to 70%.

RevDate: 2019-11-17

Barnier C, Clerissi C, Lami R, et al (2019)

Description of Palleronia rufa sp. nov., a biofilm-forming and AHL-producing Rhodobacteraceae, reclassification of Hwanghaeicola aestuarii as Palleronia aestuarii comb. nov., Maribius pontilimi as Palleronia pontilimi comb. nov., Maribius salinus as Palleronia salina comb. nov., Maribius pelagius as Palleronia pelagia comb. nov. and emended description of the genus Palleronia.

Systematic and applied microbiology pii:S0723-2020(19)30313-3 [Epub ahead of print].

Strain MOLA 401T was isolated from marine waters in the southwest lagoon of New Caledonia and was shown previously to produce an unusual diversity of quorum sensing signaling molecules. This strain was Gram-negative, formed non-motile cocci and colonies were caramel. Optimum growth conditions were 30°C, pH 8 and 3% NaCl (w/v). Based on 16S rRNA gene sequence analysis, this strain was found to be closely related to Pseudomaribius aestuariivivens NBRC 113039T (96.9% of similarity), Maribius pontilimi DSM 104950T (96.4% of similarity) and Palleronia marisminoris LMG 22959T (96.3% of similarity), belonging to the Roseobacter group within the family Rhodobacteraceae. As its closest relatives, strain MOLA 401T is able to form a biofilm on polystyrene, supporting the view of Roseobacter group strains as prolific surface colonizers. An in-depth genomic study allowed us to affiliate strain MOLA 401T as a new species of genus Palleronia and to reaffiliate some of its closest relatives in this genus. Consequently, we describe strain MOLA 401T (DSM 106827T=CIP 111607T=BBCC 401T) for which we propose the name Palleronia rufa sp. nov. We also propose to emend the description of the genus Palleronia and to reclassify Maribius and Hwanghaeicola species as Palleronia species.

RevDate: 2019-11-16

Sun Z, Ding C, Xi J, et al (2019)

Enhancing biofilm formation in biofilters for benzene, toluene, ethylbenzene, and xylene removal by modifying the packing material surface.

Bioresource technology pii:S0960-8524(19)31565-2 [Epub ahead of print].

Polyurethane (PU) sponges are popular packing material in biofilters and their smooth and hydrophobic surface often leads to an uneven distribution and detachment of biofilms. In this work, the surface of PU sponge was modified to obtain higher roughness and positive charge. The performances of two biofilters (BF1 with pristine sponge and BF2 with modified sponge) for benzene, toluene, ethylbenzene, and xylene (BTEX) removal were investigated. Total Volatile Organic Compound (TVOC) removal efficiency and CO2 increment were 61% and 804 ppm for BF2 respectively after start-up, compared with 51% and 538 ppm for BF1. Analysis on biofilms showed that the modification of PU sponge significantly improved the microbial growth, viability and adhesive strength in biofilms, reduced extracellular polymeric substance (EPS) and changed the microbial community. These results demonstrate that modified sponge can enhance biofilm formation and BTEX removal in biofilters and may applied in large-scale applications.

RevDate: 2019-11-16

Choi HW, Park SY, Kang MK, et al (2019)

Comparative Analysis of Biofilm Removal Efficacy by Multisonic Ultracleaning System and Passive Ultrasonic Activation.

Materials (Basel, Switzerland), 12(21): pii:ma12213492.

The purpose of this study was to compare disinfection and the biofilm removal efficacy of the GentleWave System (Sonendo, Inc., Laguna Hills, CA, USA) with passive ultrasonic activation method. Forty-seven freshly extracted human molars were inoculated with Enterococcus faecalis and cultured for five weeks to establish biofilm. Eight molars were tested for confirmation of infection. Four of the eight teeth were not inoculated in order to provide a negative control. The remaining 39 inoculated molars were randomly separated into three treatment groups (n = 13 per group): Group 1-no treatment, Group 2-conventional rotary instrumentation and passive ultrasonic activation, and Group 3-minimal instrumentation and the GentleWave System treatment. Roots were subsequently prepared per standard histological tissue processing procedures. Modified Brown and Brenn stained sections and Hematoxylin and Eosin stained sections were visualized at 4× and 13.5× magnification using a stereomicroscope. The sections were scored and blindly analyzed by two independent evaluators, including a histopathologist, to evaluate the presence of biofilm on canal wall. A significant difference was found between Group 2 and Group 3 in both apical and middle regions (p = 0.001) of the mesial roots of mandibular molars and mesiobuccal roots of maxillary molars. Group 3 revealed significantly less biofilm than the controls (p = 0.003). The GentleWave System demonstrated significantly greater reduction in biofilm within the mesial roots of mandibular molars and mesiobuccal roots of maxillary molars than those treated with conventional rotary instrumentation and passive ultrasonic activation protocol.

RevDate: 2019-11-15

Azimi L, AR Lari (2019)

Colistin-resistant Pseudomonas aeruginosa clinical strains with defective biofilm formation.

GMS hygiene and infection control, 14:Doc12 pii:Doc12.

Aim: Colistin is the only effective antibiotic in some cases of Pseudomonasaeruginosa resistance to all tested antibiotics, even carbapenem. On the other hand, biofilm formation is one of the antibiotic resistance mechanisms in this bacterium. The aim of this study was to examine biofilm formation in colistin-resistant P. aeruginosa for the first time. Method: Two groups of P. aeruginosa were included in this study: 1) colistin-resistant and 2) colistin-susceptible. Biofilm formation was determined in these groups using the micro-tube test well as PCR to detect the genes involved in biofilm formation (ppk and molA). The plasmids for colistin resistance, mcr-1 and mcr-2, were also determined. P. aeruginosa ATCC 27853 was used as a control for all tests. Results: Strong biofilm formation was observed only in colistin-susceptible strains, and ppk and modA were not detected in colistin-resistant strains. The control strain P. aeruginosa ATCC 27853 possesses ppk and modA and is categorized as a strong biofilm formation group. According to the results of this study, colistin resistance is associated with defective biofilm formation, as reported by other studies on Acinetobacter baumannii.

RevDate: 2019-11-15

Penesyan A, Nagy SS, Kjelleberg S, et al (2019)

Rapid microevolution of biofilm cells in response to antibiotics.

NPJ biofilms and microbiomes, 5:34 pii:108.

Infections caused by Acinetobacter baumannii are increasingly antibiotic resistant, generating a significant public health problem. Like many bacteria, A. baumannii adopts a biofilm lifestyle that enhances its antibiotic resistance and environmental resilience. Biofilms represent the predominant mode of microbial life, but research into antibiotic resistance has mainly focused on planktonic cells. We investigated the dynamics of A. baumannii biofilms in the presence of antibiotics. A 3-day exposure of A. baumannii biofilms to sub-inhibitory concentrations of antibiotics had a profound effect, increasing biofilm formation and antibiotic resistance in the majority of biofilm dispersal isolates. Cells dispersing from biofilms were genome sequenced to identify mutations accumulating in their genomes, and network analysis linked these mutations to their phenotypes. Transcriptomics of biofilms confirmed the network analysis results, revealing novel gene functions of relevance to both resistance and biofilm formation. This approach is a rapid and objective tool for investigating resistance dynamics of biofilms.

RevDate: 2019-11-15

Katoch P, Gupta K, Yennamalli RM, et al (2019)

Random insertion transposon mutagenesis of Mycobacterium fortuitum identified mutant defective in biofilm formation.

Biochemical and biophysical research communications pii:S0006-291X(19)32141-2 [Epub ahead of print].

Mycobacterium fortuitum has emerged as a nosocomial infectious agent and biofilm formation attributed for the presence of this bacterium in hospital environment. Transposon random mutagenesis was used to identify membrane-proteins for biofilm formation in M. fortuitum. Ten mutants were shortlisted from a library of 450 mutants for examine their biofilm forming ability. Comparative biofilm ability with respect to wild type M. fortuitum ATCC 6841 showed an altered and delayed biofilm formation in one mutant namely, MT721. Sequence analysis revealed mutation in anthranilate phosphoribosyl transferase (MftrpD), which is associated with tryptophan operon. Functional interaction study of TrpD protein through STRING showed its interaction with chorismate utilizing proteins, majorly involved in synthesis of aromatic amino acid and folic acid, suggesting that biofilm establishment and maintenance requires components of central metabolism. Our study indicates important role of MftrpD in establishment and maintenance of biofilm by M. fortuitum, which may further be explored for drug discovery studies against mycobacterial infections.

RevDate: 2019-11-14

Bernardi S, Karygianni L, Filippi A, et al (2019)

Combining culture and culture-independent methods reveals new microbial composition of halitosis patients' tongue biofilm.

MicrobiologyOpen [Epub ahead of print].

BACKGROUND: Oral malodor is a very discomforting condition deriving from the presence of volatile sulfur compounds in the expired air. In halitosis of intraoral etiology, the volatile sulfur compounds are metabolic products of the oral microorganisms within the biofilm coating the tongue dorsum as well as other tissues in the oral cavity. The aim of this study was to characterize and compare the microbial composition of tongue biofilm in volunteers suffering from halitosis and healthy volunteers by means of both the culture method and culture-independent cloning technique.

RESULTS: A high bacterial variety (more than 80 different species) was detected using the combination of both methods. A distinct bacterial composition was revealed in the halitosis-associated biofilms compared with the health-associated biofilms. Actinomyces graevenitzii was shown to be significantly associated with the halitosis condition. The culture method identified 47 species, included Veillonella rogosae, never isolated from the tongue biofilm of halitosis patients so far. In the healthy condition, the culture-dependent method showed that the most frequent species were Streptococcus parasanguinis among the aerobes and Veillonella spp. among the anaerobes. The culture-independent cloning method detected more than 50 species. Streptococci, in particular S. mitis/oralis, S. pseudopneumoniae, and S. infantis as well as Prevotella spp., were found most frequently in halitosis patients. Streptococcus salivarius and Rothia mucilaginosa were found more frequently in the healthy condition.

CONCLUSIONS: The combination of the culture-dependent and culture-independent cloning techniques allowed for a widespread analysis of the tongue biofilm in halitosis patients. The results can support further pharmacological research for new antimicrobial agents and halitosis therapy strategies.

RevDate: 2019-11-14

Oppy CC, Jebeli L, Kuba M, et al (2019)

Loss of O-Linked Protein Glycosylation in Burkholderia cenocepacia Impairs Biofilm Formation and Siderophore Activity and Alters Transcriptional Regulators.

mSphere, 4(6): pii:4/6/e00660-19.

O-linked protein glycosylation is a conserved feature of the Burkholderia genus. The addition of the trisaccharide β-Gal-(1,3)-α-GalNAc-(1,3)-β-GalNAc to membrane exported proteins in Burkholderia cenocepacia is required for bacterial fitness and resistance to environmental stress. However, the underlying causes of the defects observed in the absence of glycosylation are unclear. Using proteomics, luciferase reporter assays, and DNA cross-linking, we demonstrate the loss of glycosylation leads to changes in transcriptional regulation of multiple proteins, including the repression of the master quorum CepR/I. These proteomic and transcriptional alterations lead to the abolition of biofilm formation and defects in siderophore activity. Surprisingly, the abundance of most of the known glycosylated proteins did not significantly change in the glycosylation-defective mutants, except for BCAL1086 and BCAL2974, which were found in reduced amounts, suggesting they could be degraded. However, the loss of these two proteins was not responsible for driving the proteomic alterations, biofilm formation, or siderophore activity. Together, our results show that loss of glycosylation in B. cenocepacia results in a global cell reprogramming via alteration of the transcriptional regulatory systems, which cannot be explained by the abundance changes in known B. cenocepacia glycoproteins.IMPORTANCE Protein glycosylation is increasingly recognized as a common posttranslational protein modification in bacterial species. Despite this commonality, our understanding of the role of most glycosylation systems in bacterial physiology and pathogenesis is incomplete. In this work, we investigated the effect of the disruption of O-linked glycosylation in the opportunistic pathogen Burkholderia cenocepacia using a combination of proteomic, molecular, and phenotypic assays. We find that in contrast to recent findings on the N-linked glycosylation systems of Campylobacter jejuni, O-linked glycosylation does not appear to play a role in proteome stabilization of most glycoproteins. Our results reveal that loss of glycosylation in B. cenocepacia strains leads to global proteome and transcriptional changes, including the repression of the quorum-sensing regulator cepR (BCAM1868) gene. These alterations lead to dramatic phenotypic changes in glycosylation-null strains, which are paralleled by both global proteomic and transcriptional alterations, which do not appear to directly result from the loss of glycosylation per se. This research unravels the pleiotropic effects of O-linked glycosylation in B. cenocepacia, demonstrating that its loss does not simply affect the stability of the glycoproteome, but also interferes with transcription and the broader proteome.

RevDate: 2019-11-13

Khan F, Nguyen Pham DT, Oloketuyi SF, et al (2019)

Antibiotics and their different application strategies in controlling the biofilm forming pathogenic bacteria.

Current pharmaceutical biotechnology pii:CPB-EPUB-102290 [Epub ahead of print].

BACKGROUND: The establishment of biofilm by most pathogenic bacteria has been known as one of the resistance mechanisms against antibiotics. Biofilm is a structural component where the bacterial community adheres to the biotic or abiotic surfaces by the help of extracellular polymeric substances (EPS) produced by bacterial cells. The biofilm matrix possesses the ability to resist several adverse environmental factors, including the effect of antibiotics. Therefore, the resistance of bacterial biofilm-forming cells could be increased up to 1000 times than the planktonic cells, hence requiring a significantly high concentration of antibiotics for treatment.

METHODS: Up to the present, several methodologies employing antibiotics as an anti-biofilm, anti-virulence or quorum quenching agent have been developed for the biofilm inhibition and eradication of pre-formed matured biofilm.

RESULTS: Among the anti-biofilm strategies being tested, the sub-minimal inhibitory concentration of several antibiotics either alone or in combination has been shown to inhibit the biofilm formation and down-regulate the production of virulence factors. The combinatorial strategies include (1) the combination of multiple antibiotics, (2) the combination of antibiotics with non-antibiotic agents and (3) loading of antibiotics onto a carrier.

CONCLUSION: The present review paper describes the role of several antibiotics as biofilm inhibitors and also the alternative strategies adopted for applications in eradicating and inhibiting the formation of biofilm by pathogenic bacteria.

RevDate: 2019-11-13

Van Dyck K, Van Dijck P, G Vande Velde (2020)

Bioluminescence Imaging to Study Mature Biofilm Formation by Candida spp. and Antifungal Activity In Vitro and In Vivo.

Methods in molecular biology (Clifton, N.J.), 2081:127-143.

The widespread use of indwelling medical devices has increased the number of device-related infections in hospitalized patients. These infections are often associated with the formation of biofilms on the medical implants that are difficult to treat because of their resistance to the classical antifungal drugs. The most common fungi isolated from catheters and other medical devices are Candida species. The Candida genus contains multiple species of which C. albicans and C. glabrata are the two most common pathogenic yeasts in humans. A limited number of animal models is available for investigating host-pathogen interactions and testing novel antifungal drugs in vivo against these species. Fungal load in biofilms in these models is traditionally analyzed postmortem, requiring host sacrifice and enumeration of microorganisms from individual biofilms in order to evaluate the amount of colony forming units and the efficacy of antifungal treatment. Bioluminescence imaging (BLI) made compatible with small animal models for in vivo biofilm formation is a valuable tool to follow biofilm development and its treatment longitudinally. Due to the noninvasive nature of BLI, the imaging procedure can be repeated in the same animal, allowing for follow-up of the biofilm growth in vivo without removing the implanted device or detaching the biofilm from its substrate. Although detecting a quantifiable in vivo BLI signal from biofilms formed on the inside of implanted catheters is challenging, BLI proved to be a practical tool in the study of fungal biofilms. This method describes the use of BLI for in vitro and in vivo follow-up of device-related fungal biofilm formation in mice and rats and antifungal activity testing against both C. albicans and C. glabrata device-associated biofilms. It can further be applied for efficient in vivo screening for interesting genes of the pathogen and the host involved in biofilm formation.

RevDate: 2019-11-13

Zhang Y, Zhao L, Song T, et al (2019)

Simultaneous nitrification and denitrification in an aerobic biofilm biosystem with loofah sponges as carriers for biodegrading hydrolyzed polyacrylamide-containing wastewater.

Bioprocess and biosystems engineering pii:10.1007/s00449-019-02247-x [Epub ahead of print].

Simultaneous nitrification and denitrification (SND) during treating hydrolyzed polyacrylamide (HPAM) containing wastewater were explored in an aerobic biofilm reactor biosystem. Here, loofah sponges as the environment-friendly and low-cost material were applied as the carriers in this biosystem. The removal efficiencies of HPAM and total nitrogen (TN) reached 43.6% and 54.3%, respectively, after 120 days stabilized running periods. Moreover, the structure of loofah sponges affected anaerobic microenvironment significantly which was indispensable for realizing a high-performance of SND. Key microorganisms in this biosystem included nitrobacteria, denitrobacteria and HPAM-biodegrading bacteria. The abundance of nitrobacteria and denitrobacteria on the biofilm was increased by 17.2% and 15.3%, respectively, through cultivation. Meanwhile, the biotransformation mechanisms of HPAM and diverse valence of nitrogen under different chemical oxygen demand (COD)/N and dissolved oxygen (DO) conditions were investigated. When COD/N and DO were 8:1 and 2 mg/L, HPAM biodegradation, SND efficiency and TN removal achieved their maximum, and the values were 54.3%, 92.3% and 60.1%, respectively. Key enzyme activities also reached their maximum in this condition. The optimal COD/N and DO was pivotal to achieve the high-performance of SND, and it was closely correlated with HPAM biodegradation. Meanwhile, SND could facilitate the biotransformation of HPAM.

RevDate: 2019-11-13

Ranjbar R, A Farahani (2019)

Study of genetic diversity, biofilm formation, and detection of Carbapenemase, MBL, ESBL, and tetracycline resistance genes in multidrug-resistant Acinetobacter baumannii isolated from burn wound infections in Iran.

Antimicrobial resistance and infection control, 8:172 pii:612.

Background: Antimicrobial resistance in multidrug-resistant Acinetobacter baumannii (MDR-AB) isolated from burn wound infections is a major concern in intensive care or burns units worldwide, and molecular studies are considered critical strategies for control of MDR-AB outbreaks in this regard. Thus, in this study, antibiotic resistance, biofilm-forming ability, molecular epidemiology of MDR A. baumannii strains recovered from patients with burns were investigated in three major hospital centers of Iran.

Methods: In this cross-sectional research, 163 non-repetitive A. baumannii strains were tested for susceptibility to antimicrobial agents. Polymerase chain reaction (PCR) was performed to characterize ambler classes A, B, and D β-lactamases, ISAba1 and integrons, biofilm formation was also investigated. Clonal relatedness was analyzed using Pulsed-Field Gel Electrophoresis (PFGE).

Results: Among 163 A. baumannii strains collected, 94.5% of them were Carbapenem-Non-Susceptible A. baumannii (CNSAB) and also 90.1 and 52.2% of them were Metallo-β-Lactamases (MBL) and Extended-Spectrum β-Lactamases (ESBL) producing isolates, respectively. Colistin and polymyxin B exhibited excellent activity against CNSAB strains. High prevalence of blaOXA - 23-like (85.1%), blaVIM (60.5%), blaPER - 1 (42.3%), tetB (67.8%), and Class 1 integrons (65.6%) were identified in CNSAB strains. ISAba1 element was associated with 42 (25.8%) and 129 (98.5%) of blaOXA-51-like and blaOXA-23-like genes, respectively. 6 clusters with the ability to form strong biofilms were found to be dominant and endemic in our entire areas.

Conclusions: Results of the present study show that antimicrobial resistance in CNSAB isolates from burn wound infections in monitored hospitals in Iran is multifactorial, and also findings of the study suggested that local antibiotic prescription policies should be regularly reviewed, and efficient infection control measures should be observed. Therefore, further strengthening of surveillance of antimicrobial resistance is urgently needed in these regions.

RevDate: 2019-11-13

Horváth B, Balázs VL, Varga A, et al (2019)

Preparation, characterisation and microbiological examination of Pickering nano-emulsions containing essential oils, and their effect on Streptococcus mutans biofilm treatment.

Scientific reports, 9(1):16611 pii:10.1038/s41598-019-52998-6.

Essential oils (EOs) are commonly applied in mouth care products like mouthwashes, mostly as an ethanolic solution or by usage of surfactants as solubilising agents. In this study, we present a formulation for preparation of Pickering nano-emulsions (PnE) of EOs as a novel form for application of EOs in mouth care. For the preparation of PnE, we have synthesised surface-modified silica nanoparticles with a mean diameter of 20 nm, as well as we have examined the effect of EOs concentration on PnE droplet size and stability. In vitro study of their effect on the Streptococcus mutans biofilm as the main pathogen of dental health problems has been performed. We have found that EOs in the PnE form has the highest effectiveness against biofilm formation. Diffusion through the biofilm model membrane was studied to explain this observation. We have found that PnEs have a better performance in the transportation of EOs trough model membrane than the ethanolic solutions and conventional emulsions (CEs).

RevDate: 2019-11-13

Vaishampayan A, E Grohmann (2019)

Multi-resistant biofilm-forming pathogens on the International Space Station.

Journal of biosciences, 44(5):.

The International Space Station (ISS) is a confined and closed habitat with unique conditions such as cosmic radiation, and microgravity. These conditions have a strong effect on the human and spacecraft microflora. They can affect the immune response of the crew-members, thus posing a threat to their health. Microbial diversity and abundance of microorganisms from surfaces, air filters and air samples on the ISS have been studied. Enterobacteriaceae, Bacillus spp., Propionibacterium spp., Corynebacterium spp., and Staphylococcus spp. were among the most frequently isolated bacteria. Microbial growth, biofilm formation, stress response, and pathogenicity are affected by microgravity. Increased resistance to antibiotics in bacteria isolated from the ISS has often been reported. Enterococcus faecalis and Staphylococcus spp. isolates from the ISS have been shown to harbor plasmid-encoded transfer genes. These genes facilitate the dissemination of antibiotic resistances. These features of ISS-pathogens call for novel approaches including highly effective antimicrobials which can be easily used on the ISS. A promising material is the antimicrobial surface coating AGXX, a self-recycling material consisting of two noble metals. It drastically reduced microbial growth of multi-resistant human pathogens, such as staphylococci and enterococci. Further novel approaches include the application of cold atmospheric plasma for the sterilization of spacecrafts.

RevDate: 2019-11-13

Manobala T, Shukla SK, Rao TS, et al (2019)

A new uranium bioremediation approach using radio-tolerant Deinococcus radiodurans biofilm.

Journal of biosciences, 44(5):.

Deinococcus radiodurans is the most radiation-tolerant organism ever known. It has gained importance in recent years as a potential candidate for bioremediation of heavy metals, especially the radioactive type. This study investigates the efficiency of a recombinant D. radiodurans (DR1-bf+) strain with an ability to form biofilm for uranium remediation. The modified Arsenazo III dye method was used to estimate the uranium concentration. Uranyl nitrate aqueous solution was generated during the operation of nuclear fuel reprocessing. The D. radiodurans biofilm (DR1-bf+) grown in the presence of 20 mM Ca2+ showed remarkable ability of uranyl ion removal. DR1-bf+ (Ca2+) biofilm removed ~75+/-2% of 1000 mg/L uranium within 30 min post-treatment from uranyl nitrate aqueous solution. Uranium removal rate was also found to be directly proportional to biofilm age. This study discusses the ability of D. radiodurans biofilm in uranium removal.

RevDate: 2019-11-13

Karley D, Shukla SK, TS Rao (2019)

Microbiota of spent nuclear fuel pool water with emphasis on their biofilm forming ability on stainless steel (SS-304L).

Journal of biosciences, 44(5):.

Spent nuclear fuel (SNF) pool is an essential unit of a nuclear power plant infrastructure, where radioactive fuel rods are kept for cooling and shielding, before reprocessing. This study explored the presence of bacteria in SNF pool water with emphasis on their capability to form biofilms on pool wall cladding material stainless steel (SS-304L). Bacteria were isolated from SNF pool water and were characterized using 16S rRNA gene sequencing. The six bacterial isolates (Bacillus subtilis, Staphylococcus sps., S. arlettae, S. epidermidis, S. auricularis and Chryseobacterium gleum) can grow and form biofilms at very low nutrient condition as well as in chronic radioactivity. The bacterial isolates formed biofilm on SS-304L and glass. However, the biofilm parameters assessed by CLSM microscopy showed that the strains preferred SS-304L surface for biofilm formation. On SS-304L, the maximum biomass (0.45 l μm3/l μm2) was formed by S. arlettae when compared to maximum biomass (0.054 l μm3/l μm2) by Staphylococcus sp., on glass. Maximum biofilm thickness on SS- 304L was observed by Staphylococcus sp. (8.81 l μm) when compared to that of S. epidermidis (4.16 l μm) on the glass surface. The biofilm formation on SS-304L surface suggests the possible risk of microbial-induced corrosion of SNF pool cladding material. This study highlights the need for mandatory monitoring of microbial biofilm formation in an extreme environment such as SNF pool.

RevDate: 2019-11-13

Padmavathi AR, Sriyutha Murthy P, Das A, et al (2019)

Copper oxide nanoparticles as an effective anti-biofilm agent against a copper tolerant marine bacterium, Staphylococcus lentus.

Biofouling [Epub ahead of print].

Biofilm formation on antifouling coatings is a serious concern in seawater cooling systems and the maritime industry. A prolific biofilm forming strain (Staphylococcus lentus), possessing high tolerance (>1,000 µg ml-1) to dissolved copper ions (Cu++) was isolated from titanium coupons exposed in the coastal waters of Kalpakkam, east coast of India. S. lentus formed increased biofilm (p < 0.05) at 100 µg ml-1 of Cu++ ions, when compared with the untreated control. To combat biofilm formation of this strain, the efficacy of copper oxide nanoparticles synthesized from copper nitrate by varying the concentrations of hexamine and cetyl trimethyl ammonium bromide (CTAB), was investigated. Complete (100%) inhibition of biofilm formation was observed with plain CuO NP (0.5 M hexamine, uncapped) at 1,000 µg ml-1. Capping with CTAB, influenced the morphology and the purity of the synthesized CuO NPs but did not alter their surface charge. Capping reduced metal ion release from CuO NPs and their antibacterial and anti-biofilm property against S. lentus. Overall, uncapped CuO NPs were effective in controlling biofilm formation of S. lentus. Concurrent release of copper ions and contact mediated physical damage by CuO NPs offer a promising approach to tackle metal tolerant biofilm bacteria.

RevDate: 2019-11-13

Najarzadeh Z, Mohammad-Beigi H, Nedergaard Pedersen J, et al (2019)

Plant Polyphenols Inhibit Functional Amyloid and Biofilm Formation in Pseudomonas Strains by Directing Monomers to Off-Pathway Oligomers.

Biomolecules, 9(11): pii:biom9110659.

Self-assembly of proteins to β-sheet rich amyloid fibrils is commonly observed in various neurodegenerative diseases. However, amyloid also occurs in the extracellular matrix of bacterial biofilm, which protects bacteria from environmental stress and antibiotics. Many Pseudomonas strains produce functional amyloid where the main component is the highly fibrillation-prone protein FapC. FapC fibrillation may be inhibited by small molecules such as plant polyphenols, which are already known to inhibit formation of pathogenic amyloid, but the mechanism and biological impact of inhibition is unclear. Here, we elucidate how polyphenols modify the self-assembly of functional amyloid, with particular focus on epigallocatechin gallate (EGCG), penta-O-galloyl-β-d-glucose (PGG), baicalein, oleuropein, and procyanidin B2. We find EGCG and PGG to be the best inhibitors. These compounds inhibit amyloid formation by redirecting the aggregation of FapC monomers into oligomeric species, which according to small-angle X-ray scattering (SAXS) measurements organize into core-shell complexes of short axis diameters 25-26 nm consisting of ~7 monomers. Using peptide arrays, we identify EGCG-binding sites in FapC's linker regions, C and N-terminal parts, and high amyloidogenic sequences located in the R2 and R3 repeats. We correlate our biophysical observations to biological impact by demonstrating that the extent of amyloid inhibition by the different inhibitors correlated with their ability to reduce biofilm, highlighting the potential of anti-amyloid polyphenols as therapeutic agents against biofilm infections.

RevDate: 2019-11-13

Janka E, Carvajal D, Wang S, et al (2019)

Treatment of Metformin-Containing Wastewater by a Hybrid Vertical Anaerobic Biofilm-Reactor (HyVAB).

International journal of environmental research and public health, 16(21): pii:ijerph16214125.

Several series of batch and continuous experiments were performed to investigate the removal of metformin and other contaminants from two wastewaters: wastewater I (WWI) containing 4 mg/L metformin and wastewater II (WWII) containing 110 g/L butanol. Biomethane potential (BMP) tests on WWII showed 77% of total chemical oxygen demand (tCOD = 110 g/L) degradability, and no apparent inhibition effects were observed. BMP tests on WWI showed an apparent inhibitory effect reflected in lower biogas production with increasing metformin concentration in the wastewater. Continuous flow hybrid vertical anaerobic biofilm (HyVAB®) experiments were consistent with the batch test findings. It was necessary to co-digest WWI (metformin) with WWII (easily degradable organics) to achieve complete metformin removal. After a period of adaptation, WWI and WWII co-digestion achieved up to 98% tCOD removal and 100% metformin removal. Most of the contaminants were removed in the anaerobic section of the HyVAB®, which implies that higher chemical oxygen demand (COD) loads than tested here are possible, given some optimization. The pilot reactor was able to manage organic loads of 11 g COD/d and above 10 mg/L metformin with a removal of 98% and 100% for tCOD and metformin, respectively.

RevDate: 2019-11-12

Huang Z, Wu L, Li X, et al (2019)

Zn(II) suppresses biofilm formation in Bacillus amyloliquefaciens by inactivation of the Mn(II) uptake.

Environmental microbiology [Epub ahead of print].

Biofilms are architecturally complex communities of microbial cells held together by a self-produced extracellular matrix. Considerable research has focused on the environmental signals that trigger or inhibit biofilm formation by affecting cellular signaling pathways; however, response to soil cues in plant-associated Bacillus has remained largely unaddressed. Therefore, we aimed to investigate the effect of Zn(II) ions in biofilm formation of Bacillus amyloliquefaciens FZB42. We demonstrated that biofilm formation of B. amyloliquefaciens FZB42 was abolished by Zn(II) at non-deleterious concentrations. Moreover, Zn(II) blocked matrix exopolysaccharide and TasA accumulations. Furthermore, the presence of Zn(II) suppressed expression of the response regulator Spo0F, but not of sensor histidine kinases KinA-D. Suppression of phosphorelay by excess Zn interferes with sinI induction under biofilm-inducing conditions, leading to repression of transcription of operons epsA-O and tapA-sigW-tasA. Addition of Zn(II) decreased the intracellular Mn(II) level by competing for binding to the solute-binding protein MntA during Mn(II) uptake. These results suggest that the metal ion Zn(II) has a negative effect on biofilm formation in the plant growth promoting and biocontrol bacterium B. amyloliquefaciens FZB42. This article is protected by copyright. All rights reserved.

RevDate: 2019-11-12

Peng J, Kumar K, Gross M, et al (2019)

Removal of Total Dissolved Solids (TDS) from Wastewater Using a Revolving Algal Biofilm Reactor.

Water environment research : a research publication of the Water Environment Federation [Epub ahead of print].

Total dissolved solids (TDS) comprising inorganic salts and organic matters are pollutants of concern to aquatic systems and water for human use. This work aimed to investigate the use of revolving algal biofilm (RAB) reactors as a sustainable and environmental friendly method to remove TDS from industrial effluents and municipal wastewaters. The wastewaters contained chloride, sodium, potassium, calcium, magnesium, and sulfate as the major components. The RAB reactors fed with synthetic industrial effluent with high TDS level demonstrated the best algal growth, with the highest TDS removal efficiency (27%) and removal rate (2,783 mg/L-day and 19,530 mg/m2 -day). A suspended algal culture system only removed 3% TDS from the same wastewater. The TDS removal by the RAB reactors was considered due to several mechanisms such as absorption by the algae cells, adsorption by extracellular polymeric substance (EPS) of the biofilm and/or precipitation. Collectively, this research shows that the RAB reactors can serve as an efficient system in wastewater remediation for TDS removal.

RevDate: 2019-11-12

Vaknin M, Steinberg D, Featherstone JD, et al (2019)

Exposure of Streptococcus mutans and Streptococcus sanguinis to blue light in an oral biofilm model.

Lasers in medical science pii:10.1007/s10103-019-02903-4 [Epub ahead of print].

The potential anti-cariogenic effect of blue light was evaluated using an oral biofilm model. Two species, Streptococcus mutans and Streptococcus sanguinis, were cultivated ex vivo on bovine enamel blocks for 24 h, either separately or mixed together, then exposed to blue light (wavelengths 400-500 nm) using 112 J/cm2. Twenty four or 48 h after exposure to light the biofilm structure and biomass were characterized and quantified using SEM and qPCR, respectively. Bacterial viability was analyzed by CLSM using live/dead bacterial staining. Gene expression was examined by RT-qPCR. After exposure to light, S. mutans biomass in mono-species biofilm was increased mainly by dead bacteria, relative to control. However, the bacterial biomass of S. mutans when grown in mixed biofilm and of S. sanguinis in mono-species biofilm was reduced after light exposure, with no significant change in viability when compared to control. Furthermore, when grown separately, an upregulation of gene expression related to biofilm formation of S. mutans, and downregulation of similar genes of S. sanguinis, were measured 24 h after exposure to blue light. However, in mixed biofilm, a downregulation of those genes in both species was observed, although not significant in S. mutans. In conclusion, blue light seems to effectively alter the bacterial biomass by reducing the viability and virulence characteristics in both bacterial species and may promote the anti-cariogenic balance between them, when grown in a mixed biofilm. Therefore, exposure of oral biofilm to blue light has the potential to serve as a complementary approach in preventive dentistry.

RevDate: 2019-11-12

Rollin-Pinheiro R, Rochetti VP, Dutra da Silva Xisto MI, et al (2019)

Sphingolipid biosynthetic pathway is crucial for growth, biofilm formation and membrane integrity of Scedosporium boydii.

Future medicinal chemistry [Epub ahead of print].

Aim: Glycosphingolipids are conserved lipids displaying a variety of functions in fungal cells, such as determination of cell polarity and virulence. They have been considered as potent targets for new antifungal drugs. The present work aimed to test two inhibitors, myriocin and DL-threo-1-Phenyl-2-palmitoylamino-3-morpholino-1-propanol, in Scedosporium boydii, a pathogenic fungus which causes a wide range of disease. Materials & methods: Mass spectrometry, microscopy and cell biology approaches showed that treatment with both inhibitors led to defects in fungal growth and membrane integrity, and caused an increased susceptibility to the current antifungal agents. Conclusion: These data demonstrate the antifungal potential of drugs inhibiting sphingolipid biosynthesis, as well as the usefulness of sphingolipids as promising targets for the development of new therapeutic options.

RevDate: 2019-11-12

Harro JM, Achermann Y, Freiberg JA, et al (2019)

Clearance of Staphylococcus aureus from In Vivo Models of Chronic Infection by Immunization Requires Both Planktonic and Biofilm Antigens.

Infection and immunity pii:IAI.00586-19 [Epub ahead of print].

Staphylococcus aureus is a causative agent of chronic biofilm-associated infections that are recalcitrant to resolution by the immune system or antibiotics. To combat these infections, an anti-staphylococcal, biofilm-specific quadrivalent vaccine against an osteomyelitis model in rabbits has previously been developed and shown to be effective at eliminating biofilm-embedded bacterial populations. However, the addition of antibiotics was required to eradicate remaining planktonic populations. In this study a planktonic up-regulated antigen was combined with the quadrivalent vaccine to remove the need for antibiotic therapy. Immunization with this pentavalent vaccine followed by intraperitoneal challenge of BALB/c mice with S. aureus resulted in 16.7% versus 91.7% mortality in pentavalent vaccine and control groups, respectively (p<0.001). Complete bacterial elimination was found in 66.7% of the pentavalent cohort, while only 8.3% of the control animals cleared the infection (p<0.05). Further protective efficacy was observed in immunized rabbits following intramedullary challenge with S. aureus, where 62.5% of the pentavalent cohort completely cleared the infection versus none of the control animals (p<0.05). Passive immunization of BALB/c mice with serum IgG against the vaccine antigens prior to intraperitoneal challenge with S. aureus prevented mortality in 100% of mice and eliminated bacteria in 33.3% of the challenged mice. These results demonstrate that targeting both the planktonic and biofilm stages with the pentavalent vaccine or the IgG elicited by immunization can effectively protect against S. aureus infection.

RevDate: 2019-11-11

Bridges AA, BL Bassler (2019)

The intragenus and interspecies quorum-sensing autoinducers exert distinct control over Vibrio cholerae biofilm formation and dispersal.

PLoS biology, 17(11):e3000429 pii:PBIOLOGY-D-19-02049 [Epub ahead of print].

Vibrio cholerae possesses multiple quorum-sensing (QS) systems that control virulence and biofilm formation among other traits. At low cell densities, when QS autoinducers are absent, V. cholerae forms biofilms. At high cell densities, when autoinducers have accumulated, biofilm formation is repressed, and dispersal occurs. Here, we focus on the roles of two well-characterized QS autoinducers that function in parallel. One autoinducer, called cholerae autoinducer-1 (CAI-1), is used to measure Vibrio abundance, and the other autoinducer, called autoinducer-2 (AI-2), is widely produced by different bacterial species and presumed to enable V. cholerae to assess the total bacterial cell density of the vicinal community. The two V. cholerae autoinducers funnel information into a shared signal relay pathway. This feature of the QS system architecture has made it difficult to understand how specific information can be extracted from each autoinducer, how the autoinducers might drive distinct output behaviors, and, in turn, how the bacteria use QS to distinguish kin from nonkin in bacterial communities. We develop a live-cell biofilm formation and dispersal assay that allows examination of the individual and combined roles of the two autoinducers in controlling V. cholerae behavior. We show that the QS system works as a coincidence detector in which both autoinducers must be present simultaneously for repression of biofilm formation to occur. Within that context, the CAI-1 QS pathway is activated when only a few V. cholerae cells are present, whereas the AI-2 pathway is activated only at much higher cell density. The consequence of this asymmetry is that exogenous sources of AI-2, but not CAI-1, contribute to satisfying the coincidence detector to repress biofilm formation and promote dispersal. We propose that V. cholerae uses CAI-1 to verify that some of its kin are present before committing to the high-cell-density QS mode, but it is, in fact, the broadly made autoinducer AI-2 that sets the pace of the V. cholerae QS program. This first report of unique roles for the different V. cholerae autoinducers suggests that detection of kin fosters a distinct outcome from detection of nonkin.

RevDate: 2019-11-11

Rossoni RD, de Barros PP, Lopes LADC, et al (2019)

Effects of surface pre-reacted glass-ionomer (S-PRG) eluate on Candida spp.: antifungal activity, anti-biofilm properties, and protective effects on Galleria mellonella against C. albicans infection.

Biofouling [Epub ahead of print].

Surface pre-reacted glass-ionomer (S-PRG) is a bioactive filler produced by PRG technology, which is applied to various dental materials. The inhibitory effects of S-PRG eluate against Candida, the most common fungal oral pathogen, were investigated. Minimum inhibitory concentrations (MIC) and anti-biofilm activities were tested against Candida albicans, Candida glabrata, Candida krusei, and Candida tropicalis. For the in vivo study, Galleria mellonella was used as a model to evaluate the effects of S-PRG on toxicity, hemocyte counts and candidiasis. The MIC of S-PRG ranged from 5 to 40% (v/v). S-PRG eluate exhibited anti-biofilm activity for all the Candida species tested. Furthermore, injection of S-PRG eluate into G. mellonella was not toxic to the larvae and protected G. mellonella against experimental candidiasis. In addition, S-PRG eluate inhibited biofilm formation by C. albicans, C. glabrata, C. krusei, and C. tropicalis and exerted protective effects on G. mellonella against experimental candidiasis in vivo.

RevDate: 2019-11-11

Lange MD, Farmer BD, J Abernathy (2019)

Vertebrate mucus stimulates biofilm development and upregulates iron acquisition genes in Flavobacterium columnare.

Journal of fish diseases [Epub ahead of print].

Columnaris disease is responsible for substantial losses throughout the production of many freshwater fish species. One of the ways in which the bacterium Flavobacterium columnare is so effective in initiating disease is through the formation of biofilms on fish skin and gills. To further explore the interaction between host factors and bacterial cells, we assayed the ability of vertebrate mucus to enhance F. columnare biofilm development. Different concentrations of catfish, tilapia and pig mucus (5-60 µg/ml) increased biofilm growth at varying degrees among F. columnare isolates. Our data suggest that vertebrate mucus acts as a signalling molecule for the development of F. columnare biofilms; however, there are clear disparities in how individual isolates respond to different mucus fractions to stimulate biofilms. The expression of iron acquisition genes among two genomovar II isolates showed that ferroxidase, TonB receptor and the siderophore synthetase gene were all significantly upregulated among F. columnare biofilms. Interestingly, the siderophore acetyltransferase gene was only shown to be significantly upregulated in one of the genomovar II isolates. This work provides insight into our understanding of the interaction between F. columnare and vertebrate mucus, which likely contributes to the growth of planktonic cells and the transition into biofilms.

RevDate: 2019-11-11

Wang Z, Chen XM, Ni BJ, et al (2019)

Model-based assessment of chromate reduction and nitrate effect in a methane-based membrane biofilm reactor.

Water research X, 5:100037 pii:100037.

Chromate contamination can pose a high risk to both the environment and public health. Previous studies have shown that CH4-based membrane biofilm reactor (MBfR) is a promising method for chromate removal. In this study, we developed a multispecies biofilm model to study chromate reduction and its interaction with nitrate reduction in a CH4-based MBfR. The model-simulated results were consistent with the experimental data reported in the literature. The model showed that the presence of nitrate in the influent promoted the growth of heterotrophs, while suppressing methanotrophs and chromate reducers. Moreover, it indicated that a biofilm thickness of 150 μm and an influent dissolved oxygen concentration of 0.5 mg O2/L could improve the reactor performance by increasing the chromate removal efficiency under the simulated conditions.

RevDate: 2019-11-11

She P, Zhou L, Li S, et al (2019)

Synergistic Microbicidal Effect of Auranofin and Antibiotics Against Planktonic and Biofilm-Encased S. aureus and E. faecalis.

Frontiers in microbiology, 10:2453.

Methicillin-resistant/susceptible Staphylococcus aureus (MRSA/MSSA) and Enterococcus faecalis strains are often found in community- and hospital-acquired infections. The single use of conventional antibiotics hardly completely kills the bacterial cells of interest, especially in the form of biofilms. Thus, drug repurposing and antimicrobial combination are promising ways to solve this problem. Antimicrobial susceptibility assays against cocci in a suspension and in a biofilm mode of growth were performed with broth microdilution methods. Checkerboard assays and the cutaneous mouse infection model were used to examine the activity of auranofin and conventional antibiotics alone and in combination. In the present study, auranofin possesses potent antimicrobial activities against both planktonic cells and biofilms with minimum inhibitory concentrations ranging 0.125-0.5 mg/L. Auranofin in combination with linezolid or fosfomycin showed synergistic antimicrobial activities against S. aureus MSSA and MRSA both in vitro and in vivo. Similarly, auranofin also behaved synergistic effect with chloramphenicol against E. faecalis. Additionally, auranofin improved the antibiofilm efficacy of chloramphenicol and linezolid, even on the biofilms grown on a catheter surface. Though, S. epidermidis showed significant susceptibility to AF treatment, no synergistic antimicrobial effects were observed with antibiotics we tested. In all, the use of a combination of auranofin with linezolid, fosfomycin, and chloramphenicol can provide a synergistic microbicidal effect in vitro and in vivo, which rapidly enhances antimicrobial activity and may help prevent or delay the emergence of resistance.

RevDate: 2019-11-11

Millen S, Gross C, Donhauser N, et al (2019)

Collagen IV (COL4A1, COL4A2), a Component of the Viral Biofilm, Is Induced by the HTLV-1 Oncoprotein Tax and Impacts Virus Transmission.

Frontiers in microbiology, 10:2439.

Human T-cell leukemia virus type 1 (HTLV-1) is the etiologic agent for Adult T-Cell Leukemia/Lymphoma (ATLL) and HTLV-1-Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP). HTLV-1 infects CD4+ T-cells via cell-to-cell transmission requiring reorganization of the cytoskeleton and expression of the viral transactivator and oncoprotein Tax. Viruses spread at the virological synapse (VS), a virus-induced specialized cell-cell contact, by polarized budding into synaptic clefts, and by cell surface transfer of viral biofilms (VBs). Since little is known about Tax's role in formation of the VB, we asked which component of the VB is regulated by Tax and important for HTLV-1 transmission. Collagens are not only structural proteins of the extracellular matrix and basal membrane but also represent an important component of the VB. Here, we report that among the collagens known to be present in VBs, COL4 is specifically upregulated in the presence of HTLV-1 infection. Further, we found that transient expression of Tax is sufficient to induce COL4A1 and COL4A2 transcripts in Jurkat and CCRF-CEM T-cells, while robust induction of COL4 protein requires continuous Tax expression as shown in Tax-transformed T-cell lines. Repression of Tax led to a significant reduction of COL4A1/A2 transcripts and COL4 protein. Mechanistically, luciferase-based promoter studies indicate that Tax activates the COL4A2 and, to a less extent, the COL4A1 promoter. Imaging showing partial co-localization of COL4 with the viral Gag protein in VBs at the VS and transfer of COL4 and Gag to target cells suggests a role of COL4 in VB formation. Strikingly, in chronically infected C91-PL cells, knockout of COL4A2 impaired Gag transfer between infected T-cells and acceptor T-cells, while release of virus-like particles was unaffected. Taken together, we identified COL4 (COL4A1, COL4A2) as a component of the VB and a novel cellular target of Tax with COL4A2 appearing to impact virus transmission. Thus, this study is the first to provide a link between Tax's activity and VB formation by hijacking COL4 protein functions.

RevDate: 2019-11-10

Das P, VS Karankar (2019)

New avenues of controlling microbial infections through anti-microbial and anti-biofilm potentials of green mono-and multi-metallic nanoparticles: A review.

Journal of microbiological methods pii:S0167-7012(19)30426-9 [Epub ahead of print].

Nanoparticles synthesized through the green route deserve special mention because this green technology is not only energy-efficient and cost-effective but also amenable to the environment. Various biological resources have been used for the generation of these 'green nanoparticles'. Biological wastes have also been focused in this direction thereby promoting the value of waste. Reports indicate that green nanoparticles exhibit remarkable antimicrobial activities, both singly as well as in combination with standard antibiotics. The current phenomenon of multi-drug resistance has resulted due to indiscriminate administration of high-doses of antibiotics followed by significant toxicity. In the face of this emergence of drug-resistant microbes, the efficacy of green nanoparticles might prove greatly beneficial. Microbial biofilm is another hurdle in the effective treatment of diseases as the microorganisms, being embedded in the meshwork of the biofilm, evade the antimicrobial agents. Nanoparticles may act as a ray of hope on the face of this challenge too, as they not only destroy the biofilms but also lessen the doses of antibiotics required, when administered in combination with the nanoparticles. It should be further noted that the resistance mechanisms exhibited by the microorganisms seem not that relevant for nanoparticles. The current review, to the best of our knowledge, focuses on the structures of these green nanoparticles along with their biomedical potentials. It is interesting to note how a variety of structures are generated by using resources like microbes or plants or plant products and how the structure affects their activities. This study might pave the way for further development in this arena and future work may be taken up in identifying the detailed mechanism by which 'green' synthesis empowers nanoparticles to kill pathogenic microbes.

RevDate: 2019-11-10

Lawrence JR, Paule A, Swerhone GDW, et al (2019)

Microscale and molecular analyses of river biofilm communities treated with microgram levels of cerium oxide nanoparticles indicate limited but significant effects.

Environmental pollution (Barking, Essex : 1987) pii:S0269-7491(19)32179-7 [Epub ahead of print].

Cerium oxide (CeO2) nanoparticles are used as in-fuel catalysts and in manufacturing processes, creating a potential for release to aquatic environments. Exposures at 1 and 10 μg/L CeO2-nanoparticles were made to assess effects during the development of river biofilm communities. Scanning transmission x-ray microscopy (STXM) indicated extensive sorption of nanoparticles to the community and co-localization with lipid moieties. Following 8 weeks of development, polycarbonate coupons were removed from the reactors and used for molecular analyses, denaturing gradient gel electrophoresis analysis (DGGE-16S rRNA) and 16S rRNA amplicon sequencing. Microscopic imaging of the biofilm communities (bacterial, photosynthetic biomass, exopolymer composition, thickness, protozoan numbers), as well as carbon substrate utilization fingerprinting was performed. There was a trend toward reduced photosynthetic biomass, but no significant effects of CeO2 exposure were found on photosynthetic and bacterial biomass or biofilm thickness. Sole carbon source utilization analyses indicated increased utilization of 10 carbon sources in the carbohydrate, carboxylic acid and amino acids categories related to CeO2 exposures; however, predominantly, no significant effects (p < 0.05) were detected. Measures of microbial diversity, lectin binding affinities of exopolymeric substances and results of DGGE analyses, indicated significant changes to community composition (p < 0.05) with CeO2 exposure. Increased binding of the lectin Canavalia ensiformis was observed, consistent with changes in bacterial-associated polymers. Whereas, no significant changes were observed in binding to residues associated with algal and cyanobacterial exopolymers. 16S rRNA amplicon sequencing of community DNA indicated changes in diversity and shifts in community composition; however, these did not trend with increasing CeO2 exposure. Counting of protozoans in the biofilm communities indicated no significant effects on this trophic level. Thus, based on biomass and functional measures, CeO2 nanoparticles did not appear to have significant effects; however, there was evidence of selection pressure resulting in significant changes in microbial community composition.

RevDate: 2019-11-09

Keren-Paz A, I Kolodkin-Gal (2019)

A brick in the wall: Discovering a novel mineral component of the biofilm extracellular matrix.

New biotechnology pii:S1871-6784(18)31823-5 [Epub ahead of print].

Multicellular bacterial communities, known as biofilms, have been thought to be held together solely by a self-produced organic extracellular matrix (ECM). However, new evidence for a missed mineral constituent of ECM in both Gram-positive and Gram-negative bacterial species, is accumulating. Study of two phylogenetically distinct bacteria, Bacillus subtilis and Mycobacterium smegmatis, identified a novel mechanism crucial for proper biofilm development and architecture - an active, genetically regulated, production of crystalline calcite. The calcite scaffolds stabilize bacterial biofilms, limit penetration of small molecule solutes such as antibiotics and play a conserved role in the assembly of those complex differentiated multicellular communities. Accumulating evidence suggests tight biological regulation of the formation of these functional minerals. This review discusses the recently discovered structural and functional roles of extracellular minerals in biofilms. It is proposed that it is time for a more complete view of the ECM as a complex combination of organic and nonorganic materials, especially in the light of the possible implications for treatment of biofilm infections.

RevDate: 2019-11-09

Sood U, Singh DN, Hira P, et al (2019)

Rapid and solitary production of mono-rhamnolipid biosurfactant and biofilm inhibiting pyocyanin by a taxonomic outlier Pseudomonas aeruginosa strain CR1.

Journal of biotechnology pii:S0168-1656(19)30915-0 [Epub ahead of print].

Biosurfactant - Rhamnolipids (RLs) and antibacterial toxin - pyocyanin (PYO) produced by Pseudomonas aeruginosa strains have great potential for biotechnological applications. Generally, RLs are produced as a mixture of di-rhamnolipids (di-RLs) and mono-rhamnolipids (mono-RLs). Mono-RLs possess superior emulsification and antimicrobial properties and are costlier than di-RLs. In this study, a taxonomic outlier P. aeruginosa strain CR1 isolated from rhizosphere soil was explored for mono-RLs and PYO production. Phylogenetically strain CR1 resembles avirulent outlier P. aeruginosa strain ATCC9027, lacks archetypical virulence genes and harbors unique pathways for the synthesis of solely mono-RLs and PYO. Strain CR1 produced RL biosurfactant which efficiently emulsified hydrocarbons, showed hemolysis and inhibited Bacillus subtilis. At 37 °C, strain CR1 exclusively produced 21.77 g L-1 and 19.22 g L-1 rhamnolipid in glycerol amended Luria Bertani (LB) medium and basal medium amended with rice bran oil, respectively after 54 h growth. Besides RL production was unaffected under varying nitrogen sources. Structural characterization using FTIR, TLC, and LC-MS confirmed that strain CR1 exclusively produced mono-RLs, majorly dominated by Rha-C10-C10, Rha-C10-C8, and CH3-Rha-C12:2-C10:1. The compound was stable over a wide pH range (4-12), salinity (25%) and 100 °C indicating its applicability under harsh environmental conditions. In addition, strain CR1 produced 4.5 µg mL-1 PYO, which could efficiently inhibit biofilm formation by Bacillus species. The environmental outlier strain CR1 can be used for the industrial production of biotechnologically important mono-RLs and PYO.

RevDate: 2019-11-09

Marquès C, Collin V, Franceschi C, et al (2019)

Fosfomycin and Staphylococcus aureus: transcriptomic approach to assess effect on biofilm, and fate of unattached cells.

The Journal of antibiotics pii:10.1038/s41429-019-0256-y [Epub ahead of print].

Interest has been rekindled in the old antibiotic fosfomycin, partly because of its ability to penetrate biofilm. Using a transcriptomic approach, we investigated the modifications induced by fosfomycin in sessile cells of a clinical Staphylococcus aureus isolated from a device-associated infection. Cells still able to form biofilm after 4 h of incubation in the presence of subinhibitory concentrations of fosfomycin and cells from 24-h-old biofilm later submitted to fosfomycin had 6.77% and 9.41%, respectively, of differentially expressed genes compared with their antibiotic-free control. Fosfomycin induced mostly downregulation of genes assigned to nucleotide, amino acid and carbohydrate transport, and metabolism. Adhesins and capsular biosynthesis proteins encoding genes were downregulated in fosfomycin-grown biofilm, whereas the murein hydrolase regulator lgrA and a D-lactate dehydrogenase-encoding gene were upregulated. In fosfomycin-treated biofilm, the expression of genes encoding adhesins, the cell wall biosynthesis protein ScdA, and to a lesser extent the fosfomycin target MurA was also decreased. Unattached cells surrounding fosfomycin-grown biofilm showed greater ability to form aggregates than their counterparts obtained without fosfomycin. Reducing their global metabolism and lowering cell wall turnover would allow some S. aureus cells to grow in biofilm despite fosfomycin stress while promoting hyperadherent phenotype in the vicinity of the fosfomycin-treated biofilm.

RevDate: 2019-11-09

Rehman ZU, Fortunato L, Cheng T, et al (2019)

Metagenomic analysis of sludge and early-stage biofilm communities of a submerged membrane bioreactor.

The Science of the total environment, 701:134682 pii:S0048-9697(19)34673-X [Epub ahead of print].

Biofilm formation on membranes in activated sludge membrane bioreactors (MBR), commonly identified as biofouling, is a significant problem for MBR operations. A better understanding of microbial species involved in the biofilm formation is needed to develop anti-biofilm measures. A read-based and genome-resolved shotgun metagenomic approach was applied to characterize the composition and functional potential of the sludge and early stage biofilm microbial communities in an MBR process. Read-based analysis revealed that the prevalence of different phyla are relatively similar in both the sludge and biofilm samples, with Proteobacteria as the most dominant, followed by Chloroflexi, Bacteroidetes and Planctomycetes. However, the relative abundance of these phyla slightly varies between the sludge and biofilm. Phyla such as Actinobacteria, bacterial candidate phyla, Chlamydiae, Cyanobacteria/Melainabacteria and Firmicutes are 2 to 4 times more abundant in the biofilm than in the sludge. At the genus level, genera belonging to Proteobacteria (Legionella, Caulobacter, Sphingomonas, Acinetobacter and Rhizobium), Cyanobacteria (Hassallia), and Spirochaetes (Turneriella) are at least twice more abundant in the biofilm. These genera, especially those belonging to Phylum Proteobacteria, are known to play an important role in the formation of biofilms on surfaces. The Alpha diversity is found slightly higher in the biofilm, compared with sludge samples. Functional classification of reads through the SEED subsystem shows that functional classes such as those involved in the metabolism of various molecules are significantly different in the biofilm and sludge. A phylogenomic analysis of the six extracted metagenome assembled genomes (MAGs) shows that three MAGs belong to Proteobacteria, and one MAG belong to each of Chloroflexi, Bacteroidetes and Planctomycetes. The relative abundance of the MAG belonging to Alphaproteobacteria is higher in the biofilm. A functional potential analysis of the MAGs reveals their potential to metabolize carbon and nitrogen sources, as well as the prevalence of antibiotic resistance genes.

RevDate: 2019-11-09

Alfaifi AA, Lin WS, Aldhaian BA, et al (2019)

Impact of caffeine on metabolic activity and biofilm formation of Candida albicans on acrylic denture resin in the presence of nicotine.

The Journal of prosthetic dentistry pii:S0022-3913(19)30599-2 [Epub ahead of print].

STATEMENT OF PROBLEM: Candida albicans has been implicated in denture stomatitis, and this effect is exacerbated by nicotine exposure. However, studies have also suggested that caffeine exposure inhibits the growth of C. albicans. The interaction effects of nicotine and caffeine are not yet clear on the growth of C. albicans.

PURPOSE: The purpose of this in vitro study was to determine the effect of caffeine on metabolic activity and biofilm formation of C. albicans growing on acrylic denture resin while simultaneously exposed to nicotine and, if an effect were to be identified, whether this effect would vary depending on the caffeine concentration.

MATERIAL AND METHODS: A total of 240 acrylic resin specimens were divided into 2 equal groups (120 each). Specimens in one group were processed to measure C. albicans metabolic activity, and those in the other group were processed to measure C. albicans biofilm attachment. Ten subgroups (n=12) were established within each group with different concentration combinations of nicotine and caffeine to test the interaction effect. The first subgroup was designed as a negative control, containing 0 mg/mL of nicotine and caffeine. The following subgroups all contained 8.00 mg/mL of nicotine, and the caffeine concentrations were prepared at the following 9 levels: 0, 0.25, 0.50, 1.00, 2.00, 4.00, 8.00, 16.00, and 32.00 mg/mL. Metabolic activity was measured by using a 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-carboxanilide (XTT) assay. Biofilm attachment was measured by using spiral plating and calculated in terms of the number of colony-forming units (CFUs)/mL. Descriptive statistics and a 2-way ANOVA were conducted to determine whether the concentrations of nicotine and caffeine used affected the biofilm attachment and metabolic activity of C. albicans (α=.05).

RESULTS: The presence of 8 mg/mL of nicotine increased the metabolic activity and biofilm formation of C. albicans. When compared with the 0 mg/mL of caffeine and 8.00 mg/mL of nicotine group, caffeine from 1.00 to 4.00 mg/mL significantly increased C. albicans biofilm metabolic activity. Caffeine at 16.00 and 32.00 mg/mL significantly decreased C. albicans biofilm metabolic activity in the presence of 8 mg/mL of nicotine. Caffeine from 1.00 to 32.00 mg/mL significantly decreased the biofilm formation of C. albicans in the presence of 8 mg/mL of nicotine.

CONCLUSIONS: The presence of 8 mg/mL of nicotine alone increased the metabolic activity and biofilm formation of C. albicans. In the presence of 8 mg/mL of nicotine with different caffeine concentrations, the results suggest that, overall, caffeine at higher concentrations (16 and 32 mg/mL) inhibited the metabolic activity and biofilm formation of C. albicans on acrylic denture resin most.

RevDate: 2019-11-09

Liu F, Sun Z, Wang F, et al (2020)

Inhibition of biofilm formation and exopolysaccharide synthesis of Enterococcus faecalis by phenyllactic acid.

Food microbiology, 86:103344.

This study aimed to evaluate the inhibitory activity of phenyllactic acid (PLA) against the biofilm formation of Enterococcus faecalis and to explore its potential molecular mechanism. The MIC value of PLA that inhibited the growth of E. faecalis R612-Z1 in BHI broth was 5 mg/mL. PLAs at subinhibitory concentrations of 1.25 and 2.50 mg/mL were found to inhibit biofilm formation by a crystal violet staining assay. The cell swimming and swarming motilities of E. faecalis were reduced in the presence of PLA. An apparent decrease in the thickness of PLA-treated biofilms was observed through confocal laser scanning microscopy analysis. The exopolysaccharide production in E. faecalis biofilms was inhibited by EPS quantification assay and scanning electron microscopy (SEM). qRT-PCR analyses showed that PLA down-regulated the transcription of Ebp pili genes (ebpABC) and Epa polysaccharide genes (epaABE). PLA inhibited the biofilm formation by interfering with cell mobility and EPS production of E. faecalis. In addition, PLA at concentrations of 10.0 mg/mL can effectively control the bacterial cells in a three-day-old mature biofilm of E. faecalis grown on 24-well flat-bottom polystyrene plates and stainless-steel surfaces. Thus, PLA is potentially an effective agent to control E. faecalis biofilms.

RevDate: 2019-11-09

Guo D, Wang S, Li J, et al (2020)

The antimicrobial activity of coenzyme Q0 against planktonic and biofilm forms of Cronobacter sakazakii.

Food microbiology, 86:103337.

Coenzyme Q0 (CoQ0) has demonstrated antitumor, anti-inflammatory, and anti-angiogenic activities. Cronobacter sakazakii is an opportunistic foodborne pathogen associated with high mortality in neonates. In this study, the antimicrobial activity and possible antimicrobial mechanism of CoQ0 against C. sakazakii were investigated. Moreover, the inactivation effect of CoQ0 on C. sakazakii in biofilms was also evaluated. The minimum inhibitory concentration (MIC) of CoQ0 against C. sakazakii strains ranged from 0.1 to 0.2 mg/mL. Treatment caused cell membrane dysfunction, as evidenced by cell membrane hyperpolarization, decreased intracellular ATP concentration and cell membrane integrity, and changes in cellular morphology. CoQ0 combined with mild heat treatment (45, 50, or 55 °C) decreased the number of viable non-desiccated and desiccated C. sakazakii cells in a time- and dose-dependent manner in reconstituted infant milk. Furthermore, CoQ0 showed effective inactivation activity against C. sakazakii in biofilms on stainless steel, reducing the number of viable cells and damaging the structure of the biofilm. These findings suggest that CoQ0 has a strong inactivate effect on C. sakazakii and could be used in food production environments to effectively control C. sakazakii and reduce the number of illnesses associated with it.

RevDate: 2019-11-09

Li T, Sun X, Chen H, et al (2020)

Methyl anthranilate: A novel quorum sensing inhibitor and anti-biofilm agent against Aeromonas sobria.

Food microbiology, 86:103356.

Quorum sensing (QS), bacterial cell-to-cell communication, is a gene regulatory mechanism that regulates virulence potential and biofilm formation in many pathogens. Aeromonas sobria, a common aquaculture pathogen, was isolated and identified by our laboratory from the deteriorated turbot, and its potential for virulence factors and biofilm production was regulated by QS system. In view of the interference with QS system, this study was aimed to investigate the effect of methyl anthranilate at sub-Minimum Inhibitory Concentrations (sub-MICs) on QS-regulated phenotypes in A. sobria. The results suggested that 0.5 μL/mL of methyl anthranilate evidently reduced biofilm formation (51.44%), swinging motility (74.86%), swarming motility (71.63%), protease activity (43.08%), and acyl-homoserine lactone (AHL) production. Furthermore, the real-time quantitative PCR (RT-qPCR) and in silico analysis showed that methyl anthranilate might inhibit QS system in A. sobria by interfering with the biosynthesis of AHL, as well as competitively binding with receptor protein. Therefore, our data indicated the feasibility of methyl anthranilate as a promising QS inhibitor and anti-biofilm agent for improving food safety.

RevDate: 2019-11-08

Harrison A, Hardison RL, Wallace RM, et al (2019)

Reprioritization of biofilm metabolism is associated with nutrient adaptation and long-term survival of Haemophilus influenzae.

NPJ biofilms and microbiomes, 5:33 pii:105.

Nontypeable Haemophilus influenzae (NTHI) is a human-restricted pathogen with an essential requirement for heme-iron acquisition. We previously demonstrated that microevolution of NTHI promotes stationary phase survival in response to transient heme-iron restriction. In this study, we examine the metabolic contributions to biofilm formation using this evolved NTHI strain, RM33. Quantitative analyses identified 29 proteins, 55 transcripts, and 31 metabolites that significantly changed within in vitro biofilms formed by RM33. The synthesis of all enzymes within the tryptophan and glycogen pathways was significantly increased in biofilms formed by RM33 compared with the parental strain. In addition, increases were observed in metabolite transport, adhesin production, and DNA metabolism. Furthermore, we observed pyruvate as a pivotal point in the metabolic pathways associated with changes in cAMP phosphodiesterase activity during biofilm formation. Taken together, changes in central metabolism combined with increased stores of nutrients may serve to counterbalance nutrient sequestration.

RevDate: 2019-11-08

Teves A, Blanco D, Casaretto M, et al (2019)

Effectiveness of different disinfection techniques of the root canal in the elimination of a multi-species biofilm.

Journal of clinical and experimental dentistry, 11(11):e978-e983 pii:56000.

Background: The purpose of the study was to evaluate the effectiveness of different root canal disinfection techniques in the elimination of a multi-species biofilm from inside the root canal.

Material and Methods: Fifty mandibular first premolars were used in the present study, standardized to 11mm of root length, and instrumented with a reciprocation system Reciproc, (VDW GmbH, Munich, Germany) to a #50. Longitudinally sectioned halves of the roots were obtained and washed with NaOCl 4%, EDTA 17% and 5% sodium thiosulfate, and sterilized by autoclaving for 15 minutes at 121°C. A multi-species biofilm broth was developed with three strains of bacteria under laboratory conditions: Enterococcus faecalis ATTC 29212, Eikenella corrodens ATTC 23834, Streptococcus anginosus ATTC 33397. Roots were autoclaved and transferred to the broth for 4 days and then were subjected to either disinfection with sodium hypochlorite 4% and XP-endo Finisher (FKG Dentaire, La Chaux-de-Fonds, Switzerland) or chlorhexidine 2% with and without activation with XP-endo Finisher (FKG Dentaire, La Chaux-de-Fonds, Switzerland).

Results: The evaluations of the biofilm elimination showed results that indicate that the 4% sodium hypochlorite group with positive pressure irrigation presented significant differences with the group that had irrigation with sodium hypochlorite activated with XP-endo Finisher and the chlorhexidine groups to 2% (P<0.05).

Conclusions: Chlorhexidine 2% activated with the XP-endo Finisher does not exert elimination or improved cleaning effect on the multi-species biofilm. Activation of sodium hypochlorite 4% improved the elimination of the multi-species biofilm. Key words:Biofilm, multispecies, chlorhexidine, sodium hypochlorite.

RevDate: 2019-11-08

Theodora NA, Dominika V, DE Waturangi (2019)

Screening and quantification of anti-quorum sensing and antibiofilm activities of phyllosphere bacteria against biofilm forming bacteria.

BMC research notes, 12(1):732 pii:10.1186/s13104-019-4775-1.

OBJECTIVE: The objectives of this research were to screen anti-quorum sensing activity of phyllosphere bacteria and quantify their antibiofilm activity against biofilm forming bacteria (Bacillus cereus, Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium, Vibrio cholerae, Pseudomonas aeruginosa).

RESULTS: We found 11 phyllosphere bacteria isolates with potential anti-quorum sensing activity. Most of the crude extracts from phyllosphere bacteria isolates had anti-quorum sensing activity against Chromobacterium violaceum at certain concentration (20 and 10 mg/mL), but not crude extract from isolate JB 7F. Crude extract showed the largest turbid zone (1,27 cm) using isolate JB 14B with concentration of 10 mg/mL and the narrowest turbid zone isolate (1 cm) using JB 18B with concentration of 10 mg/mL. Crude extracts showed various antibiofilm activities against all tested pathogenic bacteria, it showed the highest biofilm inhibition (90%) and destruction activities (76%) against S. aureus.

RevDate: 2019-11-07

Nagaraju VT, Ray AK, Avunje S, et al (2019)

Vibrio harveyi biofilm as immunostimulant candidate for high-health pacific white shrimp, Penaeus vannamei farming.

Fish & shellfish immunology pii:S1050-4648(19)31047-2 [Epub ahead of print].

The study was to develop Vibrio harveyi biofilm-based novel microbial product and its oral delivery for high health Penaeus vannamei farming. Yield of bacterial biofilm was optimized on chitin substrate (size: <360, 360-850 and 850-1250 μm; concentration: 0.3, 0.6 and 0.9%) in tryptone soy broth (0.15%). The biofilm was characterized by crystal violet assay, SEM and LSCM imaging; protein profiling by SDS-PAGE and LC-ESI-MS/MS. The immune stimulatory effect of the biofilm in yard experiments was evaluated by relative quantification of immune genes using real-time PCR effect on overall improvement on health status under field trials. The highest biofilm yield (6.13 ± 0.2 × 107 cfu/ml) was obtained at 0.6% of <360 μm chitin substrate. The biofilm formation was stabilized by 96 h of incubation at 30 °C. Protein profiling confirmed expression of six additional proteins (SDS-PAGE) and 11 proteins were differentially expressed (LC-ESI-MS/MS) in biofilm cells over free cells of V. harveyi. Oral administration of the biofilm for 48 h confirmed to enhance expression of antimicrobial peptides, penaeidin, crustin and lysozyme in P. vannamei. Further Oral administration of biofilm for two weeks to P. vannamei (1.8 ± 0.13 g) improved the growth (2.66 ± 0.06 g) and survival (84.44 ± 1.82%) compared to control (2.15 ± 0.03 g; 70.94 ± 0.66%) Nursery trials showed a significant reduction in occurrence of anatomical deformities like antenna cut (12.67 ± 0.66%), rostrum cut (4.66 ± 0.87%), and tail rot (3.33 ± 0.88%), compared to animals fed with normal diet which was 24.33 ± 2.72; 14 ± 1.52 and 10.66 ± 1.45% respectively. In vitro and in vivo studies suggest inactivated biofilm cells of V. harveyi on chitin substrate express additional antigenic proteins and when administered orally through feed at regular intervals stimulates immune response and improve growth, survival and health status of shrimp.

RevDate: 2019-11-07

Brunetti G, Navazio AS, Giuliani A, et al (2019)

Candida blood stream infections observed between 2011 and 2016 in a large Italian University Hospital: A time-based retrospective analysis on epidemiology, biofilm production, antifungal agents consumption and drug-susceptibility.

PloS one, 14(11):e0224678 pii:PONE-D-19-18698.

Candida bloodstream infection (BSI) represents a growing infective problem frequently associated to biofilm production due to the utilization of intravascular devices. Candida species distribution (n = 612 strains), their biofilm production and hospital antifungal drug consumption were evaluated in different wards of a tertiary care academic hospital in Italy during the years 2011-2016. In the considered time window, an increasing number of Candida BSI (p = 0.005) and of biofilm producing strains were observed (p<0.0001). Although C. albicans was the species more frequently isolated in BSI with a major biofilm production, an increased involvement of non-albicans species was reported, particularly of C. parapsilosis that displayed a high frequency in catheter infections, and lower biofilm production compared to C. albicans. Although trends of biofilm production were substantially stable in time, a decreasing biofilm production by C. parapsilosis in the Intensive Care Unit (ICU) was observed (p = 0.0041). Principal component analysis displayed a change in antifungal drugs consumption driven by two mutually independent temporal trends, i.e. voriconazole use in the general medicine wards initially, and fluconazole use mainly in the ICU; these factors explain 68.9% and 25.7% of total variance respectively. Moreover, a significant trend (p = 0.003) in fluconazole use during the whole time period considered emerged, particularly in the ICU (p = 0.017), but also in the general medicine wards (p = 0.03). These trends paralleled with significant increase MIC90 of fluconazole (p = 0.05), particularly for C. parapsilosis in the ICU (p = 0.04), with a general and significant decreased trend of the MIC90 values of caspofungin (p = 0.04), and with significant increased MIC50 values for amphotericin B (p = 0.01) over the study period. In conclusion, drug utilization in our hospital turned out to be a putative influencing factor on the ecology of the species, on the increase in time of the biofilm producing strains and on the Candida antifungal susceptibility profile, thus influencing clinical management.

RevDate: 2019-11-07

Booth WT, Davis RR, Deora R, et al (2019)

Structural mechanism for regulation of DNA binding of BpsR, a Bordetella regulator of biofilm formation, by 6-hydroxynicotinic acid.

PloS one, 14(11):e0223387 pii:PONE-D-19-17137.

Bordetella bacteria are respiratory pathogens of humans, birds, and livestock. Bordetella pertussis the causative agent of whopping cough remains a significant health issue. The transcriptional regulator, BpsR, represses a number of Bordetella genes relating to virulence, cell adhesion, cell motility, and nicotinic acid metabolism. DNA binding of BpsR is allosterically regulated by interaction with 6-hydroxynicotinic acid (6HNA), the first product in the nicotinic acid degradation pathway. To understand the mechanism of this regulation, we have determined the crystal structures of BpsR and BpsR in complex with 6HNA. The structures reveal that BpsR binding of 6HNA induces a conformational change in the protein to prevent DNA binding. We have also identified homologs of BpsR in other Gram negative bacteria in which the amino acids involved in recognition of 6HNA are conserved, suggesting a similar mechanism for regulating nicotinic acid degradation.

RevDate: 2019-11-07

Habibipour R, Moradi-Haghgou L, A Farmany (2019)

Green synthesis of AgNPs@PPE and its Pseudomonas aeruginosa biofilm formation activity compared to pomegranate peel extract.

International journal of nanomedicine, 14:6891-6899 pii:209912.

Background: Bacteria are able to form biofilm on the biotic and abiotic surfaces which helps to protect themselves from deleterious conditions, predation, desiccation, and exposure to antibacterial substances. About 80% of bacterial infections are caused by those bacteria living in the biofilm. Pseudomonas aeruginosa, a gram-negative, non-fermentative bacillus, and the ubiquitous bacterium is an important opportunistic pathogen notorious for biofilm formation and is remarkably resistant against most antibiotics multiple front-line antibiotics, which significantly contributes to eradication failure. The aim of this paper was to evaluate the anti-biofilm formation activity of Ag@PPEs gainst P. aeruginosa bacteria.

Methods: An aqueous extract of black pomegranate peel was used for the synthesis of silver nanoparticles (AgNPs@PPE). The characteristics, anti-biofilm formation and cell toxicity of AgNPs@PPE were examined in vitro.

Results: Absorbance at λmax 372 nm which is related to the surface plasmon resonance, confirms the AgNPs@PPE formation. XRD pattern showed the face-centered qubic (fcc) crystalline structure of AgNPs. TEM images showed that spherical AgNPs size is ranged between 32 and 85 nm. The AgNPs@PPE showed inhibition effect against P. aeruginosa biofilm formation at 0.1 to 0.5 mg/ml concentrations. Cell toxicity assay showed that at 400 µg/ml, AgNPs@PPE were safe without a significant toxicity in L929 cell line.

Conclusion: These data indicate that co-treatment of PPE and AgNPs@PPE significantly decreased the biofilm formation rate. Furthermore, no significant toxicity of AgNPs@PPE was shown against L929 cell line at 400 µg/ml concentration.

RevDate: 2019-11-07

Alonso-Calleja C, Gómez-Fernández S, Carballo J, et al (2019)

Prevalence, Molecular Typing, and Determination of the Biofilm-Forming Ability of Listeria monocytogenes Serotypes from Poultry Meat and Poultry Preparations in Spain.

Microorganisms, 7(11): pii:microorganisms7110529.

A study was undertaken of the presence of Listeria monocytogenes in 260 samples of poultry meat obtained from retail outlets in northwestern Spain. L. monocytogenes was detected in 20 samples (7.7%). Twenty strains (one strain per positive sample) were characterized. The strains belonged to 10 serotypes: 1/2a (2 strains), 1/2b (2), 1/2c (2), 3a (1), 3b (2), 3c (2), 4a (2), 4b (4), 4c (1), and 4d (2). Cluster analysis (ribotyping; EcoRI) showed a strong genetic relationship between strains isolated from samples coming from different outlets. Ribotyping permitted some isolates of the same serotype to be differentiated, which points to the possible usefulness of this technique in the epidemiological surveillance of L. monocytogenes. All strains formed biofilm on polystyrene, as shown by confocal laser scanning microscopy. The biovolume (between 621.7 ± 36.0 µm3 and 62,984.0 ± 14,888.2 µm3 in the observational field of 14,161 μm2), percentage of surface coverage (from 2.17 ± 0.84% to 94.43 ± 3.97%), roughness (between 0.399 ± 0.052 and 0.830 ± 0.022), and maximum thickness (between 9.00 ± 0.00 µm and 24.00 ± 14.93 µm) of biofilms varied between strains (p < 0.05). These results expand knowledge of the characteristics of L. monocytogenes isolates from poultry.

RevDate: 2019-11-06

Davis RT, PD Brown (2019)

spoT-mediated stringent response influences environmental and nutritional stress tolerance, biofilm formation and antimicrobial resistance in Klebsiella pneumoniae.

APMIS : acta pathologica, microbiologica, et immunologica Scandinavica [Epub ahead of print].

Klebsiella pneumoniae is an important opportunistic pathogen with significant potential for virulence and multidrug resistance. Treatment failure often occurs because the pathogen may couple virulence and drug resistance with the stringent response. This study assessed the role of the spoT gene in environmental and nutritional stress tolerance, exopolysaccharide capsule production and biofilm formation. spoT mutants were constructed using the Lambda Red recombinase technique, and mutant and wild-type (WT) strains were exposed to limiting concentrations of carbon (glucose), phosphate and aminoacid, and environmental stresses of ethanol, salt and heat. Cell viability, capsule production and cell length were assessed as well as the ability to grow biofilm under antibiotic pressure using gentamicin and ceftazidime. spoT mutants were more susceptible to stresses versus WT; the reverse was true for survival during biofilm susceptibility assay (p < 0.05), especially when carbon and phosphate were present. spoT mutants were elongated and lacked a capsule versus WT and non-starved strains. The inability to produce capsule in mutants before and after starvation was likely a general effect of spoT mutation. These data suggest that the spoT-mediated stringent response is important for K. pneumoniae in conditions of nutrient limitation, environmental stress and antimicrobial pressure.

RevDate: 2019-11-06

Tomizawa T, Ishikawa M, Bello-Irizarry SN, et al (2019)

Biofilm producing Staphylococcus epidermidis (RP62A strain) inhibits osseous integration without osteolysis and histopathology in a murine septic implant model.

Journal of orthopaedic research : official publication of the Orthopaedic Research Society [Epub ahead of print].

Despite its presence in orthopaedic infections, Staphylococcus epidermidis's ability to directly induce inflammation and bone destruction is unknown. Thus, we compared a clinical strain of methicillin-resistant biofilm-producing Staphylococcus epidermidis (RP62A) to a highly virulent and osteolytic strain of methicillin-resistant Staphylococcus aureus (USA300) in an established murine implant-associated osteomyelitis model. Bacterial burden was assessed by colony forming units (CFUs), tissue damage was assessed by histology and micro-computer tomography (µCT), biofilm was assessed by scanning electron microscopy (SEM), host gene expression was assessed by quantitative polymerase chain reaction (qPCR), and osseous integration was assessed via biomechanical push-out test. While CFUs were recovered from RP62A contaminated implants and surrounding tissues after 14 days, the bacterial burden was significantly less than USA300-infected tibiae (p<0.001). Additionally, RP62A failed to produce any of the gross pathologies induced by USA300 (osteolysis, reactive bone-formation, Staphylococcus abscess communities, marrow necrosis and biofilm). However, fibrous tissue was present at the implant-host interface, and rigorous SEM confirmed the rare presence of cocci on RP62A-contaminated implants. Gene expression studies revealed that IL-1β, IL-6, RANKL, and TLR-2 mRNA levels in RP62A infected bone were increased versus Sterile controls. Ex vivo push-out testing showed RP62A infected implants required significantly less force compared to the Sterile group (7.5±3.4 N vs. 17.3±4.1 N; p<0.001), but required 10-fold greater force than USA300 infected implants (0.7±0.3 N; p<0.001). Taken together, these findings demonstrate that S. epidermidis is a commensal pathogen whose mechanisms to inhibit osseous integration are limited to minimal biofilm formation on the implant, and low-grade inflammation. This article is protected by copyright. All rights reserved.

RevDate: 2019-11-06

Majumdar M, Misra TK, DN Roy (2019)

In vitro anti-biofilm activity of 14-deoxy-11,12-didehydroandrographolide from Andrographis paniculata against Pseudomonas aeruginosa.

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] pii:10.1007/s42770-019-00169-0 [Epub ahead of print].

14-Deoxy-11,12-didehydroandrographolide is a biologically active molecule present in the extract of Andrographis paniculata (Kalmegh), a classic ethnic herbal formula, which has been used for over thousand years as therapeutics to treat numerous infectious diseases like upper respiratory tract infection, urinary tract infection, and many more health issues. The present study is designed to ascertain an inhibitor against biofilm formation from the major metabolites of Andrographis paniculata, because the extract of this herb shows inhibition of bacterial quorum sensing (QS) communication and biofilm development against microorganisms. 14-Deoxy-11,12-didehydroandrographolide at 0.1 mM (sub-MIC dose) with azithromycin (6 μg/mL, sub-MIC) or gentamicin (4 μg/mL, sub-MIC) synergistically inhibits 92% biofilm production by a 48-h treatment against Pseudomonas aeruginosa. Further investigation carried out by atomic force microscopy shows promising reduction in roughness and height of biofilm in the presence of 14-deoxy-11,12-didehydroandrographolide compared with the control group. The content of extracellular polymeric substances, level of pyocyanin production, and synthesis of extracellular protease by P. aeruginosa have also been reduced significantly at around 90% in 14-deoxy-11,12-didehydroandrographolide-treated group. In conclusion, 14-deoxy-11,12-didehydroandrographolide could be used as a drug molecule against biofilm development by inhibiting QS pathway in Pseudomonas aeruginosa.

RevDate: 2019-11-06

Brahma U, Sharma P, Murthy S, et al (2019)

Decreased expression of femXAB genes and fnbp mediated biofilm pathways in OS-MRSA clinical isolates.

Scientific reports, 9(1):16028 pii:10.1038/s41598-019-52557-z.

Methicillin-Resistant Staphylococcus aureus (MRSA) is a significant threat to human health. Additionally, biofilm forming bacteria becomes more tolerant to antibiotics and act as bacterial reservoir leading to chronic infection. In this study, we characterised the antibiotic susceptibility, biofilm production and sequence types (ST) of 74 randomly selected clinical isolates of S. aureus causing ocular infections. Antibiotic susceptibility revealed 74% of the isolates as resistant against one or two antibiotics, followed by 16% multidrug-resistant isolates (MDR), and 10% sensitive. The isolates were characterized as MRSA (n = 15), Methicillin-sensitive S. aureus (MSSA, n = 48) and oxacillin susceptible mecA positive S. aureus (OS-MRSA, n = 11) based on oxacillin susceptibility, mecA gene PCR and PBP2a agglutination test. All OS-MRSA would have been misclassified as MSSA on the basis of susceptibility test. Therefore, both phenotypic and genotypic tests should be included to prevent strain misrepresentation. In addition, in-depth studies for understanding the emerging OS-MRSA phenotype is required. The role of fem XAB gene family has been earlier reported in OS-MRSA phenotype. Sequence analysis of the fem XAB genes revealed mutations in fem × (K3R, H11N, N18H and I51V) and fem B (L410F) genes. The fem XAB genes were also found down-regulated in OS-MRSA isolates in comparison to MRSA. In OS-MRSA isolates, biofilm formation is regulated by fibronectin binding proteins A & B. Molecular typing of the isolates revealed genetic diversity. All the isolates produced biofilm, however, MRSA isolates with strong biofilm phenotype represent a worrisome situation and may even result in treatment failure.

RevDate: 2019-11-05

Demir C, Demirci M, Yigin A, et al (2019)

Presence of Biofilm and Adhesin Genes in Staphylococcus aureus Strains Taken from Chronic Wound Infections and their Genotypic and Phenotypic Antimicrobial Sensitivity Patterns.

Photodiagnosis and photodynamic therapy pii:S1572-1000(19)30519-8 [Epub ahead of print].

The purpose of this research was to examine some biofilm (icaA, icaD and bap) and some adhesion (clfA, fnbA, cna) genes, and also assess the genotypic and phenotypic antimicrobial resistance patterns of Staphylococcus aureus strains taken from wound specimens in Mardin, Turkey. A total of 220 wound specimens were investigated. The biofilm forming ability and resistance pattern for eleven antimicrobial agents were investigated by conventional and multiplex PCR methods. S. aureus were taken from 112 (50.9%) of 220 wound specimens. Moreover, biofilm production was found in 79 (70.5%) of the 112 S. aureus isolates. 97 (86.6%) strains of all isolates were positive for icaA and icaD, and 15 (13.4%) for bap. The adhesin genes, cna, fnbA and clfA were detected in 98 (87.5%), 87 (77.7%), and 75 (66.9%) strains, respectively. The numbers of MSSA and MRSA bearing antimicrobial resistance genes were 19 (16.96%) and 32 (28.57%) for blaZ, 9 (8.04%) and 17 (15.18%) for tetK, 6 (5.36%) and 14 (12.5%) for ermC, 2 (1.79%) and 7 (6.25%) for tetM, 0 (0%) and 5 (4.46%) for mecA, 2 (1.79%) and 4 (3.57%) for ermA, 1 (0.89%) and 2 (1.79 %) for both tetK and tetM, respectively. Our findings indicate that multiplex PCR is a suitable way for identifying biofilm and adhesin producing S. aureus. Our data also provided a country-wide oversight of the S. aureus antimicrobial resistance gene profiles for the properly therapy of patients and to control the spreading of the resistance genes.

RevDate: 2019-11-05

Wang T, Flint S, J Palmer (2019)

Magnesium and calcium ions: roles in bacterial cell attachment and biofilm structure maturation.

Biofouling [Epub ahead of print].

The ubiquitous divalent cations magnesium and calcium are important nutrients required by bacteria for growth and cell maintenance. Multi-faceted roles are shown both in bacterial initial attachment and biofilm maturation. The effects of calcium and magnesium can be highlighted in physio-chemical interactions, gene regulation and bio-macromolecular structural modification, which lead to either promotion or inhibition of biofilms. This review outlines recent research addressing phenotypic changes and mechanisms undertaken by calcium and magnesium in affecting bacterial biofilm formation.

RevDate: 2019-11-05

Lauer Cruz K, A de Souza da Motta (2019)

Characterization of biofilm production by Pseudomonas fluorescens isolated from refrigerated raw buffalo milk.

Journal of food science and technology, 56(10):4595-4604.

Pseudomonas fluorescens can often be isolated from refrigerated raw milk. Two strains of P. fluorescens PL5.4 and PL7.1, isolated from raw buffalo milk, were evaluated for their proteolytic capacity, exopolysaccharide production and biofilm production. Proteolytic activity was observed in both strains. The P. fluorescens PL5.4 strain presented fluorescence in the presence of calcofluor, indicating exopolysaccharide production. Both strains were able to produce biofilm at 7 °C for 72 h. For the biofilm production test on stainless steel, adherent cell counts of up to 7.1, 7.3 and 8.8 log CFU/cm2 at 7, 23 and 30 °C were obtained. Through scanning electron microscopy, it was possible to observe the biofilm produced by the P. fluorescens PL5.4 strain. Proper cleaning and disinfection practices in order are important to reduce bacterial contamination and extend the useful life of raw material and its derivatives.

RevDate: 2019-11-05

Yi J, Zhang D, Cheng Y, et al (2019)

The impact of Paenibacillus polymyxa HY96-2 luxS on biofilm formation and control of tomato bacterial wilt.

Applied microbiology and biotechnology pii:10.1007/s00253-019-10162-0 [Epub ahead of print].

The focus of this study was to investigate the effects of luxS, a key regulatory gene of the autoinducer-2 (AI-2) quorum sensing (QS) system, on the biofilm formation and biocontrol efficacy against Ralstonia solanacearum by Paenibacillus polymyxa HY96-2. luxS mutants were constructed and assayed for biofilm formation of the wild-type (WT) strain and luxS mutants of P. polymyxa HY96-2 in vitro and in vivo. The results showed that luxS positively regulated the biofilm formation of HY96-2. Greenhouse experiments of tomato bacterial wilt found that from the early stage to late stage postinoculation, the biocontrol efficacy of the luxS deletion strain was the lowest with 50.70 ± 1.39% in the late stage. However, the luxS overexpression strain had the highest biocontrol efficacy with 75.66 ± 1.94% in the late stage. The complementation of luxS could restore the biocontrol efficacy of the luxS deletion strain with 69.84 ± 1.09% in the late stage, which was higher than that of the WT strain with 65.94 ± 2.73%. Therefore, we deduced that luxS could promote the biofilm formation of P. polymyxa HY96-2 and further promoted its biocontrol efficacy against R. solanacearum.

RevDate: 2019-11-05

Noirot-Gros MF, Forrester S, Malato G, et al (2019)

CRISPR interference to interrogate genes that control biofilm formation in Pseudomonas fluorescens.

Scientific reports, 9(1):15954 pii:10.1038/s41598-019-52400-5.

Bacterial biofilm formation involves signaling and regulatory pathways that control the transition from motile to sessile lifestyle, production of extracellular polymeric matrix, and maturation of the biofilm 3D structure. Biofilms are extensively studied because of their importance in biomedical, ecological and industrial settings. Gene inactivation is a powerful approach for functional studies but it is often labor intensive, limiting systematic gene surveys to the most tractable bacterial hosts. Here, we adapted the CRISPR interference (CRISPRi) system for use in diverse strain isolates of P. fluorescens, SBW25, WH6 and Pf0-1. We found that CRISPRi is applicable to study complex phenotypes such as cell morphology, motility and biofilm formation over extended periods of time. In SBW25, CRISPRi-mediated silencing of genes encoding the GacA/S two-component system and regulatory proteins associated with the cylic di-GMP signaling messenger produced swarming and biofilm phenotypes similar to those obtained after gene inactivation. Combined with detailed confocal microscopy of biofilms, our study also revealed novel phenotypes associated with extracellular matrix biosynthesis as well as the potent inhibition of SBW25 biofilm formation mediated by the PFLU1114 operon. We conclude that CRISPRi is a reliable and scalable approach to investigate gene networks in the diverse P. fluorescens group.

RevDate: 2019-11-05

Adnan M, Ali Shah MR, Jamal M, et al (2019)

Isolation and characterization of bacteriophage to control multidrug-resistant Pseudomonas aeruginosa planktonic cells and biofilm.

Biologicals : journal of the International Association of Biological Standardization pii:S1045-1056(19)30109-5 [Epub ahead of print].

Pseudomonas aeruginosa is Gram-negative bacterium, one of the leading cause of drug-resistant nosocomial infections in developing countries. This bacterium possesses chromosomally encoded efflux pumps, poor permeability of outer-membrane and high tendency for biofilm formation which are tools to confer resistance. Bacteriophages are regarded as feasible treatment option for control of resistant P. aeruginosa. The aim of the current study was isolate and characterized a bacteriophage against P. aeruginosa with MDR and biofilm ability. A bacteriophage MA-1 with moderate host range was isolated from waste water. The phage was considerable heat and pH stable. Electron microscopy revealed that phage MA-1 belongs to Myoviridae family. Its genome was dsDNA (≈50 kb), coding for eighteen different proteins (ranging from 12 to 250 KDa). P. aeruginosa-2949 log growth phase was significantly reduced by phage MA-1 (2.5 × 103 CFU/ml) as compared to control (without phage). Phage MA-1 also showed significant reductions of 2.0, 2.5 and 3.2 folds in 24, 48, and 74 h old biofilms after 6 h treatment with phage respectively as compared to control. It was concluded from this study that phage MA-1 has capability of killing P. aeruginosa planktonic cells and biofilm, but for complete eradication cocktail will more effective to avoid resistance.

RevDate: 2019-11-05

Ribeiro MM, Graziano KU, Olson N, et al (2019)

The polytetrafluoroethylene (PTFE) channel model of cyclic-buildup biofilm and traditional biofilm: The impact of friction, and detergent on cleaning and subsequent high-level disinfection.

Infection control and hospital epidemiology pii:S0899823X19003064 [Epub ahead of print].

OBJECTIVE: To evaluate the efficacy of detergent and friction on removal of traditional biofilm and cyclic-buildup biofilm (CBB) from polytetrafluoroethylene (PTFE) channels and to evaluate the efficacy of glutaraldehyde to kill residual bacteria after cleaning.

METHODS: PTFE channels were exposed to artificial test soil containing 108 CFU/mL of Pseudomonas aeruginosa and Enterococcus faecalis, followed by full cleaning and high-level disinfection (HLD) for five repeated rounds to establish CBB. For traditional biofilm, the HLD step was omitted. Cleaning with enzymatic and alkaline detergents, bristle brush, and Pull Thru channel cleaner were compared to a water flush only. Carbohydrate, protein, viable count, adenosine triphosphate (ATP) levels were analyzed and atomic force microscopy (AFM) was performed.

RESULTS: In the absence of friction, cleaning of traditional biofilm and CBB was not effective compared to the positive control (Dunn-Bonferroni tests; P > .05) regardless of the detergent used. ATP, protein, and carbohydrate analyses were unable to detect traditional biofilm or CBB. The AFM analysis showed that fixation resulted in CBB being smoother and more compact than traditional biofilm.

CONCLUSION: Friction during the cleaning process was a critical parameter regardless of the detergent used for removal of either traditional biofilm or CBB. Glutaraldehyde effectively killed the remaining microorganisms regardless of the cleaning method used.

RevDate: 2019-11-05

Lade H, Park JH, Chung SH, et al (2019)

Biofilm Formation by Staphylococcus aureus Clinical Isolates is Differentially Affected by Glucose and Sodium Chloride Supplemented Culture Media.

Journal of clinical medicine, 8(11): pii:jcm8111853.

Staphylococcus aureus (S. aureus) causes persistent biofilm-related infections. Biofilm formation by S. aureus is affected by the culture conditions and is associated with certain genotypic characteristics. Here, we show that glucose and sodium chloride (NaCl) supplementation of culture media, a common practice in studies of biofilms in vitro, influences both biofilm formation by 40 S. aureus clinical isolates (methicillin-resistant and methicillin-sensitive S. aureus) and causes variations in biofilm quantification. Methicillin-resistant strains formed more robust biofilms than methicillin-sensitive strains in tryptic soy broth (TSB). However, glucose supplementation in TSB greatly promoted and stabilized biofilm formation of all strains, while additional NaCl was less efficient in this respect and resulted in significant variation in biofilm measurements. In addition, we observed that the ST239-SCCmec (Staphylococcal Cassette Chromosome mec) type III lineage formed strong biofilms in TSB supplemented with glucose and NaCl. Links between biofilm formation and accessory gene regulator (agr) status, as assessed by δ-toxin production, and with mannitol fermentation were not found. Our results show that TSB supplemented with 1.0% glucose supports robust biofilm production and reproducible quantification of S. aureus biofilm formation in vitro, whereas additional NaCl results in major variations in measurements of biofilm formation.

RevDate: 2019-11-05

Guimerà X, Moya A, Dorado AD, et al (2019)

A Minimally Invasive Microsensor Specially Designed for Simultaneous Dissolved Oxygen and pH Biofilm Profiling.

Sensors (Basel, Switzerland), 19(21): pii:s19214747.

A novel sensing device for simultaneous dissolved oxygen (DO) and pH monitoring specially designed for biofilm profiling is presented in this work. This device enabled the recording of instantaneous DO and pH dynamic profiles within biofilms, improving the tools available for the study and the characterization of biological systems. The microsensor consisted of two parallel arrays of microelectrodes. Microelectrodes used for DO sensing were bare gold electrodes, while microelectrodes used for pH sensing were platinum-based electrodes modified using electrodeposited iridium oxide. The device was fabricated with a polyimide (Kapton®) film of 127 µm as a substrate for minimizing the damage caused on the biofilm structure during its insertion. The electrodes were covered with a Nafion® layer to increase sensor stability and repeatability and to avoid electrode surface fouling. DO microelectrodes showed a linear response in the range 0-8 mg L-1, a detection limit of 0.05 mg L-1, and a sensitivity of 2.06 nA L mg-1. pH electrodes showed a linear super-Nernstian response (74.2 ± 0.7 mV/pH unit) in a wide pH range (pH 4-9). The multi-analyte sensor array was validated in a flat plate bioreactor where simultaneous and instantaneous pH and DO profiles within a sulfide oxidizing biofilm were recorded. The electrodes spatial resolution, the monitoring sensitivity, and the minimally invasive features exhibited by the proposed microsensor improved biofilm monitoring performance, enabling the quantification of mass transfer resistances and the assessment of biological activity.

RevDate: 2019-11-04

Nisbett LM, Binnenkade L, Bacon BA, et al (2019)

NosP signaling modulates the NO/H-NOX-mediated multicomponent c-di-GMP network and biofilm formation in Shewanella oneidensis.

Biochemistry [Epub ahead of print].

Biofilms form when bacteria aggregate in a self-secreted exopolysaccharide matrix. Biofilms are resistant to antibiotics and thus implicated in human disease. Nitric oxide (NO) is known to mediate biofilm formation in many bacteria via ligation to H-NOX (heme-NO/oxygen binding) proteins. Most NO-responsive bacteria, however, lack H-NOX domain-containing proteins. Our lab has identified another NO-sensing protein (NosP), which is predicted to be involved in two-component signaling networks that regulate biofilm formation in many bacterial species. Here we demonstrate that NosP participates in the previously described H-NOX/NO-responsive multicomponent c-di-GMP signaling network in Shewanella oneidensis. Strains lacking either nosP or its co-cistronic histidine kinase nahK (previously called hnoS) produce immature biofilms, while hnoX and hnoK (histidine kinase responsive to NO/H-NOX) mutants result in wild type biofilm architecture. We demonstrate that NosP regulates the autophosphorylation activity of NahK as well as HnoK. HnoK and NahK have been shown to control the activities of three response regulator proteins (HnoB, HnoC, and HnoD) that together comprise a NO-responsive multicomponent c-di-GMP signaling network. Based on the data presented here, we propose that NosP/NahK adds a level of regulation on top of H-NOX/HnoK to modulate this c-di-GMP signaling network, and ultimately provide temporal regulation of biofilm formation, by governing phosphate flux through both the HnoK and NahK two-component kinases. Further, it appears that NosP and H-NOX each act to counter the activity of the other in a push-pull mechanism; NosP/NahK promotes biofilm formation through inhibition of H-NOX/HnoK signaling, which on its own reduces biofilm formation. Addition of NO to this system results in a reduction of c-di-GMP concentration and biofilm formation, primarily through disinhibition of HnoK activity.

RevDate: 2019-11-04

Kumari P, Arora N, Chatrath A, et al (2019)

Delineating the Biofilm Inhibition Mechanisms of Phenolic and Aldehydic Terpenes against Cryptococcus neoformans.

ACS omega, 4(18):17634-17648.

The recalcitrant biofilm formed by fungus Cryptococcus neoformans is a life-threatening pathogenic condition responsible for further intensifying cryptococcosis. Considering the enhanced biofilm resistance and toxicity of synthetic antifungal drugs, the search for efficient, nontoxic, and cost-effective natural therapeutics has received a major boost. Phenolic (thymol and carvacrol) and aldehydic (citral) terpenes are natural and safe alternatives capable of efficient microbial biofilm inhibition. However, the biofilm inhibition mechanism of these terpenes still remains unclear. In this study, we adopted an integrative biophysical and biochemical approach to elucidate the hierarchy of their action against C. neoformans biofilm cells. The microscopic analysis revealed disruption of the biofilm cell surface with elevation in surface roughness and reduction in cell height. Although all terpenes acted through ergosterol biosynthesis inhibition, the phenolic terpenes also selectively interacted via ergosterol binding. Further, the alterations in the fatty acid profile in response to terpenes attenuated the cell membrane fluidity with enhanced permeability, resulting in pore formation and efflux of the K+/intracellular content. Additionally, mitochondrial depolarization caused higher levels of reactive oxygen species, which led to increased lipid peroxidation and activation of the antioxidant defense system. Indeed, the oxidative stress caused a significant decline in the amount of extracellular polymeric matrix and capsule sugars (mannose, xylose, and glucuronic acid), leading to a reduced capsule size and an overall negative charge on the cell surface. This comprehensive data revealed the mechanistic insights into the mode of action of terpenes on biofilm inhibition, which could be exploited for formulating novel anti-biofilm agents.

RevDate: 2019-11-04

Swetha TK, Pooranachithra M, Subramenium GA, et al (2019)

Umbelliferone Impedes Biofilm Formation and Virulence of Methicillin-Resistant Staphylococcus epidermidis via Impairment of Initial Attachment and Intercellular Adhesion.

Frontiers in cellular and infection microbiology, 9:357.

Staphylococcus epidermidis is an opportunistic human pathogen, which is involved in numerous nosocomial and implant associated infections. Biofilm formation is one of the prime virulence factors of S. epidermidis that supports its colonization on biotic and abiotic surfaces. The global dissemination of three lineages of S. epidermidis superbugs highlights its clinical significance and the imperative need to combat its pathogenicity. Thus, in the current study, the antibiofilm activity of umbelliferone (UMB), a natural product of the coumarin family, was assessed against methicillin-resistant S. epidermidis (MRSE). UMB exhibited significant antibiofilm activity (83%) at 500 μg/ml concentration without growth alteration. Microscopic analysis corroborated the antibiofilm potential of UMB and unveiled its potential to impair intercellular adhesion, which was reflected in auto-aggregation and solid phase adherence assays. Furthermore, real time PCR analysis revealed the reduced expression of adhesion encoding genes (icaD, atlE, aap, bhp, ebh, sdrG, and sdrF). Down regulation of agrA and reduced production of secreted hydrolases upon UMB treatment were speculated to hinder invasive lifestyle of MRSE. Additionally, UMB hindered slime synthesis and biofilm matrix components, which were believed to augment antibiotic susceptibility. In vivo assays using Caenorhabditis elegans divulged the non-toxic nature of UMB and validated the antibiofilm, antivirulence, and antiadherence properties of UMB observed in in vitro assays. Thus, UMB impairs MRSE biofilm by turning down the initial attachment and intercellular adhesion. Altogether, the obtained results suggest the potent antibiofilm activity of UMB and the feasibility of using it in clinical settings for combating S. epidermidis infections.

RevDate: 2019-11-04

Nakagami G, Schultz G, Kitamura A, et al (2019)

Rapid detection of biofilm by wound blotting following sharp debridement of chronic pressure ulcers predicts wound healing: A preliminary study.

International wound journal [Epub ahead of print].

For optimal wound bed preparation, wound debridement is essential to eliminate bacterial biofilms. However, it is challenging for clinicians to determine whether the biofilm is completely removed. A newly developed biofilm detection method based on wound blotting technology may be useful. Thus, we aimed to investigate the effect of biofilm elimination on wound area decrease in pressure ulcers, as confirmed using the wound blotting method. In this retrospective observational study, we enrolled patients with pressure ulcers who underwent sharp debridement with pre- and post-debridement wound blotting. Biofilm was detected on the nitrocellulose membrane using ruthenium red or alcian blue staining. Patients were included if the test was positive for biofilm before wound debridement. Percent decrease in wound area after 1 week was calculated as an outcome measure. We classified the wounds into a biofilm-eliminated group and a biofilm-remaining group based on the post-debridement wound blotting result. Sixteen wound blotting samples from nine pressure ulcers were collected. The percent decrease in wound area was significantly higher in the biofilm-eliminated group (median: 14.4%, interquartile range: 4.6%-20.1%) than in the biofilm-remaining group (median: -14.5%, interquartile range: -25.3%-9.6%; P = .040). The presence of remaining biofilms was an independent predictor for reduced percent decrease in wound area (coefficient = -22.84, P = .040). Biofilm-based wound care guided by wound blotting is a promising measure to help clinicians eliminate bacterial bioburden more effectively for wound area reduction.

RevDate: 2019-11-04

Røder HL, Liu W, Sørensen SJ, et al (2019)

Interspecies interactions reduce selection for a biofilm optimized variant in a four-species biofilm model.

Environmental microbiology reports [Epub ahead of print].

Multispecies biofilms are structured and spatially defined communities, where interspecies interactions impact assembly and functionality. Here we compared the spatial organization and growth of bacterial cells in differently composed biofilm communities over time to determine links between interspecies interactions and selection for biofilm phenotypes of individual species. An established model community consisting of Stenotrophomonas rhizophila, Xanthomonas retroflexus, Microbacterium oxydans and Paenibacillus amylolyticus was used. It was found that interspecies interactions led to varying levels of selection for a new colony phenotype of X. retroflexus, depending on the presence/absence of other species. When M. oxydans was absent, X. retroflexus was not able to establish in the top layers of the biofilm, which led to selection for a hyper-matrix forming phenotype of X. retroflexus that successfully established in the biofilm top layers. No such phenotypic X. retroflexus variants were identified in the presence of M. oxydans. These findings indicate that interspecies interactions may lead to favourable localization of individual species in a multispecies biofilm and thereby reduce selection for competitive phenotypes. This article is protected by copyright. All rights reserved.

RevDate: 2019-11-03

Gabriel C, Grenho L, Cerqueira F, et al (2019)

Inhibitory Effect of 5-Aminoimidazole-4-Carbohydrazonamides Derivatives Against Candida spp. Biofilm on Nanohydroxyapatite Substrate.

Mycopathologia pii:10.1007/s11046-019-00400-4 [Epub ahead of print].

Candida can adhere and form biofilm on biomaterials commonly used in medical devices which is a key attribute that enhances its ability to cause infections in humans. Furthermore, biomaterial-related infections represent a major therapeutic challenge since Candida biofilms are implicated in antifungal therapies failure. The goals of the present work were to investigate the effect of three 5-aminoimidazole-4-carbohydrazonamides, namely (Z)-5-amino-1-methyl-N'-aryl-1H-imidazole-4-carbohydrazonamides [aryl = phenyl (1a), 4-fluorophenyl (1b), 3-fluorophenyl (1c)], on Candida albicans and Candida krusei biofilm on nanohydroxyapatite substrate, a well-known bioactive ceramic material. To address these goals, both quantitative methods (by cultivable cell numbers) and qualitative evaluation (by scanning electron microscopy) were used. Compounds cytocompatibility towards osteoblast-like cells was also evaluated after 24 h of exposure, through resazurin assay. The three tested compounds displayed a strong inhibitory effect on biofilm development of both Candida species as potent in vitro activity against C. albicans sessile cells. Regarding cytocompatibility, a concentration-dependent effect was observed. Together, these findings indicated that the potent activity of imidazole derivatives on Candida spp. biofilms on nanohydroxyapatite substrate, in particular compound 1c, is worth further investigating.

RevDate: 2019-11-03

Zhang H, Wang H, Jie M, et al (2019)

Performance and microbial communities of different biofilm membrane bioreactors with pre-anoxic tanks treating mariculture wastewater.

Bioresource technology, 295:122302 pii:S0960-8524(19)31532-9 [Epub ahead of print].

The performance of pollutant removals, activated sludge characteristics, and microbial communities of two biofilm membrane bioreactors coupled with pre-anoxic tanks (BF-AO-MBRs) (one using fiber bundle bio-carriers (FB-MBR) and the other using suspended bio-carriers (MB-MBR)) were compared at the salinity between zero and 60 g/L. At all salinities, three bioreactors showed good COD average removal efficiencies (>94.1%), and FB-MBR showed the best TN removal efficiency (90.4% at 30 g/L salinity). Moreover, FB-MBR had the faster process start-up time and better salt shock resistance. At high salinities (30-60 g/L), more extracellular polymeric substances were produced by the BF-AO-MBRs to avoid the penetration of salt and protect the bacterial community. Because of the different attachment patterns of biofilms, the microbial community structure in the FB-MBR exposed to 30 g/L salinity had higher nitrite-oxidizing/ammonia-oxidizing bacteria ratio (6.44) with more abundance of denitrifiers, which contribute to higher TN removal efficiency and lower nitrite accumulation.

RevDate: 2019-11-03

D'Andrea MM, Frezza D, Romano E, et al (2019)

The lytic bacteriophage vB_EfaH_EF1TV, a new member of the Herelleviridae family, disrupts biofilm produced by Enterococcus faecalis clinical strains.

Journal of global antimicrobial resistance pii:S2213-7165(19)30273-5 [Epub ahead of print].

OBJECTIVES: The aim of this study is to characterize a new bacteriophage able to infect Enterococcus faecalis, and to evaluate its ability to disrupt biofilm.

METHODS: The vB_EfaH_EF1TV (EF1TV) host-range was determined by spot test and efficiency of plating using a collection of 15E. faecalis clinical strains. The phage genome was sequenced with a next generation sequencing approach. Anti-biofilm activity was tested by crystal violet method and confocal laser scanning microscopy. Phage-resistant mutants were selected and sequenced to investigate receptors exploited by phage for infection.

RESULTS: EF1TV is a newly discoveredE. faecalis phage which belongs to the Herelleviridae family. EF1TV, whose genome is 98% identical to φEF24C, is characterized by a linear dsDNA genome of 143,507 bp with direct terminal repeats of 1,911 bp. The phage is able to infect E. faecalis and shows also the ability to degrade biofilm produced by strains of this species. The results were confirmed by confocal laser scanning microscopy analyzing the biofilm reduction in the same optical field before and after phage infection.

CONCLUSIONS: The EF1TV phage shows promising features such as an obligatory lytic nature, an anti-biofilm activity and the absence of integration-related proteins, antibiotic resistance determinants and virulence factors, and therefore could be a promising tool for therapeutic applications.

RevDate: 2019-11-03

Rathinam NK, Gorky , Bibra M, et al (2019)

Bioelectrochemical approach for enhancing lignocellulose degradation and biofilm formation in Geobacillus strain WSUCF1.

Bioresource technology pii:S0960-8524(19)31501-9 [Epub ahead of print].

Investigations on microbial electrocatalysis as a strategy for enhancing the rates of substrate utilization leading to enhanced yield of biomass and enhanced biofilm formation are reported. A thermophilic Geobacillus sp. strain WSUCF1 (60 °C), a potential lignocellulose degrading microorganism was used as the electrocatalyst. Glucose, cellulose, and corn stover were used as the feedstocks. The results of this investigation showed that applying the oxidation potential of -0.383 mV (vs PRE) increased the glucose utilization and COD removal by 25.5% and 29.7% respectively. The bioelectrocatalysis strategy also increased the biomass yield by 81.2, 42.1, and 49.5% in the case of systems fed with glucose, cellulose, and corn stover, respectively, when compared with the systems without applied oxidation potential. This is the first work reporting the effects of applied oxidation potential on increasing the rates of degradation of lignocellulosic biomass and enhanced biofilm formation.

RevDate: 2019-11-04

Li H, Song HL, Xu H, et al (2019)

Effect of the coexposure of sulfadiazine, ciprofloxacin and zinc on the fate of antibiotic resistance genes, bacterial communities and functions in three-dimensional biofilm-electrode reactors.

Bioresource technology, 296:122290 pii:S0960-8524(19)31520-2 [Epub ahead of print].

Three-dimensional biofilm electrode reactors (3D-BERs) with high treatment efficiency were constructed to treat wastewater containing sulfadiazine (SDZ) and ciprofloxacin (CIP) coexposure with Zinc (Zn). The results showed that coexposure to target antibiotics and Zn increased the absolute and relative abundances of target antibiotic resistance genes (ARGs). Additionally, the target ARG abundances were higher on cathode of 3D-BER compared with ordinary anaerobic reactor while the abundances of total ARGs were decreased in the effluent. Meanwhile, redundancy analysis results revealed that the composition of bacteria carrying ARGs was greatly influenced in the cathode by the accumulation of Zn and antibiotic, which dominated the changes of ARG abundances. Additionally, ARGs with their host bacteria revealed by network analysis were partially deposited on electrode substrates when being removed from wastewater. Thus, 3D-BER exhibits capability of simultaneously eliminating antibiotic and Zn, and greatly reduces the risks of ARGs spread.

RevDate: 2019-11-02

Kipanga PN, Liu M, Panda SK, et al (2019)

Biofilm inhibiting properties of compounds from the leaves of Warburgia ugandensis Sprague subsp ugandensis against Candida and staphylococcal biofilms.

Journal of ethnopharmacology pii:S0378-8741(19)32684-4 [Epub ahead of print].

Warburgia ugandensis Sprague subspecies ugandensis is a plant widely distributed in Eastern, Central and Southern Africa. In humans, it is used to treat respiratory infections, tooth aches, malaria, skin infections, venereal diseases, diarrhea, fevers and aches.

AIM OF THE STUDY: This study aims to identify the bioactive compounds against clinically important biofilm-forming strains of Candida and staphylococci that are responsible for tissue and implanted device-related infections.

METHODS: Using a bioassay-guided fractionation approach, hexane -, ethanol -, acetone - and water extracts from the leaves of W. ugandensis, their subsequent fractions and isolated compounds were tested against both developing and preformed 24 h-biofilms of Candida albicans SC5314, Candida glabrata BG2 Candida glabrata ATCC 2001, Staphylococcus epidermidis 1457 and Staphylococcus aureus USA 300 using microtiter susceptibility tests. Planktonic cells were also tested in parallel for comparison purposes. Confocal scanning laser microscopy was also used to visualize effects of isolated compounds on biofilm formation.

RESULTS: Warburganal, polygodial and alpha-linolenic acid (ALA) were the major bioactive compounds isolated from the acetone extract of W. ugandensis. For both warburganal and polygodial, the biofilm inhibitory concentration that inhibits 50% of C. albicans developing biofilms (BIC50) was 4.5 ± 1 and 10.8 ± 5 μg/mL respectively. Against S. aureus developing biofilms, this value was 37.9 ± 8 μg/mL and 25 μg/mL with warburganal and ALA respectively. Eradication of preformed 24 h biofilms was also observed. Interestingly, synergy between the sesquiterpenoids and azoles against developing C. albicans biofilms resulted in an approximately ten-fold decrease of the effective concentration required to completely inhibit growth of the biofilms by individual compounds. The hydroxyl group in position C-9 in warburganal was identified as essential for activity against staphylococcal biofilms. We also identified additional promising bioactive sesquiterpenoids; drimenol and drimendiol from the structure-activity relationship (SAR) studies.

CONCLUSIONS: ALA and four sesquiterpenoids: polygodial, warburganal, drimenol and drimendiol, have shown biofilm-inhibitory activity that has not been reported before and is worth following up. These compounds are potential drug candidates to manage biofilm-based infections, possibly in combination with azoles.

RevDate: 2019-11-01

Zhang X, Song Z, Hao Ngo H, et al (2019)

Impacts of typical pharmaceuticals and personal care products on the performance and microbial community of a sponge-based moving bed biofilm reactor.

Bioresource technology, 295:122298 pii:S0960-8524(19)31528-7 [Epub ahead of print].

Four lab-scale moving bed biofilm reactors (MBBRs) were built to treat simulated wastewater containing typical pharmaceuticals and personal care products (PPCPs). The efficiency in removing different PPCPs at different concentrations (1, 2 and 5 mg/L) and their effects on the performance of MBBRs were investigated. Results showed that the average removal efficiencies of sulfadiazine, ibuprofen and carbamazepine were 61.1 ± 8.8%, 74.9 ± 8.8% and 28.3 ± 7.4%, respectively. Compared to the reactor without PPCPs, the total nitrogen (TN) removal efficiency of the reactors containing sulfadiazine, ibuprofen and carbamazepine declined by 21%, 30% and 42%, respectively. Based on the microbial community analysis, increasing the PPCPs concentration within a certain range (<2 mg/L) could stimulate microbial growth and increase microbial diversity yet the diversity reduced when the concentration (5 mg/L) exceeded the tolerance of microorganisms. Furthermore the presence and degradation of different PPCPs resulted in a different kind of microbial community structure in the MBBRs.

RevDate: 2019-11-01

Tomiyama K, Shiiya T, Watanabe K, et al (2019)

Effect of toothpaste containing multiple ions-releasing filler on polymicrobial biofilm regrowth and dentin demineralization.

American journal of dentistry, 32(5):245-250.

PURPOSE: To compare the efficacy of toothpaste containing surface pre-reacted glass-ionomer (S-PRG) filler particles to that of conventional sodium fluoride (NaF) toothpaste for the prevention of dentin demineralization and biofilm regrowth.

METHODS: Bovine root dentin specimens and glass coverslips were used as biofilm growth substrates. To establish biofilms, glass and dentin specimens were incubated for 72 hours in 0.2% sucrose McBain medium inoculated with stimulated saliva from a single donor. Specimens then received a single 5-minute treatment with S-PRG toothpaste, fluoride toothpaste, or sterilized deionized water and were incubated in McBain medium for 120 hours to allow biofilm regrowth. Output parameters during regrowth (72-192 hours) were pH of spent medium, colony-forming unit (CFU) counts of biofilms, and dentin mineral profiles, integrated mineral loss (IML: vol% × µm), and lesion depth (Ld). Treatment group differences were tested by one-way ANOVA followed by Tukey's multiple range test (P< 0.05).

RESULTS: At 144 hours, medium pH was significantly higher in the S-PRG-treated dentin group than in the NaF-treated dentin group. In addition, at 192 hours, the CFU count, IML, and Ld were lower in the S-PRG-treated dentin group than in the NaF-treated dentin group. There were significant differences of pH among dentin groups at 72 hours. Treatment with S-PRG toothpaste markedly inhibited dentin demineralization compared to that with NaF toothpaste.

CLINICAL SIGNIFICANCE: Toothpaste containing multiple ions-releasing filler suppressed bacterial viability and inhibited dentin demineralization.

RevDate: 2019-11-01

Ding XS, Zhao B, An Q, et al (2019)

Role of extracellular polymeric substances in biofilm formation by Pseudomonas stutzeri strain XL-2.

Applied microbiology and biotechnology pii:10.1007/s00253-019-10188-4 [Epub ahead of print].

Pseudomonas stutzeri strain XL-2 exhibited significant performance on biofilm formation. Extracellular polymeric substances (EPS) secreted by strain XL-2 were characterized by colorimetry and Fourier transform infrared (FT-IR) spectroscopy. The biofilm growth showed a strong positive correlation (rP=0.96, P<0.01) to extracellular protein content, but no correlation to exopolysaccharide content. Hydrolyzing the biofilm with proteinase K caused a significant decrease in biofilm growth (t=3.7, P<0.05), whereas the changes in biofilm growth were not significant when the biofilm was hydrolyzed by α-amylase and β-amylase, implying that proteins rather than polysaccharides played the dominant role in biofilm formation. More specifically, confocal laser scanning microscopy (CLSM) revealed that the extracellular proteins were tightly bound to the cells, resulting in the cells with EPS presenting more biofilm promotion protein secondary structures, such as three-turn helices, β-sheet, and α-helices, than cells without EPS. Both bio-assays and quantitative analysis demonstrated that strain XL-2 produced signal molecules of N-acylhomoserine lactones (AHLs) during biofilm formation process. The concentrations of C6-HLS and C6-oxo-HLS were both significantly positively correlated with protein contents (P<0.05). Dosing exogenous C6-HLS and C6-oxo-HLS also resulted in the increase in protein content. Therefore, it was speculated that C6-HLS and C6-oxo-HLS released by strain XL-2 could up-regulate the secretion of proteins in EPS, and thus promote the formation of biofilm.

RevDate: 2019-11-01

Asadian M, Azimi L, Alinejad F, et al (2019)

Molecular Characterization of Acinetobacter baumannii Isolated from Ventilator-Associated Pneumonia and Burn Wound Colonization by Random Amplified Polymorphic DNA Polymerase Chain Reaction and the Relationship between Antibiotic Susceptibility and Biofilm Production.

Advanced biomedical research, 8:58 pii:ABR-8-58.

Background: Multidrug-resistant Acinetobacter baumannii can cause complications in antibiotic therapy and increase the rate of morbidity and mortality in hospitalized patients. Patients with ventilator and burns are two specific groups at high risk for A. baumannii infections. This study aimed to determine antibiotic susceptibility patterns associated with biofilm production in A. baumannii and to assess its molecular epidemiology by random amplified polymorphic DNA polymerase chain reaction (RAPD PCR) in A. baumannii isolated from ventilator-associated pneumonia and burn wound colonization.

Materials and Methods: In this study, 79 isolates of A. baumannii (32 ventilator-associated pneumonia [VAP] 47 burns) were collected in two teaching hospitals in Tehran, Iran, in 2018. Conventional biochemical and microbiological methods were used to identify bacteria. Antibiotic susceptibility was detected by disc diffusion methods according to the Clinical and Laboratory Standards Institute 2018. Tube test was examined for the detection of the biofilm formation rate in collected strains. The most prevalent carbapenemase genes were detected by PCR and molecular typing by RAPD PCR.

Results: All of bacteria were extensively drug-resistant (XDR) except for two isolates. The results of tube test indicated that only 36% of XDR strains were in weak rate of biofilm formation group. Two major clonal genetic groups were found in VAP and burn strains. Oxa-23 was the most prevalent carbapenemase in collected A. baumannii.

Conclusion: The presence of XDR strains of A. baumannii is considerable significant problem in hospitals. Further, similar genetic clonal identified in them indicated the nosocomial infection origin. Hence, these results are very important for control of nosocomial infection committee in health-care systems.

RevDate: 2019-11-01

Brown JL, Johnston W, Delaney C, et al (2019)

Biofilm-stimulated epithelium modulates the inflammatory responses in co-cultured immune cells.

Scientific reports, 9(1):15779 pii:10.1038/s41598-019-52115-7.

The gingival epithelium is a physical and immunological barrier to the microbiota of the oral cavity, which interact through soluble mediators with the immune cells that patrol the tissue at the gingival epithelium. We sought to develop a three-dimensional gingivae-biofilm interface model using a commercially available gingival epithelium to study the tissue inflammatory response to oral biofilms associated with "health", "gingivitis" and "periodontitis". These biofilms were developed by sequential addition of microorganisms to mimic the formation of supra- and sub-gingival plaque in vivo. Secondly, to mimic the interactions between gingival epithelium and immune cells in vivo, we integrated peripheral blood mononuclear cells and CD14+ monocytes into our three-dimensional model and were able to assess the inflammatory response in the immune cells cultured with and without gingival epithelium. We describe a differential inflammatory response in immune cells cultured with epithelial tissue, and more so following incubation with epithelium stimulated by "gingivitis-associated" biofilm. These results suggest that gingival epithelium-derived soluble mediators may control the inflammatory status of immune cells in vitro, and therefore targeting of the epithelial response may offer novel therapies. This multi-cellular interface model, both of microbial and host origin, offers a robust in vitro platform to investigate host-pathogens at the epithelial surface.

RevDate: 2019-11-01

Zhang Q, Chen X, Wu H, et al (2019)

Comparison of Clay Ceramsite and Biodegradable Polymers as Carriers in Pack-bed Biofilm Reactor for Nitrate Removal.

International journal of environmental research and public health, 16(21): pii:ijerph16214184.

In recent years, there is a trend of low C/N ratio in municipal domestic wastewater, which results in serious problems for nitrogen removal from wastewater. The addition of an external soluble carbon source has been the usual procedure to achieve denitrification. However, the disadvantage of this treatment process is the need of a closed, rather sophisticated and costly process control as well as the risk of overdosing. Solid-phase denitrification using biodegradable polymers as biofilm carrier and carbon source was considered as an attractive alternative for biological denitrification. The start-up time of the novel process using PCL (polycaprolactone) as biofilm carrier and carbon source was comparable with that of conventional process using ceramsite as biofilm carrier and acetate as carbon source. Further, the solid-phase denitrification process showed higher nitrogen removal efficiency under shorter hydraulic retention time (HRT) and low carbon to nitrogen (C/N) ratio since the biofilm was firmly attached to the clear pores on the surface of PCL carriers and in this process bacteria that could degrade PCL carriers to obtain electron donor for denitrification was found. In addition, solid-phase denitrification process had a stronger resistance of shock loading than that in conventional process. This study revealed, for the first time, that the physical properties of the biodegradable polymer played a vital role in denitrification, and the different microbial compositions of the two processes was the main reason for the different denitrification performances under low C/N ratio.

RevDate: 2019-11-01

Alshanta OA, Shaban S, Nile CJ, et al (2019)

Candida albicans Biofilm Heterogeneity and Tolerance of Clinical Isolates: Implications for Secondary Endodontic Infections.

Antibiotics (Basel, Switzerland), 8(4): pii:antibiotics8040204.

AIM: Endodontic infections are caused by the invasion of various microorganisms into the root canal system. Candida albicans is a biofilm forming yeast and the most prevalent eukaryotic microorganism in endodontic infections. In this study we investigated the ability of C. albicans to tolerate treatment with standard endodontic irrigants NaOCl (sodium hypochlorite), ethylenediaminetetraacetic acid (EDTA) and a combination thereof. We hypothesized that biofilm formed from a panel of clinical isolates differentially tolerate disinfectant regimens, and this may have implications for secondary endodontic infections.

METHODOLOGY: Mature C. albicans biofilms were formed from 30 laboratory and oral clinical isolates and treated with either 3% NaOCl, 17% EDTA or a sequential treatment of 3% NaOCl followed by 17% EDTA for 5 min. Biofilms were then washed, media replenished and cells reincubated for an additional 24, 48 and 72 h at 37 °C. Regrowth was quantified using metabolic reduction, electrical impedance, biofilm biomass and microscopy at 0, 24, 48 and 72 h.

RESULTS: Microscopic analysis and viability readings revealed a significant initial killing effect by NaOCl, followed by a time dependent significant regrowth of C. albicans, but with inter-strain variability. In contrast to NaOCl, there was a continuous reduction in viability after EDTA treatment. Moreover, EDTA significantly inhibited regrowth after NaOCl treatment, though viable cells were still observed.

CONCLUSIONS: Our results indicate that different C. albicans biofilm phenotypes grown in a non-complex surface topography have the potential to differentially tolerate standard endodontic irrigation protocols. This is the first study to report a strain dependent impact on efficacy of endodontic irrigants. Its suggested that within the complex topography of the root canal, a more difficult antimicrobial challenge, that existing endodontic irrigant regimens permit cells to regrow and drive secondary infections.

RevDate: 2019-10-31

Cockeran R, Dix-Peek T, Dickens C, et al (2019)

Biofilm formation and induction of stress response genes is a common response of several serotypes of the pneumococcus to cigarette smoke condensate.

The Journal of infection pii:S0163-4453(19)30326-3 [Epub ahead of print].

OBJECTIVES: Exposure to cigarette smoke impacts on the virulence of Streptococcus pneumoniae (pneumococcus) by mechanisms including induction of biofilm formation. Most studies, however, have focused on individual strains of the pneumococcus. Accordingly, the current study has investigated the commonality of the pneumococcal stress response to cigarette smoke condensate (CSC), using five different strains of the pathogen.

METHODS: Following exposure to CSC at final concentrations of 80 and 160 µg.mL-1 during a 16 hour incubation period, biofilm formation was measured using a crystal violet-based spectrophotometric procedure. Expression of stress genes seemingly linked to biofilm formation viz. hk11 and rr11 [histidine kinase and response regulator of the two-component regulatory system 11 (TCS11) respectively], cat eff (cation efflux system protein), abc (ATP-binding component of an ATP-binding cassette transporter) and 2005-hyp (hypothetical gene) was measured by sequential extraction of RNA, cDNA synthesis and real-time qPCR.

RESULTS: Exposure of all five strains of the pneumococcus to CSC, resulted in significant biofilm formation, as well as induction of all five test stress genes.

CONCLUSIONS: Augmentation of induction of selective stress genes and biofilm formation are common, possibly linked, responses of various serotypes of the pneumococcus to CSC, favouring both persistence of the pathogen and decreased efficacy of antibiotics.

RevDate: 2019-10-31

Torres G, Vargas K, Sánchez-Jiménez M, et al (2019)

Genotypic and phenotypic characterization of biofilm production by Staphylococcus aureus strains isolated from bovine intramammary infections in Colombian dairy farms.

Heliyon, 5(10):e02535 pii:e02535.

The ability of Staphylococcus aureus to form biofilms is an important virulence factor because this has been associated with persistent bovine intramammary infections. Different mechanisms of biofilm formation have been described in S. aureus; however, the process has been found to be mainly driven by the ica and bap genes. The presence of the ica and bap genes, as well as the biofilm formation in vitro were evaluated in 229 S. aureus strains isolated from bovine milk collected from different regions of Department of Antioquia, Colombia. Three different genotypes grouped into three separate clusters were identified from in vitro assays. Genotype 1 (ica positive and bap negative) was the most prevalent (78.17%), followed by genotype 2 (ica and bap positive) (12.66%) and genotype 0 (ica and bap negative) (9.17%). Biofilm formation was observed in 81.26% of the strains from which 100% of genotype 2 isolates showed biofilm formation. The biofilms formed by genotype 2 isolates were also found to have the highest optical density (>2.4). These results showed that most of the S. aureus strains were capable of biofilm formation, suggesting the virulence potential particularly in bap-positive strains.

RevDate: 2019-10-31

Yung YP, McGill SL, Chen H, et al (2019)

Reverse diauxie phenotype in Pseudomonas aeruginosa biofilm revealed by exometabolomics and label-free proteomics.

NPJ biofilms and microbiomes, 5:31 pii:104.

Microorganisms enhance fitness by prioritizing catabolism of available carbon sources using a process known as carbon catabolite repression (CCR). Planktonically grown Pseudomonas aeruginosa is known to prioritize the consumption of organic acids including lactic acid over catabolism of glucose using a CCR strategy termed "reverse diauxie." P. aeruginosa is an opportunistic pathogen with well-documented biofilm phenotypes that are distinct from its planktonic phenotypes. Reverse diauxie has been described in planktonic cultures, but it has not been documented explicitly in P. aeruginosa biofilms. Here a combination of exometabolomics and label-free proteomics was used to analyze planktonic and biofilm phenotypes for reverse diauxie. P. aeruginosa biofilm cultures preferentially consumed lactic acid over glucose, and in addition, the cultures catabolized the substrates completely and did not exhibit the acetate secreting "overflow" metabolism that is typical of many model microorganisms. The biofilm phenotype was enabled by changes in protein abundances, including lactate dehydrogenase, fumarate hydratase, GTP cyclohydrolase, L-ornithine N(5)-monooxygenase, and superoxide dismutase. These results are noteworthy because reverse diauxie-mediated catabolism of organic acids necessitates a terminal electron acceptor like O2, which is typically in low supply in biofilms due to diffusion limitation. Label-free proteomics identified dozens of proteins associated with biofilm formation including 16 that have not been previously reported, highlighting both the advantages of the methodology utilized here and the complexity of the proteomic adaptation for P. aeruginosa biofilms. Documenting the reverse diauxic phenotype in P. aeruginosa biofilms is foundational for understanding cellular nutrient and energy fluxes, which ultimately control growth and virulence.

RevDate: 2019-10-30

Xu Z, Mandic-Mulec I, Zhang H, et al (2019)

Antibiotic Bacillomycin D Affects Iron Acquisition and Biofilm Formation in Bacillus velezensis through a Btr-Mediated FeuABC-Dependent Pathway.

Cell reports, 29(5):1192-1202.e5.

Bacillus spp. produce a wide range of secondary metabolites, including antibiotics, which have been well studied for their antibacterial properties but less so as signaling molecules. Previous results indicated that the lipopeptide bacillomycin D is a signal that promotes biofilm development of Bacillus velezensis SQR9. However, the mechanism behind this signaling is still unknown. Here, we show that bacillomycin D promotes biofilm development by promoting the acquisition of iron. Bacillomycin D promotes the transcription of the iron ABC transporter FeuABC by binding to its transcription factor, Btr. These actions increase intracellular iron concentration and activate the KinB-Spo0A-SinI-SinR-dependent synthesis of biofilm matrix components. We demonstrate that this strategy is beneficial for biofilm development and competition with the Pseudomonas fluorescens PF-5. Our results unravel an antibiotic-dependent signaling mechanism that links iron acquisition to biofilm development and ecological competition.

RevDate: 2019-10-30

Zhong N, Wu Y, Wang Z, et al (2019)

Monitoring Microalgal Biofilm Growth and Phenol Degradation with Fiber-optic Sensors.

Analytical chemistry [Epub ahead of print].

Simple D-type plastic optical fiber (POF) probes (i.e., sensor, reference, and photochemical probes) were created to accurately monitor the progression and phenol degradation of a Chlorella vulgaris biofilm. The sensor and reference probes were used to monitor the biofilm growth (thickness). The sensor probe, which consisted of a D-shaped POF and Canada balsam doped with GeO2 (CBG) coating, was developed to monitor the biofilm growth and change in the liquid-phase composition and its concentration inside the biofilm. The reference probe, which comprised a D-shaped POF, CBG coating, and glass fiber membrane (to separate the liquids from Chlorella vulgaris), was used to measure the response to changes in the liquid phase. A model was developed to demonstrate the accurate measurement of the biofilm thickness. The photochemical POF probe was coupled with a high-permselectivity phenol polymer membrane to monitor the phenol concentration and analyze the degradation time of 50 mg/L phenol with microalgal biofilms. A fixed relationship was obtained between the biofilm sensor output information and biofilm thickness for a biofilm thickness range of 0-290 µm with a periodic supply of 50 mg/L phenol solution. The highest phenol degradation rate occurred at a biofilm thickness of 191-222 µm. The proposed system can be used to investigate microalgal biomass and can provide a promising avenue for research on renewable resources and pollutant degradation.

RevDate: 2019-10-30

Tsiaprazi-Stamou A, Monfort IY, Romani AM, et al (2019)

The synergistic effect of enzymatic detergents on biofilm cleaning from different surfaces.

Biofouling [Epub ahead of print].

Biofilm growth is a significant source of contamination in the food industry. Enzymes are considered green countermeasures against biofilm formation in the food industry owing to their biodegradability and low toxicity. In this study, the synergistic effect of enzymes was studied against biofilm cleaning from hard surfaces. A mixed-microbial sample was sourced from a meat packaging line and biofilms were grown under high shear conditions on stainless steel and polyethylene surfaces. A model cleaning-in-place (CIP) parallel-plate flow chamber was used for firstly, the enzymatic cleaning and secondly, a disinfection step. The cleaning effectiveness was evaluated in response to different formulations containing non-foaming commercial surfactants among with amylase, protease and lipase at neutral pH. The formulation combining all three enzymes was the most effective, showing a synergy essential for the deformation of biofilm structure and consequently better disinfection of both material surfaces.

RevDate: 2019-10-29

Abbas AF, Al-Saadi AGM, MK Alkhudhairy (2019)

Biofilm Formation and Virulence Determinants of Klebsiella oxytoca Clinical Isolates from Patients with Colorectal Cancer.

Journal of gastrointestinal cancer pii:10.1007/s12029-019-00317-7 [Epub ahead of print].

OBJECTIVE: Biofilm formation has made the therapy of bacterial infections more difficult. The objective our study was assessment of pan-drug-resistant (PDR) Klebsiella oxytoca pathogenicity and virulence factors causing AAHC in patients with colorectal cancer (CRC).

MATERIALS AND METHODS: Among a total of 300 healthy and 300 patients with antibiotic-associated hemorrhagic colitis (AAHC) and CRC, 200 K. oxytoca were identified during May 2015-January 2019. The virulence properties and biofilm formation among the isolates were investigated by phenotypic, PCR, and real-time PCR (RT-qPCR) techniques.

RESULTS: The blaCTX-M1 (20%), blaSHV (11%), blaTEM1 (33%), and AmpC encoding CIT (2%) ESBL genes, carbapenemase-encoding genes blaIM (4%) and blaOXA-48 (2%), and colistin-resistant mcr-1 gene (2.5%) were detected. The virulence-encoding genes including fimA (80%), pilQ (100%), matB (100%), mrkA (80%), and npsB (100%) were amplified. Therefore, PDR K. oxytoca containing adhesins and toxin-encoding genes with ability of biofilm formation causing AAHC and CRC were isolated. There was a significant difference between healthy and patients with CRC regarding the presence of K. oxytoca (p = 00.221).

CONCLUSION: Bacterial enteric pathogens possibly play a role in CRC. Biofilm formation by K. oxytoca strains prevents the efficient infection elimination; therefore, rapid identification and control measure are chief requirements. Additionally, more investigations are necessary with this regard.

RevDate: 2019-10-29

Díaz-Pascual F, Hartmann R, Lempp M, et al (2019)

Breakdown of Vibrio cholerae biofilm architecture induced by antibiotics disrupts community barrier function.

Nature microbiology pii:10.1038/s41564-019-0579-2 [Epub ahead of print].

Bacterial cells in nature are frequently exposed to changes in their chemical environment1,2. The response mechanisms of isolated cells to such stimuli have been investigated in great detail. By contrast, little is known about the emergent multicellular responses to environmental changes, such as antibiotic exposure3-7, which may hold the key to understanding the structure and functions of the most common type of bacterial communities: biofilms. Here, by monitoring all individual cells in Vibrio cholerae biofilms during exposure to antibiotics that are commonly administered for cholera infections, we found that translational inhibitors cause strong effects on cell size and shape, as well as biofilm architectural properties. We identified that single-cell-level responses result from the metabolic consequences of inhibition of protein synthesis and that the community-level responses result from an interplay of matrix composition, matrix dissociation and mechanical interactions between cells. We further observed that the antibiotic-induced changes in biofilm architecture have substantial effects on biofilm population dynamics and community assembly by enabling invasion of biofilms by bacteriophages and intruder cells of different species. These mechanistic causes and ecological consequences of biofilm exposure to antibiotics are an important step towards understanding collective bacterial responses to environmental changes, with implications for the effects of antimicrobial therapy on the ecological succession of biofilm communities.

RevDate: 2019-10-29

Flores-Treviño S, Bocanegra-Ibarias P, Camacho-Ortiz A, et al (2019)

Stenotrophomonas maltophilia biofilm: its role in infectious diseases.

Expert review of anti-infective therapy [Epub ahead of print].

Introduction: Infections caused by the opportunistic Stenotrophomonas maltophilia pathogen in immunocompromised patients are complicated to treat due to antibiotic resistance and the ability of the bacteria to produce biofilm. Areas covered: A MEDLINE/PubMed search was performed of available literature to describe the role of biofilm produced by S. maltophilia in the diseases it causes, including biofilm-influencing factors, the biofilm forming process and composition. The antimicrobial resistance due to S. maltophilia biofilm production and current antibiofilm strategies is also included. Expert opinion: Through the production of biofilm, S. maltophilia strains can easily adhere to the surfaces in hospital settings and aid in its transmission. The biofilm can also cause antibiotic tolerance rendering some of the therapeutic options ineffective, causing setbacks in the selection of an appropriate treatment. Conventional susceptibility tests do not yet offer therapeutic guidelines to treat biofilm-associated infections. Current S. maltophilia biofilm control strategies include natural and synthetic compounds, chelating agents, and commonly prescribed antibiotics. As biofilm age and matrix composition affect the level of antibiotic tolerance, their characterization should be included in biofilm susceptibility testing, in addition to molecular and proteomic analyses. As for now, several commonly recommended antibiotics can be used to treat biofilm-related S. maltophilia infections.

RevDate: 2019-10-29

Thakur D, Govindaraju S, Yun K, et al (2019)

The Synergistic Effect of Zinc Ferrite Nanoparticles Uniformly Deposited on Silver Nanowires for the Biofilm Inhibition of Candida albicans.

Nanomaterials (Basel, Switzerland), 9(10): pii:nano9101431.

Near-monodisperse zinc ferrite nanoparticles (ZnFe2O4 NPs) are synthesized by a co-precipitation method and deposited on the surface of silver nanowires (AgNWs), employing a stepwise solution method. The resulting hybrid nanostructures (ZnFe2O4@AgNWs) show a thin and uniform layer of ZnFe2O4 NPs at an optimum weight ratio of 1:6 between the two component nanostructures. The hybrid nanostructures retain the high crystal quality and phase purity of their constituents. It is demonstrated that the ZnFe2O4@AgNWs hybrid nanostructures are effective at inhibiting the biofilm formation of Candida albicans cells. The biofilm inhibition activity of the hybrid nanostructures is estimated to be more than 50% at a low concentration of 100 µg/mL from both crystal violet assay and XTT assay, which are more than 8-fold higher than those of pure AgNWs and ZnFe2O4 NPs. This greatly enhanced biofilm inhibition activity is attributed to the ZnFe2O4 NPs-carrying membrane penetration by AgNWs and the subsequent interaction between Candida cells and ZnFe2O4 NPs. These results indicate that the ZnFe2O4@AgNWs hybrid nanostructures have great potential as a new type of novel antibiofilm agent.


ESP Quick Facts

ESP Origins

In the early 1990's, Robert Robbins was a faculty member at Johns Hopkins, where he directed the informatics core of GDB — the human gene-mapping database of the international human genome project. To share papers with colleagues around the world, he set up a small paper-sharing section on his personal web page. This small project evolved into The Electronic Scholarly Publishing Project.

ESP Support

In 1995, Robbins became the VP/IT of the Fred Hutchinson Cancer Research Center in Seattle, WA. Soon after arriving in Seattle, Robbins secured funding, through the ELSI component of the US Human Genome Project, to create the original ESP.ORG web site, with the formal goal of providing free, world-wide access to the literature of classical genetics.

ESP Rationale

Although the methods of molecular biology can seem almost magical to the uninitiated, the original techniques of classical genetics are readily appreciated by one and all: cross individuals that differ in some inherited trait, collect all of the progeny, score their attributes, and propose mechanisms to explain the patterns of inheritance observed.

ESP Goal

In reading the early works of classical genetics, one is drawn, almost inexorably, into ever more complex models, until molecular explanations begin to seem both necessary and natural. At that point, the tools for understanding genome research are at hand. Assisting readers reach this point was the original goal of The Electronic Scholarly Publishing Project.

ESP Usage

Usage of the site grew rapidly and has remained high. Faculty began to use the site for their assigned readings. Other on-line publishers, ranging from The New York Times to Nature referenced ESP materials in their own publications. Nobel laureates (e.g., Joshua Lederberg) regularly used the site and even wrote to suggest changes and improvements.

ESP Content

When the site began, no journals were making their early content available in digital format. As a result, ESP was obliged to digitize classic literature before it could be made available. For many important papers — such as Mendel's original paper or the first genetic map — ESP had to produce entirely new typeset versions of the works, if they were to be available in a high-quality format.

ESP Help

Early support from the DOE component of the Human Genome Project was critically important for getting the ESP project on a firm foundation. Since that funding ended (nearly 20 years ago), the project has been operated as a purely volunteer effort. Anyone wishing to assist in these efforts should send an email to Robbins.

ESP Plans

With the development of methods for adding typeset side notes to PDF files, the ESP project now plans to add annotated versions of some classical papers to its holdings. We also plan to add new reference and pedagogical material. We have already started providing regularly updated, comprehensive bibliographies to the ESP.ORG site.


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This is a must read book for anyone with an interest in invasion biology. The full title of the book lays out the author's premise — The New Wild: Why Invasive Species Will Be Nature's Salvation. Not only is species movement not bad for ecosystems, it is the way that ecosystems respond to perturbation — it is the way ecosystems heal. Even if you are one of those who is absolutely convinced that invasive species are actually "a blight, pollution, an epidemic, or a cancer on nature", you should read this book to clarify your own thinking. True scientific understanding never comes from just interacting with those with whom you already agree. R. Robbins

Electronic Scholarly Publishing
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Bellingham, WA 98226

E-mail: RJR8222 @

Papers in Classical Genetics

The ESP began as an effort to share a handful of key papers from the early days of classical genetics. Now the collection has grown to include hundreds of papers, in full-text format.

Digital Books

Along with papers on classical genetics, ESP offers a collection of full-text digital books, including many works by Darwin (and even a collection of poetry — Chicago Poems by Carl Sandburg).


ESP now offers a much improved and expanded collection of timelines, designed to give the user choice over subject matter and dates.


Biographical information about many key scientists.

Selected Bibliographies

Bibliographies on several topics of potential interest to the ESP community are now being automatically maintained and generated on the ESP site.

ESP Picks from Around the Web (updated 07 JUL 2018 )