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29 Sep 2022 at 01:43
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Bibliography on: Biofilm


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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 28392838[PMID] NOT 31293528[PMID] NOT 29372251[PMID] NOT pmcbook NOT ispreviousversion

Citations The Papers (from PubMed®)


RevDate: 2022-09-28

Assery N, Alomeir N, Zeng Y, et al (2022)

The Effect of Er:YAG Laser Treatment on Biofilm Formation on Titanium and Zirconia Disc Surfaces.

Journal of periodontology [Epub ahead of print].

BACKGROUND: Lasers represent a promising method for implant decontamination, but evidence on implant surface changes and subsequent biofilm formation is limited. This study aimed to assess the effect of Er:YAG laser treatment on zirconia and titanium discs, and the differences in biofilm formation as a result of surface alterations.

METHODS: A two-stage (in vitro and in vivo) experiment utilizing Er:YAG laser on titanium and zirconia discs was performed. In vitro, surface alterations, roughness, and elemental-material weight differences following laser treatment were assessed using Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). In vivo, four participants wore custom-made intra-oral stents, embedded with laser-treated and untreated titanium and zirconia discs overnight. Biofilm-coated discs were stained using nucleic acid fluorescence dye and visualized using multiphoton confocal laser scanning microscopy. Biofilm 3D structure, biomass, thickness, and live-to-dead bacteria ratio were assessed.

RESULTS: Both titanium (TiZir) and zirconia (YTZP) discs treated with Er:YAG laser resulted in visual surface alterations, but showed no significant change in average surface roughness (titanium P = 0.53, zirconia P = 0.34) or elemental-material-weight (TiZir, P = 0.98), (YTZP, P = 0.96). No significant differences on biofilm biomass, average thickness and live-to-dead bacteria ratio of laser-treated titanium and zirconia discs, were identified compared to untreated groups (titanium P > 0.05, zirconia P >0.05). Generally, zirconia discs presented with a lower live-to-dead bacteria ratio compared to titanium discs, regardless of laser treatment.

CONCLUSION: Er:YAG laser treatment of titanium and zirconia implant surfaces does not significantly affect surface roughness, elemental-material weight, and early biofilm formation in the oral cavity. This article is protected by copyright. All rights reserved.

RevDate: 2022-09-28

Kreth J, Koo H, PI Diaz (2022)

The functional oral microbiome: Biofilm environment, polymicrobial interactions, and community dynamics.

Molecular oral microbiology, 37(5):165-166.

RevDate: 2022-09-28

Tibbits G, Mohamed A, Gelston S, et al (2022)

Activity of a Hypochlorous Acid-Producing Electrochemical-bandage as Assessed with a Porcine Explant Biofilm Model.

Biotechnology and bioengineering [Epub ahead of print].

The activity of a hypochlorous acid-producing electrochemical bandage (e-bandage) in preventing methicillin-resistant Staphylococcus aureus infection (MRSA) infection and removing biofilms formed by MRSA was assessed using a porcine explant biofilm model. e-Bandages inhibited S. aureus infection (P=0.029) after 12 hours (h) of exposure and reduced 3-day biofilm viable cell counts after 6, 12, and 24 h exposures (P=0.029). Needle-type microelectrodes were used to assess HOCl concentrations in explant tissue as a result of e-bandage treatment; toxicity associated with e-bandage treatment was evaluated. HOCl concentrations in infected and uninfected explant tissue varied between 30 - 80 µM, decreasing with increasing distance from the e-bandage. Eukaryotic cell viability was reduced by an average of 71 and 65% in fresh and day 3-old explants, respectively when compared to explants exposed to non-polarized e-bandages. HOCl e-bandages are a promising technology that can be further developed as an antibiotic-free treatment for wound biofilm infections. This article is protected by copyright. All rights reserved.

RevDate: 2022-09-28

Patel H, Buchad H, D Gajjar (2022)

Pseudomonas aeruginosa persister cell formation upon antibiotic exposure in planktonic and biofilm state.

Scientific reports, 12(1):16151.

Persister cell (PC) is dormant, tolerant to antibiotics, and a transient reversible phenotype. These phenotypes are observed in P. aeruginosa and cause bacterial chronic infection as well as recurrence of biofilm-mediated infection. PC formation requires stringent response and toxin-antitoxin (TA) modules. This study shows the P. aeruginosa PC formation in planktonic and biofilm stages on ceftazidime, gentamicin, and ciprofloxacin treatments. The PC formation was studied using persister assay, flow cytometry using Redox Sensor Green, fluorescence as well as Confocal Laser Scanning Microscopy, and gene expression of stringent response and TA genes. In the planktonic stage, ceftazidime showed a high survival fraction, high redox activity, and elongation of cells was observed followed by ciprofloxacin and gentamicin treatment having redox activity and rod-shaped cells. The gene expression of stringent response and TA genes were upregulated on gentamicin followed by ceftazidime treatment and varied among the isolates. In the biofilm stage, gentamicin and ciprofloxacin showed the biphasic killing pattern, redox activity, gene expression level of stringent response and TA varied across the isolates. Ceftazidime treatment showed higher persister cells in planktonic growth while all three antibiotics were able to induce persister cell formation in the biofilm stage.

RevDate: 2022-09-27

Gingichashvili S, Steinberg D, Sionov RV, et al (2022)

An open-source computational tool for measuring bacterial biofilm morphology and growth kinetics upon one-sided exposure to an antimicrobial source.

Scientific reports, 12(1):16125.

Bacillus subtilis biofilms are well known for their complex and highly adaptive morphology. Indeed, their phenotypical diversity and intra-biofilm heterogeneity make this gram-positive bacterium the subject of many scientific papers on the structure of biofilms. The "robustness" of biofilms is a term often used to describe their level of susceptibility to antimicrobial agents and various mechanical and molecular inhibition/eradication methods. In this paper, we use computational analytics to quantify Bacillus subtilis morphological response to proximity to an antimicrobial source, in the form of the antiseptic chlorhexidine. Chlorhexidine droplets, placed in proximity to Bacillus subtilis macrocolonies at different distances result in morphological changes, quantified using Python-based code, which we have made publicly available. Our results quantify peripheral and inner core deformation as well as differences in cellular viability of the two regions. The results reveal that the inner core, which is often characterized by the presence of wrinkled formations in the macrocolony, is more preserved than the periphery. Furthermore, the paper describes a crescent-shaped colony morphology which occurs when the distance from the chlorhexidine source is 0.5 cm, as well as changes observed in the growth substrate of macrocolonies exposed to chlorhexidine.

RevDate: 2022-09-27

Yan H, Liu C, Yu W, et al (2022)

The aggregate distribution of Pseudomonas aeruginosa on biochar facilitate quorum sensing and biofilm formation.

The Science of the total environment pii:S0048-9697(22)06133-2 [Epub ahead of print].

Biochar when applied into soil, together with soil clay minerals, may provide habitats for soil microbes and shift soil microbial community structure. Although several mechanisms have been proposed to explain the effects of biochar on microbial community, the impact of biochar on quorum sensing (QS) and QS-regulated behavior is poorly understood. In this study, we compared the effects of biochar and three common soil minerals (i.e., montmorillonite, kaolinite, and goethite) on QS and biofilm formation. Pseudomonas aeruginosa PAO1 with complete QS systems was selected as a model organism. Our results showed that biochar and goethite effectively promoted microbial QS and biofilm formation, while montmorillonite and kaolinite posed no significant effect. Live/Dead staining, SEM and density-dependent QS activity indicated that biochar was beneficial to cell viability maintenance and cell aggregations, which improved the efficiency of intercellular communications through QS. QS mutant strain experiments confirmed that biochar enhanced PAO1 biofilm formation by promoting QS. Goethite promoted biofilm formation with a different mechanism that cell debris induced by iron ions and positive charge on goethite surface provided raw materials for bacterial biofilm formation. Our findings provide evidence that the presence of biochar can enhance QS and biofilm formation through a feedforward loop of the QS system. This contributes to better understand biochar-mediated microbial cell to cell communications through QS.

RevDate: 2022-09-27

Sun W, Shi S, Chen J, et al (2022)

Blue Light Signaling Regulates Escherichia coli W1688 Biofilm Formation and l-Threonine Production.

Microbiology spectrum [Epub ahead of print].

Escherichia coli biofilm may form naturally on biotic and abiotic surfaces; this represents a promising approach for efficient biochemical production in industrial fermentation. Recently, industrial exploitation of the advantages of optogenetics, such as simple operation, high spatiotemporal control, and programmability, for regulation of biofilm formation has garnered considerable attention. In this study, we used the blue light signaling-induced optogenetic system Magnet in an E. coli biofilm-based immobilized fermentation system to produce l-threonine in sufficient quantity. Blue light signaling significantly affected the phenotype of E. coli W1688. A series of biofilm-related experiments confirmed the inhibitory effect of blue light signaling on E. coli W1688 biofilm. Subsequently, a strain lacking a blue light-sensing protein (YcgF) was constructed via genetic engineering, which substantially reduced the inhibitory effect of blue light signaling on biofilm. A high-efficiency biofilm-forming system, Magnet, was constructed, which enhanced bacterial aggregation and biofilm formation. Furthermore, l-threonine production was increased from 10.12 to 16.57 g/L during immobilized fermentation, and the fermentation period was shortened by 6 h. IMPORTANCE We confirmed the mechanism underlying the inhibitory effects of blue light signaling on E. coli biofilm formation and constructed a strain lacking a blue light-sensing protein; this mitigated the aforementioned effects of blue light signaling and ensured normal fermentation performance. Furthermore, this study elucidated that the blue light signaling-induced optogenetic system Magnet effectively regulates E. coli biofilm formation and contributes to l-threonine production. This study not only enriches the mechanism of blue light signaling to regulate E. coli biofilm formation but also provides a theoretical basis and feasibility reference for the application of optogenetics technology in biofilm-based immobilized fermentation systems.

RevDate: 2022-09-26

Ensafi F, Fazlyab M, Chiniforush N, et al (2022)

Comparative effects of SWEEPS technique and antimicrobial photodynamic therapy by using curcumin and nano-curcumin on Enterococcus faecalis biofilm in root canal treatment.

Photodiagnosis and photodynamic therapy pii:S1572-1000(22)00416-1 [Epub ahead of print].

OBJECTIVES: This study aimed to compare the antimicrobial effects of the shock wave enhanced emission photoacoustic streaming (SWEEPS) technique and Antimicrobial photodynamic therapy (aPDT) with curcumin and nano-curcumin photosensitizers on Enterococcus faecalis (E. faecalis) biofilm in root canals of extracted teeth.

MATERIALS AND METHODS: This experimental study was conducted on extracted single-rooted human teeth. The teeth were decoronated at the cementoenamel junction, the root canals were instrumented with BioRace system, and their apices were sealed. Smear layer was removed, and the teeth were autoclave-sterilized. E. faecalis strains were cultured in sterile brain heart infusion (BHI) and a bacterial suspension with 0.5 McFarland standard concentration was prepared. The root canals were inoculated with the bacterial suspension and incubated at 37°C for 4 weeks. The teeth were then disinfected in 10 groups (curcumin, nano-curcumin, curcumin + LED, nano-curcumin + LED, curcumin + SWEEPS, nano-curcumin + SWEEPS, curcumin + LED + SWEEPS, nano-curcumin + LED + SWEEPS, and SWEEPS). The number of colony forming units (CFUs) was counted and analyzed by one-way ANOVA and Tukey's test.

RESULTS: A significant difference was noted in colony count among the groups (P<0.001). LED alone had the lowest and nano-curcumin + LED + SWEEPS had the highest root canal cleaning efficacy. SWEEPS along with curcumin, nano-curcumin, and LED significantly decreased the bacterial count compared with other groups (P<0.05).

CONCLUSION: Application of the SWEEPS technique with curcumin and nano-curcumin as photosensitizer activated by LED in aPDT improved the reduction of E. faecalis in root canals.

RevDate: 2022-09-26

Feng W, Yang J, Ma Y, et al (2022)

Correlation between SAP2 and CAP1 in clinical strains of Candida albicans at planktonic and biofilm states.

Canadian journal of microbiology [Epub ahead of print].

This study aimed to explore the influences of SAP2 and CAP1 on itraconazole (ITR) resistance of C. albicans at different states. A total of 10 ITR resistant strains and 10 sensitive strains were used for SAP2 sequencing and CAP1 sequencing. SAP2 sequencing showed no missense mutation, and three synonymous mutations. CAP1 gene sequencing identified two missense mutations (M140I and K191Q), and 14 synonymous mutations. The biofilm formation capacity of resistant C. albicans strains, including the CAP1∆/∆ strain, was stronger. Afterwards, real-time quantitative PCR was used to analyze the expression of SAP2 and CAP1. Compared with the sensitive strains, SAP2 and CAP1 expression were both significantly upregulated in resistant strains at planktonic and biofilm states (P < 0.05). Compared with the strains at planktonic state, SAP2 was significantly upregulated, while CAP1 was significantly downregulated at biofilm states (P < 0.05). Additionally, SAP2 expression in the CAP1 knocked down strain of C. albicans was significantly upregulated, as well as SAP2 expression was evidently downregulated in the CAP1∆/∆ strain at biofilm states compared to that at planktonic states. Loss of CAP1 can increase SAP2 level and may influence the biofilm formation of C. albicans, thus increasing ITR resistance of C. albicans.

RevDate: 2022-09-28

Mosayebi N, Toodehzaeim MH, Zandi H, et al (2022)

Evaluation of the effects of fluoride mouth rinse and varnish on the early biofilm formation of Streptococcus mutans in two types of orthodontic adhesive resins: An in vitro study.

Dental research journal, 19:54.

Background: The aim of the present study is to compare the antibacterial effect of fluoride mouth rinse and fluoride varnish on the primary biofilm formation of Streptococcus mutans (S. mutans) in two types of orthodontic adhesives.

Materials and Methods: This is an in vitro study in which forty composite discs of Transbond XT and Lightbond were divided randomly into 4 groups: Group 1: Control group (not treated with fluoride), Group 2: Rinsed by 0.2% fluoride mouth rinse, Group 3: Rinsed by 0.05% fluoride mouth rinse, and Group 4: Treated by varnish fluoride. Then each group was placed in S. mutans suspension. Bacterial suspension from each treatment was subcultured onto the surface of Mueller-Hinton agar plates, and bacterial growth was assessed. The results were analyzed by analysis of variance test and Scheffé test was run to compute the binary groups (P < 0.05).

Results: There was a statistically significant reduction in the viability of S. mutans in treated groups by fluoride but no significant difference between two types of composites.

Conclusion: The results of this study demonstrated that S. mutans colonies were sensitive to fluoride and their most effective form was varnish. There was no significant difference in early biofilm formation of S. mutans in two types of orthodontic adhesive resins Transbond XT and Lightbond.

RevDate: 2022-09-28
CmpDate: 2022-09-28

Yang SQ, Zhou HJ, Teng LP, et al (2022)

Streptomyces: Derived Active Extract Inhibits Candida albicans Biofilm Formation.

Current microbiology, 79(11):332.

Candida albicans is an opportunistic pathogen that causes biofilm-associated infections. C. albicans biofilms are known to display reduced susceptibility to antimicrobials and high rates of acquired antibiotic resistance, and biofilm forming in C. albicans further hampers treatment options and highlights the need for new antibiofilm strategies. Identifying active components from desert actinomycetes strains to inhibit the formation of C. albicans biofilms represents an effective treatment strategy. In this study, actinomycetes that can inhibit C. albicans biofilm formation were isolated from the Taklimakan Desert, and the underlying mechanisms were explored. After screening the anti-C.albicans biofilm activities of culture supernatants from 170 Actinomycete strains, six strains showed significant inhibition of C. albicans biofilm formation. Microscopic examination showed a reduction in biofilm formation of C. albicans treated with supernatants from actinomycetes. Scanning electron microscopy showed that the morphological changes in biofilm cells were caused by cell membrane rupture and cell material leakage. Then, C.albicans biofilms were destroyed by changing the content of extracellular polysaccharides or degrading extracellular DNA. Finally, a preliminary study on active substances extracted from a new species (TRM43335) showed that the substances that inhibited the formation of biofilms might be peptides. This study provides preliminary evidence that desert actinomyces strains have inhibitory effects on the biofilm development of C. albicans.

RevDate: 2022-09-26

Pesset CM, Fonseca COD, Antunes M, et al (2022)

Characterizing biofilm formation of Staphylococcus pseudintermedius in different suture materials.

Microbial pathogenesis pii:S0882-4010(22)00409-0 [Epub ahead of print].

Staphylococcus pseudintermedius is the primary cause of pyoderma and surgical site infection (SSI) in dogs, and biofilm formation is the main reason for persistent SSI. The presence of biofilm in medical devices can directly impact treatment. Methicillin-resistant S. pseudintermedius (MRSP) emerged rapidly in companion animals, limiting treatment options. MRSP is a public health problem since zoonotic transmission can occur. The study seeks to evaluate biofilm formation capacity via Staphylococcus pseudintermedius collected from dogs affected by topical infections, in suture materials commonly used in companion animal surgery. We tested segments of four types of sutures. Biofilm production was measured by staining with safranin and colorimetric absorbance measurement. We calculated colony-forming units (CFUs) for each type of sutures and visualized biofilm via Scanning Electron Microscopy (SEM) images. The genes associated with biofilm formation (icaA and icaD) were identified using PCR. The colorimetric tests showed that the biofilm is most abundantly formed on the cotton sutures and polyglactin 910. The ability to form biofilm on polypropylene and nylon sutures has also been demonstrated, although at varying intensities. PCR revealed the presence of the two genes (icaA and icaD) in all the isolates. We used a positive control using a reference strain and negative control without bacteria for comparisons. Suture material allowing biofilm formation makes it difficult to prevent and treat surgical site infections. Therefore, it is important to know which suture thread is more susceptible to biofilm formation by bacteria to prevent possible secondary infections at surgical sites.

RevDate: 2022-09-28
CmpDate: 2022-09-28

da Silva NDG, de Paiva PRB, Magalhães TVM, et al (2022)

Effect of experimental and commercial artificial saliva formulations on the activity and viability of microcosm biofilm and on enamel demineralization for irradiated patients with head and neck cancer (HNC).

Biofouling, 38(7):674-686.

The effect of different artificial saliva formulations on biofilm activity and viability, and on enamel demineralization for head and neck cancer (HNC) patients was evaluated. Irradiated enamel samples were treated (1 min) with BioXtra® or with experimental formulations containing carboxymethylcellulose plus inorganic constituents alone (AS) or containing 0.1 mg mL-1 CaneCPI-5 (AS + Cane), 1.0 mg mL-1 hemoglobin (AS + Hb) or combination of both (AS + Cane + Hb). Phosphate-buffered-saline and chlorhexidine (0.12%) were negative and positive control, respectively. Biofilm was produced from the saliva of five male HNC patients, under 0.2% sucrose exposure for 5 days, and daily treated with the formulations (1 min). No significant effects were observed for the different experimental treatments. BioXtra® significantly reduced lactobacilli, demonstrating antibacterial potential for this group. Chlorhexidine was an effective treatment to significantly reduce all parameters, being an important antimicrobial and anticaries agent. Future in vitro studies must be performed using a new approach for the design of the experimental formulations.

RevDate: 2022-09-26

Prentice JA, Bridges AA, BL Bassler (2022)

Synergy between c-di-GMP and Quorum-Sensing Signaling in Vibrio cholerae Biofilm Morphogenesis.

Journal of bacteriology [Epub ahead of print].

Transitions between individual and communal lifestyles allow bacteria to adapt to changing environments. Bacteria must integrate information encoded in multiple sensory cues to appropriately undertake these transitions. Here, we investigate how two prevalent sensory inputs converge on biofilm morphogenesis: quorum sensing, which endows bacteria with the ability to communicate and coordinate group behaviors, and second messenger c-di-GMP signaling, which allows bacteria to detect and respond to environmental stimuli. We use Vibrio cholerae as our model system, the autoinducer AI-2 to modulate quorum sensing, and the polyamine norspermidine to modulate NspS-MbaA-mediated c-di-GMP production. Individually, AI-2 and norspermidine drive opposing biofilm phenotypes, with AI-2 repressing and norspermidine inducing biofilm formation. Surprisingly, however, when AI-2 and norspermidine are simultaneously detected, they act synergistically to increase biofilm biomass and biofilm cell density. We show that this effect is caused by quorum-sensing-mediated activation of nspS-mbaA expression, which increases the levels of NspS and MbaA, and in turn, c-di-GMP biosynthesis, in response to norspermidine. Increased MbaA-synthesized c-di-GMP activates the VpsR transcription factor, driving elevated expression of genes encoding key biofilm matrix components. Thus, in the context of biofilm morphogenesis in V. cholerae, quorum-sensing regulation of c-di-GMP-metabolizing receptor levels connects changes in cell population density to detection of environmental stimuli. IMPORTANCE The development of multicellular communities, known as biofilms, facilitates beneficial functions of gut microbiome bacteria and makes bacterial pathogens recalcitrant to treatment. Understanding how bacteria regulate the biofilm life cycle is fundamental to biofilm control in industrial processes and in medicine. Here, we demonstrate how two major sensory inputs-quorum-sensing communication and second messenger c-di-GMP signaling-jointly regulate biofilm morphogenesis in the global pathogen Vibrio cholerae. We characterize the mechanism underlying a surprising synergy between quorum-sensing and c-di-GMP signaling in controlling biofilm development. Thus, the work connects changes in cell population density to detection of environmental stimuli in a pathogen of clinical significance.

RevDate: 2022-09-24

Zhao H, Shi T, Li F, et al (2022)

[Advance of in vitro oral static biofilm model].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology, 38(9):3267-3278.

Oral static biofilm model is an important tool for in vitro simulation of oral microecological environment, which has become an important method for studying the pathogenesis of various oral diseases and testing the efficacy of various drugs, oral care products and foods due to its low cost, high throughput, good reliability and easy operation. The establishment of oral static biofilm models allows the selection of different devices, inoculum sources, media, substrates and culture conditions according to the purpose of the study, and the evaluation of biofilm growth by various methods such as measuring biomass, metabolic activity, community structure and performing visualization analysis. This paper summarizes the methodological elements reported in recent years for the establishment and evaluation of oral static biofilm models, and analyzes and discusses the applicability of various methods in the hope of contributing to the research and production practice in related fields.

RevDate: 2022-09-24

Wen X, Huang J, Zeng G, et al (2022)

Microbial activity along the depth of biofilm in simultaneous partial nitrification, anammox and denitrification (SNAD) system.

Environmental technology [Epub ahead of print].

Simultaneous partial nitrification, anammox and denitrification (SNAD) is a sustainable and cost-effective technology for nitrogen removal from low strength wastewater. However, knowledge of biofilm microenvironment of SNAD system is currently unsatisfactory. The purpose of this study was to evaluate organic carbon effects on microenvironment and microbial growth in SNAD biofilm system. Microelectrodes were used to investigate microbial activity in-depth within biofilms. ORP distribution of SNAD system was positively related to anammox activity(R2=0.9), and had some influence on microbial community structure. The synergistic effect of anammox bacteria and denitrifiers could be achieved when the abundance ratio of anammox bacteria to denitrifying bacteria is greater than 1.2.

RevDate: 2022-09-23

Haque MM, Haque MA, Mosharaf MK, et al (2022)

Biofilm-mediated decolorization, degradation and detoxification of synthetic effluent by novel biofilm-producing bacteria isolated from textile dyeing effluent.

Environmental pollution (Barking, Essex : 1987) pii:S0269-7491(22)01451-8 [Epub ahead of print].

Biofilm mediated bioremediation of xenobiotic pollutants is an environmental friendly biological technique. In this study, 36 out of 55 bacterial isolates developed biofilms in glass test tubes containing salt-optimized broth plus 2% glycerol (SOBG). Scanning electron microscopy, Fourier transform infrared (FTIR) spectroscopy, and Congo red- and Calcofluor binding results showed biofilm matrices contain proteins, curli, nanocellulose-rich polysaccharides, nucleic acids, lipids, and peptidoglycans. Several functional groups including -OH, N-H, C-H, CO, COO-, -NH2, PO, C-O, and C-C were also predicted. By sequencing, ten novel biofilm-producing bacteria (BPB) were identified, including Exiguobacterium indicum ES31G, Kurthia gibsonii ES43G, Kluyvera cryocrescens ES45G, Cedecea lapagei ES48G, Enterobacter wuhouensis ES49G, Aeromonas caviae ES50G, Lysinibacillus sphaericus ES51G, Acinetobacter haemolyticus ES52G, Enterobacter soli ES53G, and Comamonas aquatic ES54G. The Direct Red (DR) 28 (a carcinogenic and mutagenic dye used in dyeing and biomedical processes) decolorization process was optimized in selected bacterial isolates. Under optimum conditions (SOBG medium, 75 mg L-1 dye, pH 7, 28 °C, microaerophilic condition and within 72 h of incubation), five of the bacteria tested could decolorize 97.8% ± 0.56-99.7% ± 0.45 of DR 28 dye. Azoreductase and laccase enzymes responsible for biodegradation were produced under the optimum condition. UV-Vis spectral analysis revealed that the azo (-NN-) bond peak at 476 nm had almost disappeared in all of the decolorized samples. FTIR data revealed that the foremost characteristic peaks had either partly or entirely vanished or were malformed or stretched. The chemical oxygen demand decreased by 83.3-91.3% in the decolorized samples, while plant probiotic bacterial growth was indistinguishable in the biodegraded metabolites and the original dye. Furthermore, seed germination (%) was higher in the biodegraded metabolites than the parent dye. Thus, examined BPB could provide potential solutions for the bioremediation of industrial dyes in wastewater.

RevDate: 2022-09-23

Lima LS, Ramalho SR, Sandim GC, et al (2022)

Prevention of hospital pathogen biofilm formation by antimicrobial peptide KWI18.

Microbial pathogenesis pii:S0882-4010(22)00404-1 [Epub ahead of print].

This study investigated the antimicrobial and antibiofilm activity of KWI18, a new synthetic peptide. KWI18 was tested against planktonic cells and Pseudomonas aeruginosa and Candida parapsilosis biofilms. Time-kill and synergism assays were performed. Sorbitol, ergosterol, lipid peroxidation, and protein oxidation assays were used to gain insight into the mechanism of action of the peptide. Toxicity was evaluated against erythrocytes and Galleria mellonella. KWI18 showed antimicrobial activity, with minimum inhibitory concentration (MIC) values ranging from 0.5 to 10 μM. KWI18 at 10× MIC reduced P. aeruginosa and C. parapsilosis biofilm formation and cell viability. Time-kill assays revealed that KWI18 inhibited the growth of P. aeruginosa in 4 h and that of C. parapsilosis in 6 h. The mechanism of action was related to ergosterol as well as induction of oxidative damage in cells and biofilms. Furthermore, KWI18 demonstrated low toxicity to erythrocytes and G. mellonella. KWI18 proved to be an effective antibiofilm agent, opening opportunities for the development of new antimicrobials.

RevDate: 2022-09-23

Robberecht L, Delattre J, M Meire (2022)

Isthmus morphology influences debridement efficacy of activated irrigation: a laboratory study involving biofilm mimicking hydrogel removal and high-speed imaging.

International endodontic journal [Epub ahead of print].

AIM: Little is known about the influence of isthmus morphology on the debridement efficacy of activated irrigation. The aim of this study was to investigate the influence of isthmus morphology on the debridement efficacy of laser-activated irrigation (LAI), EDDY and needle irrigation (NI), and to explain the methods of isthmus cleaning by LAI and EDDY.

METHODOLOGY: Four root canal models (apical diameter: 0.30 mm, taper: 0.06, curvature: 23°, length: 20 mm) were produced by CAD-CAM with different isthmus morphologies: long-wide (4 mm; 0.4 mm), long-narrow (4 mm; 0.15 mm), short-wide (2 mm; 0.4 mm) and short-narrow (2 mm; 0.15 mm). The isthmuses were filled with a hydrogel containing dentine debris. The canals were filled with irrigant and models were assigned to the following irrigation protocols (n=240): needle irrigation (NI) with a 30G needle, Eddy, and LAI (2940 nm Er:YAG-laser, 15 Hz, 40 mJ, SWEEPS, tip at the canal entrance). Standardized images of the isthmuses were taken before and after irrigation, and the amount of removed hydrogel was determined using image analysis software and compared across groups using Kruskal-Wallis test followed by Dunn's multiple comparison. Visualization of the isthmus during activation was achieved using a high-speed camera. The pattern and speed of the flow in the isthmus as well as transient and stable cavitation were analysed using imaging software.

RESULTS: LAI, EDDY and NI removed more hydrogel in short-wide isthmuses than in narrow isthmuses (P<0.001). LAI and EDDY removed more hydrogel than NI in every isthmus configuration (P<0.001). EDDY showed eddies and stable cavitation, and LAI showed transient cavitation at each pulse, and pulsed horizontal flow with the highest particle speed in closed short isthmuses.

CONCLUSIONS: Isthmus morphology influences debridement in all irrigation groups. Short-wide isthmuses were the easiest to clean while narrow isthmuses were the most challenging to clean. Width seems to be a more critical anatomical parameter than length. LAI and EDDY resulted in the greatest biofilm removal and performed better than NI. EDDY produced eddies and stable cavitation in the isthmus, and LAI showed transient cavitation and pulsed horizontal flow.

RevDate: 2022-09-23

Wanicharat W, Wanachantararak P, Poomanee W, et al (2022)

Potential of Bouea macrophylla kernel extract as an intracanal medicament against mixed-species bacterial-fungal biofilm. An in vitro and ex vivo study.

Archives of oral biology, 143:105539 pii:S0003-9969(22)00196-0 [Epub ahead of print].

OBJECTIVE: To investigate the antimicrobial activity of B. macrophylla kernel extract against mixed-species biofilms of E. faecalis, S. gordonii and C. albicans in vitro. To evaluate the efficacy of the extract as an intracanal medicament compared with Ca(OH)2 and chlorhexidine in ex vivo tooth model.

METHODS: The antibiofilm effect of B. macrophylla kernel extract was determined by AlamarBlue™ assay and the effect on biofilms was visualized by LIVE/DEAD® BacLight™ viability test. Mixed-species biofilms were incubated into the tooth model (N = 42) for 21 days. The teeth were randomly divided into 4 medicament groups for 7 days: (i) normal saline, (ii) calcium hydroxide (Ca(OH)2), (iii) chlorhexidine gel, (iv) B. macrophylla kernel extract. Dentine samples were collected, qPCR with PMA was used to quantify the viability and species composition of each sample. SEM was used to visualize the effect of medicament on biofilm structure.

RESULTS: The MBIC was 6.25 mg/mL and the MBEC was 50 mg/mL. The integrity of microbial cells was progressively compromised as concentration increased, resulting in greater cell death. Ex vivo tooth model revealed that biofilm treated with 50 mg/mL of the B. macrophylla extract demonstrated a significantly higher proportions of dead cells than in Ca(OH)2, chlorhexidine and normal saline groups (p < 0.01). Disruption of biofilm structure and enlargement of dentinal tubules was observed in B. macrophylla group on SEM.

CONCLUSION: The extract of B. macrophylla kernel exhibited significant antibiofilm effect against the mixed-species biofilms of E. faecalis, S. gordonii and C. albicans.

RevDate: 2022-09-23

Porter M, Davidson FA, MacPhee CE, et al (2022)

Systematic microscopical analysis reveals obligate synergy between extracellular matrix components during Bacillus subtilis colony biofilm development.

Biofilm, 4:100082 pii:S2590-2075(22)00016-8.

Single-species bacterial colony biofilms often present recurring morphologies that are thought to be of benefit to the population of cells within and are known to be dependent on the self-produced extracellular matrix. However, much remains unknown in terms of the developmental process at the single cell level. Here, we design and implement systematic time-lapse imaging and quantitative analyses of the growth of Bacillus subtilis colony biofilms. We follow the development from the initial deposition of founding cells through to the formation of large-scale complex structures. Using the model biofilm strain NCIB 3610, we examine the movement dynamics of the growing biomass and compare them with those displayed by a suite of otherwise isogenic matrix-mutant strains. Correspondingly, we assess the impact of an incomplete matrix on biofilm morphologies and sessile growth rate. Our results indicate that radial expansion of colony biofilms results from the division of bacteria at the biofilm periphery rather than being driven by swelling due to fluid intake. Moreover, we show that lack of exopolysaccharide production has a negative impact on cell division rate, and the extracellular matrix components act synergistically to give the biomass the structural strength to produce aerial protrusions and agar substrate-deforming ability.

RevDate: 2022-09-23

de Almeida OGG, Pereira MG, Oxaran V, et al (2022)

In silico metatranscriptomic approach for tracking biofilm-related effectors in dairies and its importance for improving food safety.

Frontiers in microbiology, 13:928480.

Sessile microorganisms are usually recalcitrant to antimicrobial treatments, and it is possible that finding biofilm-related effectors in metatranscriptomics datasets helps to understand mechanisms for bacterial persistence in diverse environments, by revealing protein-encoding genes that are expressed in situ. For this research, selected dairy-associated metatranscriptomics bioprojects were downloaded from the public databases JGI GOLD and NCBI (eight milk and 45 cheese samples), to screen for sequences encoding biofilm-related effectors. Based on the literature, the selected genetic determinants were related to adhesins, BAP, flagellum-related, intraspecific QS (AHL, HK, and RR), interspecific QS (LuxS), and QQ (AHL-acylases, AHL-lactonases). To search for the mRNA sequences encoding for those effector proteins, a custom database was built from UniprotKB, yielding 1,154,446 de-replicated sequences that were indexed in DIAMOND for alignment. The results revealed that in all the dairy-associated metatranscriptomic datasets obtained, there were reads assigned to genes involved with flagella, adhesion, and QS/QQ, but BAP-reads were found only for milk. Significant Pearson correlations (p < 0.05) were observed for transcripts encoding for flagella, RR, histidine kinases, adhesins, and LuxS, although no other significant correlations were found. In conclusion, the rationale used in this study was useful to demonstrate the presence of biofilm-associated effectors in metatranscriptomics datasets, pointing out to possible regulatory mechanisms in action in dairy-related biofilms, which could be targeted in the future to improve food safety.

RevDate: 2022-09-23

Gu K, Ouyang P, Hong Y, et al (2022)

Geraniol inhibits biofilm formation of methicillin-resistant Staphylococcus aureus and increase the therapeutic effect of vancomycin in vivo.

Frontiers in microbiology, 13:960728.

Methicillin-resistant Staphylococcus aureus (MRSA) is among the common drug resistant bacteria, which has gained worldwide attention due to its high drug resistance and infection rates. Biofilms produced by S. aureus are known to increase antibiotic resistance, making the treatment of S. aureus infections even more challenging. Hence, inhibition of biofilm formation has become an alternative strategy for controlling persistent infections. In this study, we evaluated the efficacy of geraniol as a treatment for MRSA biofilm infection. The results of crystal violet staining indicated that 256 μg/mL concentration of geraniol inhibited USA300 biofilm formation by 86.13% and removed mature biofilms by 49.87%. Geraniol exerted its anti-biofilm effect by influencing the major components of the MRSA biofilm structure. We found that geraniol inhibited the synthesis of major virulence factors, including staphyloxanthin and autolysins. The colony count revealed that geraniol inhibited staphyloxanthin and sensitized USA300 cells to hydrogen peroxide. Interestingly, geraniol not only reduced the release of extracellular nucleic acids (eDNA) but also inhibited cell autolysis. Real-time polymerase chain reaction data revealed the downregulation of genes involved in biofilm formation, which verified the results of the phenotypic analysis. Geraniol increased the effect of vancomycin in eliminating USA300 biofilms in a mouse infection model. Our findings revealed that geraniol effectively inhibits biofilm formation in vitro. Furthermore, in combination with vancomycin, geraniol can reduce the biofilm adhesion to the implant in mice. This suggests the potential of geraniol as an anti-MRSA biofilm drug and can provide a solution for the clinical treatment of biofilm infection.

RevDate: 2022-09-23

Liu Y, Su S, Yu M, et al (2022)

Pyrancoumarin derivative LP4C targeting of pyrimidine de novo synthesis pathway inhibits MRSA biofilm and virulence.

Frontiers in pharmacology, 13:959736 pii:959736.

Staphylococcus aureus poses a serious public health threat because of its multidrug resistance and biofilm formation ability. Hence, developing novel anti-biofilm agents and finding targets are needed to mitigate the proliferation of drug-resistant pathogens. In our previous study, we showed that the pyrancoumarin derivative 2-amino-4-(2,6-dichlorophenyl)-3-cyano-5-oxo-4H, 5H- pyrano [3,2c] chromene (LP4C) can destroy the biofilm of methicillin-resistant S. aureus (MRSA) in vitro and in vivo. Here, we further explored the possible mechanism of LP4C as a potential anti-biofilm drug. We found that LP4C inhibits the expression of enzymes involved in the de novo pyrimidine pathway and attenuates the virulence of MRSA USA300 strain without affecting the agr or luxS quorum sensing system. The molecular docking results indicated that LP4C forms interactions with the key amino acid residues of pyrR protein, which functions as the important regulator of bacterial pyrimidine synthesis. These findings reveal that pyrancoumarin derivative LP4C inhibits MRSA biofilm formation and targeting pyrimidine de novo synthesis pathway.

RevDate: 2022-09-23

Litt PK, Kakani R, Jadeja R, et al (2020)

Effectiveness of Bacteriophages Against Biofilm-Forming Shiga-Toxigenic Escherichia coli on Leafy Greens and Cucumbers.

PHAGE (New Rochelle, N.Y.), 1(4):213-222.

Background: Shiga-toxigenic Escherichia coli (STEC) have caused several produce-associated outbreaks, making it challenging to control these pathogens. Bacteriophages could serve as effective biocontrol. Materials and Methods: Spinach, lettuce, and cucumbers, inoculated with STEC (O157, O26, O45, O103, O111, O121, O145), were treated with lytic bacteriophages and stored at 4°C for 3 days. Surviving STEC were enumerated and observed under scanning electron microscope (SEM), and data analyzed using one-way analysis of variance (ANOVA) (p < 0.05). Results: Bacteriophage treatments significantly reduced STEC populations, compared with the control (p < 0.05). On spinach and romaine, STEC O26, O45, and O103 were reduced to undetectable levels and STEC O157, O111, O121, and O145 by ∼2 logs CFU/cm2. Multiserotype phage cocktail reduced STEC on leafy greens by 1.4 CFU/cm2 and on cucumbers by 1.7 logs CFU/cucumber. Clusters of STEC cells, surrounded by extracellular matrix, were observed under SEM of positive control, whereas phage-treated produce surface showed fewer cells, with cellular damage. Conclusions: Bacteriophages could be utilized as biocontrol against STEC on fresh produce.

RevDate: 2022-09-23

Sarfraz S, Mäntynen PH, Laurila M, et al (2022)

Comparison of Titanium and PEEK Medical Plastic Implant Materials for Their Bacterial Biofilm Formation Properties.

Polymers, 14(18): pii:polym14183862.

This study investigated two of the most commonly used CAD-CAM materials for patient-specific reconstruction in craniomaxillofacial surgery. The aim of this study was to access the biofilm formation of Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis, and Escherichia coli on titanium and PEEK medical implant materials. Two titanium specimens (titanium grade 2 tooled with a Planmeca CAD-CAM milling device and titanium grade 5 tooled with a computer-aided design direct metal laser sintering device (CAD-DMLS)) and one PEEK specimen tooled with a Planmeca CAD-CAM milling device were studied. Bacterial adhesion on implants was evaluated in two groups (saliva-treated group and non-saliva-treated group) to imitate intraoral and extraoral surgical routes for implant placement. The PEEK medical implant material showed higher bacterial adhesion by S. aureus, S. mutans, and E. coli than titanium grade 2 and titanium grade 5, whereas E. faecalis showed higher adhesion to titanium as compared to PEEK. Saliva contamination of implants also effected bacterial attachment. Salivary coating enhanced biofilm formation by S. aureus, S. mutans, and E. faecalis. In conclusion, our findings imply that regardless of the implant material type or tooling techniques used, salivary coating plays a vital role in bacterial adhesion. In addition, the majority of the bacterial strains showed higher adhesion to PEEK than titanium.

RevDate: 2022-09-23

Roy PK, Park SH, Song MG, et al (2022)

Antimicrobial Efficacy of Quercetin against Vibrio parahaemolyticus Biofilm on Food Surfaces and Downregulation of Virulence Genes.

Polymers, 14(18): pii:polym14183847.

For the seafood industry, Vibrio parahaemolyticus, one of the most prevalent food-borne pathogenic bacteria that forms biofilms, is a constant cause of concern. There are numerous techniques used throughout the food supply chain to manage biofilms, but none are entirely effective. Through assessing its antioxidant and antibacterial properties, quercetin will be evaluated for its ability to prevent the growth of V. parahaemolyticus biofilm on shrimp and crab shell surfaces. With a minimum inhibitory concentration (MIC) of 220 µg/mL, the tested quercetin exhibited the lowest bactericidal action without visible growth of bacteria. In contrast, during various experiments in this work, the inhibitory efficacy of quercetin without (control) and with sub-MICs levels (1/2, 1/4, and 1/8 MIC) against V. parahaemolyticus was examined. With increasing quercetin concentration, swarming and swimming motility, biofilm formation, and expression levels of related genes linked to flagella motility (flaA and flgL), biofilm formation (vp0952 and vp0962), and quorum-sensing (luxS and aphA) were all dramatically reduced (p < 0.05). Quercetin (0-110 μg/mL) was investigated on shrimp and crab shell surfaces, the inhibitory effects were 0.68-3.70 and 0.74-3.09 log CFU/cm2, respectively (p < 0.05). The findings were verified using field emission scanning electron microscopy (FE-SEM), which revealed quercetin prevented the development of biofilms by severing cell-to-cell contacts and induced cell lysis, which resulted in the loss of normal cell shape. Furthermore, there was a substantial difference in motility between the treatment and control groups (swimming and swarming). According to our findings, plant-derived quercetin should be used as an antimicrobial agent in the food industry to inhibit the establishment of V. parahaemolyticus biofilms. These findings suggest that bacterial targets are of interest for biofilm reduction with alternative natural food agents in the seafood sector along the entire food production chain.

RevDate: 2022-09-23

Voit M, Trampuz A, M Gonzalez Moreno (2022)

In Vitro Evaluation of Five Newly Isolated Bacteriophages against E. faecalis Biofilm for Their Potential Use against Post-Treatment Apical Periodontitis.

Pharmaceutics, 14(9): pii:pharmaceutics14091779.

State-of-the-art treatment of root canal infection includes the use of mechanical debridement and chemical agents. This disinfection method is limited, and microorganisms can remain in the canal system. Enterococcus faecalis appears with a high prevalence in secondary and persistent root canal infections and can be linked to endodontic treatment failure due to its various resistance mechanisms. Here, we evaluated the activity of newly isolated bacteriophages against clinical isolates of E. faecalis (including one vancomycin- and gentamicin-resistant strain) as a single treatment or in combination with gentamicin and vancomycin. For the resistant strain, daptomycin and fosfomycin were tested. Sixteen E. faecalis strains were used to screen for the presence of bacteriophages in sewage. Five different bacteriophages were characterized in terms of virion morphology, host range and killing-kinetics against each E. faecalis host strain. To investigate the antibiofilm effect of antibiotic and phages, E. faecalis biofilm was grown on porous glass beads and treated with different antibiotic concentrations and with isolated bacteriophages alone or in staggered combinations. A strong biofilm reduction was observed when phages were combined with antibiotic, where combinations with gentamicin showed a better outcome compared to vancomycin. Regarding the resistant strain, daptomycin had a superior antibiofilm effect than fosfomycin.

RevDate: 2022-09-23

Behzadi P, Gajdács M, Pallós P, et al (2022)

Relationship between Biofilm-Formation, Phenotypic Virulence Factors and Antibiotic Resistance in Environmental Pseudomonas&nbsp;aeruginosa.

Pathogens (Basel, Switzerland), 11(9): pii:pathogens11091015.

The formation of a protective biofilm by Pseudomonas aeruginosa (PA) is one of the hallmarks of their survival both in vivo and in harsh environmental conditions, thus, biofilm-eradication has relevance from therapeutic perspectives and for infection control. The aim of our study was to investigate the possible relationship between antibiotic resistance, biofilm-forming capacity and virulence factors in n = 166 PA isolates of environmental origin. Antimicrobial susceptibility testing and the phenotypic detection of resistance determinants were carried out using standard protocols. The biofilm-forming capacity of PA was tested using a standardized crystal violet microtiter plate-based method. Motility (swimming, swarming, and twitching) and siderophore production of the isolates were also assessed. Resistance rates were highest for ciprofloxacin (46.98%), levofloxacin (45.18%), ceftazidime (31.92%) and cefepime (30.12%); 19.28% of isolates met the criteria to be classified as multidrug-resistant (MDR). Efflux pump overexpression, AmpC overexpression, and modified Hodge-test positivity were noted in 28.31%, 18.07% and 3.61%, respectively. 22.89% of isolates were weak/non-biofilm producers, while 27.71% and 49.40% were moderate and strong biofilm producers, respectively. Based on MDR status of the isolates, no significant differences in biofilm-production were shown among environmental PA (non-MDR OD570 [mean ± SD]: 0.416 ± 0.167 vs. MDR OD570: 0.399 ± 0.192; p > 0.05). No significant association was observed between either motility types in the context of drug resistance or biofilm-forming capacity (p > 0.05). 83.13% of isolates tested were positive for siderophore production. The importance of PA as a pathogen in chronic and healthcare-associated infections has been described extensively, while there is increasing awareness of PA as an environmental agent in agriculture and aquaculture. Additional studies in this field would be an important undertaking to understand the interrelated nature of biofilm production and antimicrobial resistance, as these insights may become relevant bases for developing novel therapeutics and eradication strategies against PA.

RevDate: 2022-09-23

Rahman MA, Sahoo N, V Yemmireddy (2022)

Analysis of Sanitizer Rotation on the Susceptibility, Biofilm Forming Ability and Caco-2 Cell Adhesion and Invasion of Listeria.

Pathogens (Basel, Switzerland), 11(9): pii:pathogens11090961.

The purpose of this study was to determine the effect of sanitizer use conditions on the susceptibility, biofilm forming ability and pathogenicity of Listeria&nbsp;monocytogenes. Two different strains of L. monocytogenes and a non-pathogenic L. innocua were exposed to sodium hypochlorite, benzalkonium chloride and peroxyacetic acid at different concentrations (4 to 512 ppm) and treatment times (30 s to 5 min), respectively. Under the tested conditions, no significant difference (p > 0.05) in reduction was observed among the three tested sanitizers. A reduction of 1 to 8 log CFU/mL was observed depending upon the sanitizer concentration and treatment times. The survived cells at the highest sublethal concentration and treatment time of a particular sanitizer upon re-exposure to the same or different sanitizer showed either no change or increased susceptibility when compared to parent strains. Upon repeated exposure to sanitizers at progressively increasing concentrations from 1 to 128 ppm, L. innocua was able to survive concentrations of up to 32 ppm benzalkonium chloride and 64 ppm peroxyacetic acid treatments, respectively. At the tested sub-lethal concentrations, no significant difference (p > 0.05) in biofilm formation was observed among the tested strains. Caco-2 interaction with L. innocua showed a reduction in invasion ability with sublethal concentrations of sanitizers.

RevDate: 2022-09-23

van der Mee-Marquet N, Dos Santos S, Diene SM, et al (2022)

Strong Biofilm Formation and Low Cloxacillin Susceptibility in Biofilm-Growing CC398 Staphylococcus aureus Responsible for Bacteremia in French Intensive Care Units, 2021.

Microorganisms, 10(9): pii:microorganisms10091857.

A prospective 3-month study carried out in 267 ICUs revealed an S. aureus nosocomial bacteremia in one admitted patient out of 110 in adult and pediatric sectors, and in one out of 230 newborns; 242 S. aureus bacteremias occurred during the study, including 7.9% MRSA-bacteremias. In one ICU out of ten, the molecular characteristics, antimicrobial susceptibility profiles and biofilm production of the strains responsible for S. aureus bacteremia were studied. Of the 53 strains studied, 9.4% were MRSA and 52.8% were resistant to erythromycin. MLST showed the predominance of CC398 (37.7% of the strains) followed by CC8 (17.0%), CC45 (13.2%) and CC30 (9.4%). The lukF/S genes were absent from our isolates and tst-1 was found in 9.4% of the strains. Under static conditions and without exposure to glucose, biofilm production was rare (9.4% of the strains, without any CC398). The percentage increased to 62.3% for strains grown in broth supplemented with 1% glucose (including 7 out of 9 CC8 and 17 out of the 20 CC398). Further study of the CC398, including whole genome sequencing, revealed (1) highly frequent patient death within seven days after CC398 bacteremia diagnosis (47.4%), (2) 95.0% of the strains producing biofilm when exposed to sub-inhibitory concentrations of cloxacillin, (3) a stronger biofilm production following exposure to cloxacillin than that observed in broth supplemented with glucose only (p < 0.001), (4) a high minimum biofilm eradication concentration of cloxacillin (128 mg/L) indicating a low cloxacillin susceptibility of biofilm-growing CC398, (5) 95.0% of the strains carrying a ϕSa-3 like prophage and its particular evasion cluster (i.e., yielding chp and scin genes), and (6) 30.0% of the strains carrying a ϕMR11-like prophage and yielding a higher ability to produce biofilm. Our results provide evidence that active surveillance is required to avoid spreading of this virulent staphylococcal clone.

RevDate: 2022-09-23

Shokeen B, Pham E, Esfandi J, et al (2022)

Effect of Calcium Ion Supplementation on Oral Microbial Composition and Biofilm Formation In Vitro.

Microorganisms, 10(9): pii:microorganisms10091780.

The oral cavity contains a variety of ecological niches with very different environmental conditions that shape biofilm structure and composition. The space between the periodontal tissue and the tooth surface supports a unique anaerobic microenvironment that is bathed in the nutrient-rich gingival crevicular fluid (GCF). During the development of periodontitis, this environment changes and clinical findings reported a sustained level of calcium ion concentration in the GCF collected from the periodontal pockets of periodontitis patients. Here, we report the effect of calcium ion supplementation on human oral microbial biofilm formation and community composition employing an established SHI medium-based in vitro model system. Saliva-derived human microbial biofilms cultured in calcium-supplemented SHI medium (SHICa) exhibited a significant dose-dependent increase in biomass and metabolic activity. The effect of SHICa medium on the microbial community composition was evaluated by 16S rRNA gene sequencing using saliva-derived microbial biofilms from healthy donors and periodontitis subjects. In this study, intracellular microbial genomic DNA (iDNA) and extracellular DNA (eDNA) were analyzed separately at the genus level. Calcium supplementation of SHI medium had a differential impact on iDNA and eDNA in the biofilms derived from healthy individuals compared to those from periodontitis subjects. In particular, the genus-level composition of the eDNA portion was distinct between the different biofilms. This study demonstrated the effect of calcium in a unique microenvironment on oral microbial complex supporting the dynamic transformation and biofilm formation.

RevDate: 2022-09-23

Henrici De Angelis L, Stirpe M, Tomolillo D, et al (2022)

The Multifunctional Role of Poloxamer P338 as a Biofilm Disrupter and Antibiotic Enhancer: A Small Step forward against the Big Trouble of Catheter-Associated Escherichia coli Urinary Tract Infections.

Microorganisms, 10(9): pii:microorganisms10091757.

Poloxamer 338 (P338), a nonionic surfactant amphiphilic copolymer, is herein proposed as an anti-biofilm compound for the management of catheter-associated urinary tract infections (CAUTIs). P338's ability to disrupt Escherichia coli biofilms on silicone urinary catheters and to serve as antibiotic enhancer was evaluated for biofilm-producing E. coli Ec5FSL and Ec9FSL clinical strains, isolated from urinary catheters. In static conditions, quantitative biofilm formation assay allowed us to determine the active P338 concentration. In dynamic conditions, the BioFlux system, combined with confocal laser scanning microscopy, allowed us to investigate the P338 solution's ability to detach biofilm, alone or in combination with sub-MIC concentrations of cefoxitin (FOX). The 0.5% P338 solution was able to destroy the structure of E. coli biofilms, to reduce the volume and area fraction covered by adherent cells (41.42 ± 4.79% and 56.20 ± 9.22% reduction for the Ec5FSL and Ec9FSL biofilms, respectively), and to potentiate the activity of 1\2 MIC FOX in disaggregating biofilms (19.41 ± 7.41% and 34.66 ± 3.75% reduction in the area fraction covered by biofilm for Ec5FSL and Ec9FSL, respectively) and killing cells (36.85 ± 7.13% and 32.33 ± 4.65% increase in the biofilm area covered by dead Ec5FSL and Ec9FSL cells, respectively).

RevDate: 2022-09-23

Tan HC, Cheung GSP, Chang JWW, et al (2022)

Enterococcus faecalis Shields Porphyromonas gingivalis in Dual-Species Biofilm in Oxic Condition.

Microorganisms, 10(9): pii:microorganisms10091729.

AIM: To develop a reproducible biofilm model consisting of Enterococcus faecalis (E. faecalis) and Porphyromonas gingivalis (P. gingivalis) and to evaluate the interaction between the two bacterial species.

METHODOLOGY: E. faecalis and P. gingivalis were grown in mono-culture, sequential, and co-culture models for 96 h in a 96-well polystyrene microtiter plate under both aerobic and anaerobic conditions separately. The viability of the two bacterial species in the biofilms was quantified by polymerase chain reaction (qPCR). Biofilm thickness and protein contents were measured using confocal laser scanning microscopy (CLSM). Two-way analysis of variance (ANOVA) was performed to analyze cell viability and biofilm thickness among different culture models cultivated under either aerobic or anaerobic conditions. The level of significance was set at p < 0.05.

RESULTS: Different culture models tested did not show any significant difference between the viable cell counts of both E. faecalis and P. gingivalis cultivated under aerobic and anaerobic conditions (p > 0.05). Biofilm was significantly thicker (p < 0.05) in the co-culture models compared to the mono-culture and sequential models. Protein contents in the biofilms were more pronounced when both bacterial species were co-cultured under aerobic conditions.

CONCLUSIONS: E. faecalis appeared to shield P. gingivalis and support its continued growth in oxic (aerobic) conditions. The co-culture model of E. faecalis and P. gingivalis produced a significantly thicker biofilm irrespective of the presence or absence of oxygen, while increased protein contents were only observed in the presence of oxygen.

RevDate: 2022-09-23

Gumber HK, Louyakis AS, Sarma T, et al (2022)

Effect of a Stannous Fluoride Dentifrice on Biofilm Composition, Gene Expression and Biomechanical Properties.

Microorganisms, 10(9): pii:microorganisms10091691.

An in situ study was conducted to examine the mode of action of a 0.454% stannous fluoride (SnF2)-containing dentifrice in controlling the composition and properties of oral biofilm. Thirteen generally healthy individuals participated in the study. Each participant wore an intra-oral appliance over a 48-h period to measure differences in the resulting biofilm's architecture, mechanical properties, and bacterial composition after using two different toothpaste products. In addition, metatranscriptomics analysis of supragingival plaque was conducted to identify the gene pathways influenced. The thickness and volume of the microcolonies formed when brushing with the SnF2 dentifrice were dramatically reduced compared to the control 0.76% sodium monofluorophosphate (MFP)-containing toothpaste. Similarly, the biophysical and nanomechanical properties measured by atomic force microscopy (AFM) demonstrated a significant reduction in biofilm adhesive properties. Metatranscriptomic analysis identified pathways associated with biofilm formation, cell adhesion, quorum sensing, and N-glycosylation that are significantly downregulated with SnF2. This study provides a clinically relevant snapshot of how the use of a stabilized, SnF2 toothpaste formulation can change the spatial organization, nanomechanical, and gene expression properties of bacterial communities.

RevDate: 2022-09-23

Grzegorczyk M, Pogorzelski S, Janowicz P, et al (2022)

Micron-Scale Biogeography of Seawater Biofilm Colonies at Submersed Solid Substrata Affected by Organic Matter and Microbiome Transformation in the Baltic Sea.

Materials (Basel, Switzerland), 15(18): pii:ma15186351.

The aim of this research was to determine temporal and spatial evolution of biofilm architecture formed at model solid substrata submersed in Baltic sea coastal waters in relation to organic matter transformation along a one-year period. Several materials (metals, glass, plastics) were deployed for a certain time, and the collected biofilm-covered samples were studied with a confocal microscopy technique using the advanced programs of image analysis. The geometric and structural biofilm characteristics: biovolume, coverage fraction, mean thickness, spatial heterogeneity, roughness, aggregation coefficient, etc., turned out to evolve in relation to organic matter transformation trends, trophic water status, microbiome evolution, and biofilm micro-colony transition from the heterotrophic community (mostly bacteria) to autotrophic (diatom-dominated) systems. The biofilm morphology parameters allowed the substratum roughness, surface wettability, chromatic organisms colony adaptation to substrata, and quorum sensing or cell to cell signaling effects to be quantitatively evaluated. In addition to the previous work, the structural biofilm parameters could become further novel trophic state indicators.

RevDate: 2022-09-23

Pouget C, Dunyach-Remy C, Magnan C, et al (2022)

Polymicrobial Biofilm Organization of Staphylococcus aureus and Pseudomonas aeruginosa in a Chronic Wound Environment.

International journal of molecular sciences, 23(18): pii:ijms231810761.

Biofilm on the skin surface of chronic wounds is an important step that involves difficulties in wound healing. The polymicrobial nature inside this pathogenic biofilm is key to understanding the chronicity of the lesion. Few in vitro models have been developed to study bacterial interactions inside this chronic wound. We evaluated the biofilm formation and the evolution of bacteria released from this biofilm on the two main bacteria isolated in this condition, Staphylococcus aureus and Pseudomonas aeruginosa, using a dynamic system (BioFlux™ 200) and a chronic wound-like medium (CWM) that mimics the chronic wound environment. We observed that all species constituted a faster biofilm in the CWM compared to a traditional culture medium (p < 0.01). The percentages of biofilm formation were significantly higher in the mixed biofilm compared to those determined for the bacterial species alone (p < 0.01). Biofilm organization was a non-random structure where S. aureus aggregates were located close to the wound surface, whereas P. aeruginosa was located deeper in the wound bed. Planktonic biofilm-detached bacteria showed decreased growth, overexpression of genes encoding biofilm formation, and an increase in the mature biofilm biomass formed. Our data confirmed the impact of the chronic wound environment on biofilm formation and on bacterial lifecycle inside the biofilm.

RevDate: 2022-09-23

Kim S, Lee JH, Kim YG, et al (2022)

Hydroquinones Inhibit Biofilm Formation and Virulence Factor Production in Staphylococcus aureus.

International journal of molecular sciences, 23(18): pii:ijms231810683.

Staphylococcus aureus is one of the major pathogens responsible for antimicrobial resistance-associated death. S. aureus can secrete various exotoxins, and staphylococcal biofilms play critical roles in antibiotic tolerance and the persistence of chronic infections. Here, we investigated the inhibitory effects of 18 hydroquinones on biofilm formation and virulence factor production by S. aureus. It was found that 2,5-bis(1,1,3,3-tetramethylbutyl) hydroquinone (TBHQ) at 1 µg/mL efficiently inhibits biofilm formation by two methicillin-sensitive and two methicillin-resistant S. aureus strains with MICs of 5 µg/mL, whereas the backbone compound hydroquinone did not (MIC > 400 µg/mL). In addition, 2,3-dimethylhydroquinone and tert-butylhydroquinone at 50 µg/mL also exhibited antibiofilm activity. TBHQ at 1 µg/mL significantly decreased the hemolytic effect and lipase production by S. aureus, and at 5-50 µg/mL was non-toxic to the nematode Caenorhabditis elegans and did not adversely affect Brassica rapa seed germination or growth. Transcriptional analyses showed that TBHQ suppressed the expression of RNAIII (effector of quorum sensing). These results suggest that hydroquinones, particularly TBHQ, are potentially useful for inhibiting S. aureus biofilm formation and virulence.

RevDate: 2022-09-23

Klagisa R, Racenis K, Broks R, et al (2022)

Analysis of Microorganism Colonization, Biofilm Production, and Antibacterial Susceptibility in Recurrent Tonsillitis and Peritonsillar Abscess Patients.

International journal of molecular sciences, 23(18): pii:ijms231810273.

BACKGROUND: Despite the widespread use of antibiotics to treat infected tonsils, episodes of tonsillitis tend to recur and turn into recurrent tonsillitis (RT) or are complicated by peritonsillar abscesses (PTAs). The treatment of RT and PTAs remains surgical, and tonsillectomies are still relevant.

MATERIALS AND METHODS: In a prospective, controlled study, we analyzed the bacteria of the tonsillar crypts of 99 patients with RT and 29 patients with a PTA. We performed the biofilm formation and antibacterial susceptibility testing of strains isolated from study patients. We compared the results obtained between patient groups with the aim to identify any differences that may contribute to ongoing symptoms of RT or that may play a role in developing PTAs.

RESULTS: The greatest diversity of microorganisms was found in patients with RT. Gram-positive bacteria were predominant in both groups. Candida species were predominant in patients with a PTA (48.3% of cases). Irrespective of patient group, the most commonly isolated pathogenic bacterium was S. aureus (in 33.3% of RT cases and in 24.14% of PTA cases). The most prevalent Gram-negative bacterium was K. pneumoniae (in 10.1% of RT cases and in 13.4% of PTA cases). At least one biofilm-producing strain was found in 37.4% of RT cases and in 27.6% of PTA cases. Moderate or strong biofilm producers were detected in 16 out of 37 cases of RT and in 2 out of 8 PTA cases. There was a statistically significant association found between the presence of Gram-positive bacteria and a biofilm-formation phenotype in the RT group and PTA group (Pearson χ2 test, p < 0.001). S. aureus and K. pneumoniae strains were sensitive to commonly used antibiotics. One S. aureus isolate was identified as MRSA.

CONCLUSIONS: S. aureus is the most common pathogen isolated from patients with RT, and Candida spp. are the most common pathogens isolated from patients with a PTA. S. aureus isolates are susceptible to most antibiotics. Patients with RT more commonly have biofilm-producing strains, but patients with a PTA more commonly have biofilm non-producer strains. K. pneumoniae does not play a major role in biofilm production.

RevDate: 2022-09-23

Masebe RD, MS Thantsha (2022)

Anti-Biofilm Activity of Cell Free Supernatants of Selected Lactic Acid Bacteria against Listeria monocytogenes Isolated from Avocado and Cucumber Fruits, and from an Avocado Processing Plant.

Foods (Basel, Switzerland), 11(18): pii:foods11182872.

Listeria monocytogenes forms biofilms on food contact surfaces, a niche from where it dislodges to contaminate food products including fresh produce. Probiotics and their derivatives are considered promising alternative strategies to curb the presence of L. monocytogenes in varied food applications. Nonetheless, studies on their anti-biofilm effects against L. monocytogenes from avocados and cucumbers are sparse. This study screened the biofilm formation capabilities of L. monocytogenes strains Avo and Cuc isolated from the avocado and cucumber fruits respectively, and strain 243 isolated from an avocado processing plant; and evaluated the anti-biofilm effects of cell free supernatants (CFS) of Lactobacillus acidophilus La14 150B, Lactiplantibacillus plantarum B411 and Lacticaseibacillus rhamnosus ATCC 53103 against their biofilms formed on polyvinyl chloride (PVC) and stainless steel. All the L. monocytogenes strains formed biofilms (classified either as moderate or strong biofilm formers) on these materials. The presence of CFS reduced the biofilm formation capabilities of these strains and disrupted the integrity of their pre-formed biofilms. Quantitative reverse transcriptase polymerase chain reaction revealed significant reduction of positive regulatory factor A (prfA) gene expression by L. monocytogenes biofilm cells in the presence of CFS (p < 0.05). Thus, these CFS have potential as food grade sanitizers for control of L. monocytogenes biofilms in the avocado and cucumber processing facilities.

RevDate: 2022-09-23

Ejaz H, Junaid K, Yasmeen H, et al (2022)

Multiple Antimicrobial Resistance and Heavy Metal Tolerance of Biofilm-Producing Bacteria Isolated from Dairy and Non-Dairy Food Products.

Foods (Basel, Switzerland), 11(18): pii:foods11182728.

Foodborne pathogens have acquired the ability to produce biofilms to survive in hostile environments. This study evaluated biofilm formation, antimicrobial resistance (AMR), and heavy metal tolerance of bacteria isolated from dairy and non-dairy food products. We aseptically collected and processed 200 dairy and non-dairy food specimens in peptone broth, incubated them overnight at 37 °C, and sub-cultured them on various culture media. Bacterial growth was identified with biochemical tests and API 20E and 20NE strips. The AMR of the isolates was observed against different antibacterial drug classes. Biofilm formation was detected with the crystal violet tube method. Heavy metal salts were used at concentrations of 250-1500 µg/100 mL to observe heavy metal tolerance. We isolated 180 (50.4%) bacteria from dairy and 177 (49.6%) from non-dairy food samples. The average colony-forming unit (CFU) count for dairy and non-dairy samples was 2.9 ± 0.9 log CFU/mL and 5.1 ± 0.3 log CFU/mL, respectively. Corynebacterium kutscheri (n = 74), lactobacilli (n = 73), and Staphylococcus aureus (n = 56) were the predominant Gram-positive and Shigella (n = 10) the predominant Gram-negative bacteria isolated. The correlation between biofilm formation and AMR was significant (p < 0.05) for most cephalosporins, aminoglycosides, and fluoroquinolones. Heavy metal tolerance tended to be higher in biofilm producers at different metal concentrations. The pathogens isolated from dairy and non-dairy food showed a high burden of AMR, high propensity for biofilm formation, and heavy metal tolerance, and pose an imminent threat to public health.

RevDate: 2022-09-23

Thomsen K, Høiby N, Jensen PØ, et al (2022)

Immune Response to Biofilm Growing Pulmonary Pseudomonas aeruginosa Infection.

Biomedicines, 10(9): pii:biomedicines10092064.

Biofilm infections are tolerant to the host responses and recalcitrance to antibiotic drugs and disinfectants. The induced host-specific innate and adaptive immune responses by established biofilms are significantly implicated and contributes to the course of the infections. Essentially, the host response may be the single one factor impacting the outcome most, especially in cases where the biofilm is caused by low virulent opportunistic bacterial species. Due to the chronicity of biofilm infections, activation of the adaptive immune response mechanisms is frequently experienced, and instead of clearing the infection, the adaptive response adds to the pathogenesis. To a high degree, this has been reported for chronic Pseudomonas aeruginosa lung infections, where both a pronounced antibody response and a skewed Th1/Th2 balance has been related to a poorer outcome. In addition, detection of an adaptive immune response can be used as a significant indicator of a chronic P. aeruginosa lung infection and is included in the clinical definitions as such. Those issues are presented in the present review, along with a characterization of the airway structure in relation to immune responses towards P. aeruginosa pulmonary infections.

RevDate: 2022-09-23

Jubair N, R M, Fatima A, et al (2022)

Evaluation of Catechin Synergistic and Antibacterial Efficacy on Biofilm Formation and acrA Gene Expression of Uropathogenic E. coli Clinical Isolates.

Antibiotics (Basel, Switzerland), 11(9): pii:antibiotics11091223.

Uropathogenic Escherichia coli has a propensity to build biofilms to resist host defense and antimicrobials. Recurrent urinary tract infection (UTI) caused by multidrug-resistant, biofilm-forming E. coli is a significant public health problem. Consequently, searching for alternative medications has become essential. This study was undertaken to investigate the antibacterial, synergistic, and antibiofilm activities of catechin isolated from Canarium patentinervium Miq. against three E. coli ATCC reference strains (ATCC 25922, ATCC 8739, and ATCC 43895) and fifteen clinical isolates collected from UTI patients in Baghdad, Iraq. In addition, the expression of the biofilm-related gene, acrA, was evaluated with and without catechin treatment. Molecular docking was performed to evaluate the binding mode between catechin and the target protein using Autodock Vina 1.2.0 software. Catechin demonstrated significant bactericidal activity with a minimum inhibitory concentration (MIC) range of 1-2 mg/mL and a minimum bactericidal concentration (MBC) range of 2-4 mg/mL and strong synergy when combined with tetracycline at the MBC value. In addition, catechin substantially reduced E. coli biofilm by downregulating the acrA gene with a reduction percent ≥ 60%. In silico analysis revealed that catechin bound with high affinity (∆G = -8.2 kcal/mol) to AcrB protein (PDB-ID: 5ENT), one of the key AcrAB-TolC efflux pump proteins suggesting that catechin might inhibit the acrA gene indirectly by docking at the active site of AcrB protein.

RevDate: 2022-09-23

Kunz Coyne AJ, Stamper K, Kebriaei R, et al (2022)

Phage Cocktails with Daptomycin and Ampicillin Eradicates Biofilm-Embedded Multidrug-Resistant Enterococcus faecium with Preserved Phage Susceptibility.

Antibiotics (Basel, Switzerland), 11(9): pii:antibiotics11091175.

Multidrug-resistant (MDR) Enterococcus faecium is a challenging nosocomial pathogen known to colonize medical device surfaces and form biofilms. Bacterio (phages) may constitute an emerging anti-infective option for refractory, biofilm-mediated infections. This study evaluates eight MDR E. faecium strains for biofilm production and phage susceptibility against nine phages. Two E. faecium strains isolated from patients with bacteremia and identified to be biofilm producers, R497 (daptomycin (DAP)-resistant) and HOU503 (DAP-susceptible dose-dependent (SDD), in addition to four phages with the broadest host ranges (ATCC 113, NV-497, NV-503-01, NV-503-02) were selected for further experiments. Preliminary phage-antibiotic screening was performed with modified checkerboard minimum biofilm inhibitory concentration (MBIC) assays to efficiently screen for bacterial killing and phage-antibiotic synergy (PAS). Data were compared by one-way ANOVA and Tukey (HSD) tests. Time kill analyses (TKA) were performed against R497 and HOU503 with DAP at 0.5× MBIC, ampicillin (AMP) at free peak = 72 µg/mL, and phage at a multiplicity of infection (MOI) of 0.01. In 24 h TKA against R497, phage-antibiotic combinations (PAC) with DAP, AMP, or DAP + AMP combined with 3- or 4-phage cocktails demonstrated significant killing compared to the most effective double combination (ANOVA range of mean differences 2.998 to 3.102 log10 colony forming units (CFU)/mL; p = 0.011, 2.548 to 2.868 log10 colony forming units (CFU)/mL; p = 0.023, and 2.006 to 2.329 log10 colony forming units (CFU)/mL; p = 0.039, respectively), with preserved phage susceptibility identified in regimens with 3-phage cocktails containing NV-497 and the 4-phage cocktail. Against HOU503, AMP combined with any 3- or 4-phage cocktail and DAP + AMP combined with the 3-phage cocktail ATCC 113 + NV-497 + NV-503-01 demonstrated significant PAS and bactericidal activity (ANOVA range of mean differences 2.251 to 2.466 log10 colony forming units (CFU)/mL; p = 0.044 and 2.119 to 2.350 log10 colony forming units (CFU)/mL; p = 0.028, respectively), however, only PAC with DAP + AMP maintained phage susceptibility at the end of 24 h TKA. R497 and HOU503 exposure to DAP, AMP, or DAP + AMP in the presence of single phage or phage cocktail resulted in antibiotic resistance stabilization (i.e., no antibiotic MBIC elevation compared to baseline) without identified antibiotic MBIC reversion (i.e., lowering of antibiotic MBIC compared to baseline in DAP-resistant and DAP-SDD isolates) at the end of 24 h TKA. In conclusion, against DAP-resistant R497 and DAP-SDD HOU503 E. faecium clinical blood isolates, the use of DAP + AMP combined with 3- and 4-phage cocktails effectively eradicated biofilm-embedded MDR E. faecium without altering antibiotic MBIC or phage susceptibility compared to baseline.

RevDate: 2022-09-23

Roy PK, Song MG, SY Park (2022)

The Inhibitory Effect of Quercetin on Biofilm Formation of Listeria monocytogenes Mixed Culture and Repression of Virulence.

Antioxidants (Basel, Switzerland), 11(9): pii:antiox11091733.

Listeria monocytogenes is the species of foodborne pathogenic bacteria that causes the infection listeriosis. The food production chain employs various methods to control biofilms, although none are completely successful. This study evaluates the effectiveness of quercetin as a food additive in reducing L. monocytogenes mixed cultures (ATCC19113, ATCC19117, and ATCC15313) biofilm formation on stainless steel (SS), silicon rubber (SR), and hand glove (HG) coupons, as well as tests its antimicrobial activities. With a minimum inhibitory concentration (MIC) of 250 µg/mL, the tested quercetin exhibited the lowest bactericidal action with no visible bacterial growth. In contrast, during various experiments in this work, the inhibitory efficacy of quercetin at sub-MICs levels (1/2, 1/4, and 1/8 MIC) against L. monocytogenes was examined. A control group was not added with quercetin. The current study also investigates the effect of quercetin on the expression of different genes engaged in motility (flaA, fbp), QS (agrA), and virulence (hlyA, prfA). Through increasing quercetin concentration, swarming and swimming motility, biofilm formation, and expression levels of target genes linked to flagella motility, virulence, and quorum-sensing were all dramatically reduced. Quercetin (0-125 μg/mL) was investigated on the SS, SR, and HG surfaces; the inhibitory effects were 0.39-2.07, 0.09-1.96 and 0.03-1.69 log CFU/cm2, respectively (p < 0.05). Field-emission scanning electron microscopy (FE-SEM) corroborated the findings because quercetin prevented the development of biofilms by severing cell-to-cell contacts and inducing cell lysis, which resulted in the loss of normal cell shape. Our findings suggest that plant-derived quercetin should be used as an antimicrobial agent in the food industry to control the development of L. monocytogenes biofilms. These outcomes suggest that bacterial targets are of interest for biofilm reduction, with alternative natural food agents in the food sector along the entire food production chain.

RevDate: 2022-09-23

Dolma KG, Khati R, Paul AK, et al (2022)

Virulence Characteristics and Emerging Therapies for Biofilm-Forming Acinetobacter baumannii: A Review.

Biology, 11(9): pii:biology11091343.

Acinetobacter species is one of the most prevailing nosocomial pathogens with a potent ability to develop antimicrobial resistance. It commonly causes infections where there is a prolonged utilization of medical devices such as CSF shunts, catheters, endotracheal tubes, and similar. There are several strains of Acinetobacter (A) species (spp), among which the majority are pathogenic to humans, but A. baumannii are entirely resistant to several clinically available antibiotics. The crucial mechanism that renders them a multidrug-resistant strain is their potent ability to synthesize biofilms. Biofilms provide ample opportunity for the microorganisms to withstand the harsh environment and further cause chronic infections. Several studies have enumerated multiple physiological and virulence factors responsible for the production and maintenance of biofilms. To further enhance our understanding of this pathogen, in this review, we discuss its taxonomy, pathogenesis, current treatment options, global resistance rates, mechanisms of its resistance against various groups of antimicrobials, and future therapeutics.

RevDate: 2022-09-22

Bullard S, Mona M, Pereira AC, et al (2022)

Quantitative Analysis of Biofilm Removal Following Instrumentation with TRUShape and Vortex Blue File Systems: Microbiological Study.

Frontiers in bioscience (Scholar edition), 14(3):18.

Mesial roots and isthmuses of mandibular molars are difficult areas to obtain adequate disinfection of root canal walls, and consequently microorganisms can survive treatment. The present study compared, through real-time polymerase chain reaction (qPCR), the effectiveness of TRUShape (TS) (Dentsply Tulsa Dental Specialties, Tulsa, OK) and Vortex Blue (VB) (Dentsply Tulsa Dental Specialties, Tulsa, OK) in removing Enterococcus faecalis (E. faecalis) from the mesial canals and isthmuses of mandibular molars. Fifty extracted human lower molars were inoculated with E. faecalis OG1RF for 14 days, and then an initial bacterial sample was collected with paper points from mesiobuccal and mesiolingual canals and isthmuses. The specimens were randomly divided into four groups (n = 10 teeth; 20 canals each), according to instrumentation system: TS 25/0.06, TS 30/0.06, VB 25/0.06 and VB 30/0.06. The remaining 10 teeth were divided between positive control, inoculated teeth without instrumentation or irrigation, and negative controls, teeth without inoculation. After instrumentation, the final sample was taken using paper points and DNA was isolated. Primers specific for E. faecalis were used for qPCR. The bacterial reduction between pre- and post-instrumentation was calculated. One-way analysis of variance (ANOVA) with Bonferroni's multiple-comparisons tests were for statistical analysis with significance of (p < 0.05). All file systems were able to reduce the load of E. faecalis from the prepared root canals, however, TS size 30 removed significantly more bacteria than size 25. Interestingly, regardless of the size, TS files removed significantly more E. faecalis biofilm (p < 0.05) than did VB files (63.7% vs 50.8% for size 25, and 69.5% vs 56% for size 30). In conclusion, when combined with irrigation, TS file system is more effective than VB in reducing E. faecalis biofilms from mesiobuccal and mesiolingual canals and the isthmuses of mandibular molars.

RevDate: 2022-09-23

Dzib-Baak HE, Uc-Cachón AH, Dzul-Beh AJ, et al (2022)

Efficacy of Fosfomycin against Planktonic and Biofilm-Associated MDR Uropathogenic Escherichia coli Clinical Isolates.

Tropical medicine and infectious disease, 7(9): pii:tropicalmed7090235.

Urinary tract infections (UTI) are a severe public health problem and are caused mainly by the uropathogenic Escherichia coli (UPEC). Antimicrobial resistance and limited development of new antimicrobials have led to the reuse of old antibiotics such as fosfomycin. The aim of this study was to evaluate the in vitro efficacy of fosfomycin on a collection of multidrug-resistant (MDR) UPEC and the degradative activity on biofilm producers. A total of 100 MDR UPEC clinical isolates were collected from patients at Mexican second- and third-level hospitals. Microorganism identification was performed using an automated system, the evaluation of the susceptibility of clinical isolates to fosfomycin was performed using the resazurin microtiter assay, and the identification of biofilm producers and the effect of fosfomycin in biofilms were evaluated using the crystal violet method. Among planktonic MDR UPEC, 93% were susceptible to fosfomycin. Eighty-three MDR UPEC were categorized as weak (39.8%), moderate (45.2%), and strong (14.5%) biofilm producers. Fosfomycin exhibited degradative activity ranging from 164.4 µg/mL to 1045 µg/mL. Weak producers required statistically lower concentrations of fosfomycin to destroy the biofilm, contrary to moderate and strong producers. In conclusion, fosfomycin could be an option for the treatment of infections caused by MDR UPEC, for which the antimicrobial treatment is more often becoming limited.

RevDate: 2022-09-22

Madan R, Madan S, A Hussain (2022)

Kinetic Study for Startup of Aerobic Moving Bed Biofilm Reactor in Treatment of Textile Dye Wastewater.

Applied biochemistry and biotechnology [Epub ahead of print].

Due to high augmentation in population and low availability of land, the quantum of wastewater production has surged resulting in advancements in wastewater treatment systems. To cope under such stressful circumstances, moving bed biofilm reactor (MBBR) proves to be an upgraded treatment technology for industrial and municipal wastewater treatment. The present startup study has been carried out using a laboratory-scale aerobic MBBR with working volume of 25L for textile dye wastewater treatment having AnoxKaldnes K3 media at filling percentage of 50%. In order to acclimatize the microorganisms on textile dye wastewater, the startup of the reactor was carried out using lactose as readily degradable co-substrate with textile dye wastewater in different ratios at hydraulic retention time (HRT) of 24 h. The biofilm on the media was developed in 63 days duration and the reactor attained pseudo-steady state (PSS) in 185 days period. During PSS condition of the MBBR, the maximum chemical oxygen demand (COD) removal efficiency of 92% with mixed liquor suspended solids (MLSS) concentration of 4224 ± 22 mg/L has been achieved. The kinetic study for biodegradation of textile dye wastewater has also been carried out using the Monod growth kinetics. The values of bio-kinetic coefficients of yield of heterotrophic biomass (Y) and endogenous decay coefficient for heterotrophic biomass (Kd) recorded are 0.394 mgVSS/mgCOD.d and 0.087 day-1, respectively. The values of specific substrate removal rate (k), Monod half saturation constant (Ks), and maximum specific growth rate for heterotrophic biomass (µmax) are 0.024 mgCOD/mgVSS.d, 53.203 mg/L, and 0.0095 day-1, respectively, demonstrating the suitability and healthy performance of MBBR for textile dye wastewater treatment.

RevDate: 2022-09-23

Sousa IS, Mello TP, Pereira EP, et al (2022)

Biofilm Formation by Chromoblastomycosis Fungi Fonsecaea pedrosoi and Phialophora verrucosa: Involvement with Antifungal Resistance.

Journal of fungi (Basel, Switzerland), 8(9): pii:jof8090963.

Patients with chromoblastomycosis (CBM) suffer chronic tissue lesions that are hard to treat. Considering that biofilm is the main growth lifestyle of several pathogens and it is involved with both virulence and resistance to antimicrobial drugs, we have investigated the ability of CBM fungi to produce this complex, organized and multicellular structure. Fonsecaea pedrosoi and Phialophora verrucosa conidial cells were able to adhere on a polystyrene abiotic substrate, differentiate into hyphae and produce a robust viable biomass containing extracellular matrix. Confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) showed the tridimensional architecture of the mature biofilms, revealing a dense network of interconnected hyphae, inner channels and amorphous extracellular polymeric material. Interestingly, the co-culture of each fungus with THP-1 macrophage cells, used as a biotic substrate, induced the formation of a mycelial trap covering and damaging the macrophages. In addition, the biofilm-forming cells of F. pedrosoi and P. verrucosa were more resistant to the conventional antifungal drugs than the planktonic-growing conidial cells. The efflux pump activities of P. verrucosa and F. pedrosoi biofilms were significantly higher than those measured in conidia. Taken together, the data pointed out the biofilm formation by CBM fungi and brought up a discussion of the relevance of studies about their antifungal resistance mechanisms.

RevDate: 2022-09-23

Park J, Kim H, Kang HK, et al (2022)

Lycosin-II Exhibits Antifungal Activity and Inhibits Dual-Species Biofilm by Candida albicans and Staphylococcus aureus.

Journal of fungi (Basel, Switzerland), 8(9): pii:jof8090901.

The increase and dissemination of antimicrobial resistance is a global public health issue. To address this, new antimicrobial agents have been developed. Antimicrobial peptides (AMPs) exhibit a wide range of antimicrobial activities against pathogens, including bacteria and fungi. Lycosin-II, isolated from the venom of the spider Lycosa singoriensis, has shown antibacterial activity by disrupting membranes. However, the mode of action of Lycosin-II and its antifungal activity have not been clearly described. Therefore, we confirmed that Lycosin-II showed antifungal activity against Candida albicans (C. albicans). To investigate the mode of action, membrane-related assays were performed, including an evaluation of C. albicans membrane depolarization and membrane integrity after exposure to Lycosin-II. Our results indicated that Lycosin-II damaged the C. albicans membrane. Additionally, Lycosin-II induced oxidative stress through the generation of reactive oxygen species (ROS) in C. albicans. Moreover, Lycosin-II exhibited an inhibitory effect on dual-species biofilm formation by C. albicans and Staphylococcus aureus (S. aureus), which are the most co-isolated fungi and bacteria. These results revealed that Lycosin-II can be utilized against C. albicans and dual-species strain infections.

RevDate: 2022-09-22

Tahir S, Emanuel S, Inglis DW, et al (2022)

Mild Positive Pressure Improves the Efficacy of Benzalkonium Chloride against Staphylococcus aureus Biofilm.

Bioengineering (Basel, Switzerland), 9(9): pii:bioengineering9090461.

Current protocols using liquid disinfectants to disinfect heat-sensitive hospital items frequently fail, as evidenced by the continued isolation of bacteria following decontamination. The contamination is, in part, due to biofilm formation. We hypothesize that mild positive pressure (PP) will disrupt this biofilm structure and improve liquid disinfectant/detergent penetration to biofilm bacteria for improved killing. Staphylococcus aureus biofilm, grown on polycarbonate coupons in the biofilm reactor under shear at 35 °C for 3 days, was treated for 10 min and 60 min with various dilutions of benzalkonium chloride without PP at 1 atmosphere (atm), and with PP at 3, 5, 7, and 10 atm. The effect on biofilm and residual bacterial viability was determined by standard plate counts, confocal laser scanning microscopy, and scanning electron microscopy. Combined use of benzalkonium chloride and PP up to 10 atm significantly increased biofilm killing up to 4.27 logs, as compared to the treatment using disinfectant alone. Microscopy results were consistent with the viability plate count results. PP improved disinfectant efficacy against bacterial biofilm. The use of mild PP is possible in many flow situations or if equipment/contaminated surfaces can be placed in a pressure chamber.

RevDate: 2022-09-21

Maharjan S, Ansari M, Maharjan P, et al (2022)

Phenotypic detection of methicillin resistance, biofilm production, and inducible clindamycin resistance in Staphylococcus aureus clinical isolates in Kathmandu, Nepal.

Tropical medicine and health, 50(1):71.

INTRODUCTION: Methicillin resistance, inducible clindamycin resistance (ICR), biofilm production, and increased minimum inhibitory concentration (MIC) of vancomycin in Staphylococcus aureus are major causes of antibiotic treatment failure and increased morbidity and mortality. The surveillance of such isolates and the study of their antimicrobial pattern are essential in managing the infections caused by these isolates. This study aimed to determine methicillin resistance, biofilm production, and ICR in S. aureus isolates from a tertiary care hospital in Kathmandu, Nepal.

MATERIALS AND METHODS: A total of 217 S. aureus isolated from different samples were processed following standard laboratory procedures. Antibiotic susceptibility testing was performed by the Kirby-Bauer disk diffusion technique. Methicillin-resistant S. aureus (MRSA) were identified by the cefoxitin disk diffusion test, and biofilm producers were examined using the microtiter plate technique. D-test and E-test were performed to determine inducible clindamycin resistance and minimum inhibitory concentration of vancomycin, respectively.

RESULTS: Among the 217 S. aureus isolates, 78.3% were multidrug-resistant (MDR), 47.0% were MRSA, 62.2% were biofilm producers, and 50.7% showed ICR. All MRSA isolates exhibited MIC levels of vancomycin within the susceptible range. Biofilm producers and MRSA isolates showed elevated antimicrobial resistance. MRSA was significantly associated with MDR. Biofilm-producing and multidrug-resistant MRSA isolates showed significantly higher MIC levels of vancomycin (p = 0.0013 and < 0.0001, respectively), while ICR was significantly higher in MDR (p = 0.0001) isolates.

CONCLUSION: High multidrug resistance, MRSA, and ICR in this study call for routine evaluation of antibiotic susceptibility patterns of S. aureus. Vancomycin can be used to treat serious staphylococcal infections. Clindamycin should be prescribed only after performing the D-test. Drugs like teicoplanin, chloramphenicol, doxycycline, amikacin, and levofloxacin can treat MRSA infections.

RevDate: 2022-09-21

Létoffé S, Wu Y, Darch SE, et al (2022)

Pseudomonas aeruginosa Production of Hydrogen Cyanide Leads to Airborne Control of Staphylococcus aureus Growth in Biofilm and In Vivo Lung Environments.

mBio [Epub ahead of print].

Diverse bacterial volatile compounds alter bacterial stress responses and physiology, but their contribution to population dynamics in polymicrobial communities is not well known. In this study, we showed that airborne volatile hydrogen cyanide (HCN) produced by a wide range of Pseudomonas aeruginosa clinical strains leads to at-a-distance in vitro inhibition of the growth of a wide array of Staphylococcus aureus strains. We determined that low-oxygen environments not only enhance P. aeruginosa HCN production but also increase S. aureus sensitivity to HCN, which impacts P. aeruginosa-S. aureus competition in microaerobic in vitro mixed biofilms as well as in an in vitro cystic fibrosis lung sputum medium. Consistently, we demonstrated that production of HCN by P. aeruginosa controls S. aureus growth in a mouse model of airways coinfected by P. aeruginosa and S. aureus. Our study therefore demonstrates that P. aeruginosa HCN contributes to local and distant airborne competition against S. aureus and potentially other HCN-sensitive bacteria in contexts relevant to cystic fibrosis and other polymicrobial infectious diseases. IMPORTANCE Airborne volatile compounds produced by bacteria are often only considered attractive or repulsive scents, but they also directly contribute to bacterial physiology. Here, we showed that volatile hydrogen cyanide (HCN) released by a wide range of Pseudomonas aeruginosa strains controls Staphylococcus aureus growth in low-oxygen in vitro biofilms or aggregates and in vivo lung environments. These results are of pathophysiological relevance, since lungs of cystic fibrosis patients are known to present microaerobic areas and to be commonly associated with the presence of S. aureus and P. aeruginosa in polymicrobial communities. Our study therefore provides insights into how a bacterial volatile compound can contribute to the exclusion of S. aureus and other HCN-sensitive competitors from P. aeruginosa ecological niches. It opens new perspectives for the management or monitoring of P. aeruginosa infections in lower-lung airway infections and other polymicrobial disease contexts.

RevDate: 2022-09-21

Nemchenko UM, Sitnikova KO, Belkova NL, et al (2022)

Effects of аntimicrobials on Pseudomonas aeruginosa biofilm formation.

Vavilovskii zhurnal genetiki i selektsii, 26(5):495-501.

Pseudomonas aeruginosa is one of the most problematic pathogens in medical institutions, which may be due to the ability of this microorganism to exist in a biofilm, which increases its resistance to antimicrobials, as well as its prevalence and survival ability in the external environment. This work aimed to evaluate the antimicrobial susceptibility of P. aeruginosa strains in planktonic and biofilm forms. We studied 20 strains of P. aeruginosa collected during 2018-2021 by specialists from the Laboratory of Microbiome and Microecology of the Scientific Centre for Family Health and Human Reproduction Problems. The identification of strains was carried out using test systems for differentiating gram-negative non-fermenting bacteria (NEFERMtest 24 Erba Lachema s.r.o., Czech Republic), and confirmed by mass spectrometric analysis and 16S rRNA gene sequencing. Antimicrobial activity was assessed by the degree of inhibition of cell growth in planktonic and biofilm forms (on a flat-bottomed 96-well plastic immunological plate). All clinical isolates of P. aeruginosa were biofilm formers, 47.6 % of the isolates were weak biofilm formers, and 52.4 % of the isolates were moderate biofilm formers. Planktonic cells and the forming biofilm of the tested P. aeruginosa strains were carbapenems-resistant. Biofilm formation was suppressed in more than 90 % of cases by the agents of the cephalosporin and aminoglycoside groups. Antimicrobial susceptibility of P. aeruginosa strains in the formed biofilm was significantly lower (p < 0.05). Carbapenems and cephalosporins did not affect the mature biofilms of the tested P. aeruginosa strains in more than 60 % of cases. Only non-beta-lactam antibiotics (ciprofloxacin and amikacin) suppressed the growth of planktonic cells and destroyed the mature biofilm. The revealed differences in the effect of the tested antimicrobials on the P. aeruginosa strains biofilms correlate with resistance to a number of antibiotics. To prevent biofilm formation in the hospital strains of P. aeruginosa, the use of ceftazidime may be recommended, and antimicrobials such as ciprofloxacin and amikacin may be used to affect mature biofilms of P. aeruginosa.

RevDate: 2022-09-21

Dey A, Yadav M, Kumar D, et al (2022)

A combination therapy strategy for treating antibiotic resistant biofilm infection using a guanidinium derivative and nanoparticulate Ag(0) derived hybrid gel conjugate.

Chemical science, 13(34):10103-10118 pii:d2sc02980d.

Bacteria organized in biofilms show significant tolerance to conventional antibiotics compared to their planktonic counterparts and form the basis for chronic infections. Biofilms are composites of different types of extracellular polymeric substances that help in resisting several host-defense measures, including phagocytosis. These are increasingly being recognized as a passive virulence factor that enables many infectious diseases to proliferate and an essential contributing facet to anti-microbial resistance. Thus, inhibition and dispersion of biofilms are linked to addressing the issues associated with therapeutic challenges imposed by biofilms. This report is to address this complex issue using a self-assembled guanidinium-Ag(0) nanoparticle (AD-L@Ag(0)) hybrid gel composite for executing a combination therapy strategy for six difficult to treat biofilm-forming and multidrug-resistant bacteria. Improved efficacy was achieved primarily through effective biofilm inhibition and dispersion by the cationic guanidinium ion derivative, while Ag(0) contributes to the subsequent bactericidal activity on planktonic bacteria. Minimum Inhibitory Concentration (MIC) of the AD-L@Ag(0) formulation was tested against Acinetobacter baumannii (25 μg mL-1), Pseudomonas aeruginosa (0.78 μg mL-1), Staphylococcus aureus (0.19 μg mL-1), Klebsiella pneumoniae (0.78 μg mL-1), Escherichia coli (clinical isolate (6.25 μg mL-1)), Klebsiella pneumoniae (clinical isolate (50 μg mL-1)), Shigella flexneri (clinical isolate (0.39 μg mL-1)) and Streptococcus pneumoniae (6.25 μg mL-1). Minimum bactericidal concentration, and MBIC50 and MBIC90 (Minimum Biofilm Inhibitory Concentration at 50% and 90% reduction, respectively) were evaluated for these pathogens. All these results confirmed the efficacy of the formulation AD-L@Ag(0). Minimum Biofilm Eradication Concentration (MBEC) for the respective pathogens was examined by following the exopolysaccharide quantification method to establish its potency in inhibition of biofilm formation, as well as eradication of mature biofilms. These effects were attributed to the bactericidal effect of AD-L@Ag(0) on biofilm mass-associated bacteria. The observed efficacy of this non-cytotoxic therapeutic combination (AD-L@Ag(0)) was found to be better than that reported in the existing literature for treating extremely drug-resistant bacterial strains, as well as for reducing the bacterial infection load at a surgical site in a small animal BALB/c model. Thus, AD-L@Ag(0) could be a promising candidate for anti-microbial coatings on surgical instruments, wound dressing, tissue engineering, and medical implants.

RevDate: 2022-09-20

Aherne O, Ortiz R, Fazli MM, et al (2022)

Effects of stabilized hypochlorous acid on oral biofilm bacteria.

BMC oral health, 22(1):415.

BACKGROUND: Caries and periodontitis are amongst the most prevalent diseases worldwide, leading to pain and loss of oral function for those affected. Prevention relies heavily on mechanical removal of dental plaque biofilms but for populations where this is not achievable, alternative plaque control methods are required. With concerns over undesirable side-effects and potential bacterial resistance due to the use of chlorhexidine gluconate (CHX), new antimicrobial substances for oral use are greatly needed. Here we have investigated the antimicrobial effect of hypochlorous acid (HOCl), stabilized with acetic acid (HAc), on oral biofilms and compared it to that of CHX. Possible adverse effects of stabilized HOCl on hydroxyapatite surfaces were also examined.

METHODS: Single- and mixed-species biofilms of six common oral bacteria (Streptococcus mutans, Streptococcus gordonii, Actinomyces odontolyticus, Veillonella parvula, Parvimonas micra and Porphyromonas gingivalis) within a flow-cell model were exposed to HOCl stabilized with 0.14% or 2% HAc, pH 4.6, as well as HOCl or HAc alone. Biofilm viability was assessed in situ using confocal laser scanning microscopy following LIVE/DEAD® BacLight™ staining. In-situ quartz crystal microbalance with dissipation (QCM-D) was used to study erosion of hydroxyapatite (HA) surfaces by stabilized HOCl.

RESULTS: Low concentrations of HOCl (5 ppm), stabilized with 0.14% or 2% HAc, significantly reduced viability in multi-species biofilms representing supra- and sub-gingival oral communities, after 5 min, without causing erosion of HA surfaces. No equivalent antimicrobial effect was seen for CHX. Gram-positive and Gram-negative bacteria showed no significant differential suceptibility to stabilized HOCl.

CONCLUSIONS: At low concentrations and with exposure times which could be achieved through oral rinsing, HOCl stabilized with HAc had a robust antimicrobial activity on oral biofilms, without causing erosion of HA surfaces or affecting viability of oral keratinocytes. This substance thus appears to offer potential for prevention and/or treatment of oral biofilm-mediated diseases.

RevDate: 2022-09-20

Xia M, Zhuo N, Ren S, et al (2022)

Enterococcus faecalis rnc gene modulates its susceptibility to disinfection agents: a novel approach against biofilm.

BMC oral health, 22(1):416.

BACKGROUND: Enterococcus faecalis (E. faecalis) plays an important role in the failure of root canal treatment and refractory periapical periodontitis. As an important virulence factor of E. faecalis, extracellular polysaccharide (EPS) serves as a matrix to wrap bacteria and form biofilms. The homologous rnc gene, encoding Ribonuclease III, has been reported as a regulator of EPS synthesis. In order to develop novel anti-biofilm targets, we investigated the effects of the rnc gene on the biological characteristics of E. faecalis, and compared the biofilm tolerance towards the typical root canal irrigation agents and traditional Chinese medicine fluid Pudilan.

METHODS: E. faecalis rnc gene overexpression (rnc+) and low-expression (rnc-) strains were constructed. The growth curves of E. faecalis ATCC29212, rnc+, and rnc- strains were obtained to study the regulatory effect of the rnc gene on E. faecalis. Scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and crystal violet staining assays were performed to evaluate the morphology and composition of E. faecalis biofilms. Furthermore, the wild-type and mutant biofilms were treated with 5% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), and Pudilan. The residual viabilities of E. faecalis biofilms were evaluated using crystal violet staining and colony counting assays.

RESULTS: The results demonstrated that the rnc gene could promote bacterial growth and EPS synthesis, causing the EPS-barren biofilm morphology and low EPS/bacteria ratio. Both the rnc+ and rnc- biofilms showed increased susceptibility to the root canal irrigation agents. The 5% NaOCl group showed the highest biofilm removing effect followed by Pudilan and 2% CHX. The colony counting results showed almost complete removal of bacteria in the 5% NaOCl, 2% CHX, and Chinese medicine agents' groups.

CONCLUSIONS: This study concluded that the rnc gene could positively regulate bacterial proliferation, EPS synthesis, and biofilm formation in E. faecalis. The rnc mutation caused an increase in the disinfectant sensitivity of biofilm, indicating a potential anti-biofilm target. In addition, Pudilan exhibited an excellent ability to remove E. faecalis biofilm.

RevDate: 2022-09-20

Flemming HC, van Hullebusch ED, Neu TR, et al (2022)

The biofilm matrix: multitasking in a shared space.

Nature reviews. Microbiology [Epub ahead of print].

The biofilm matrix can be considered to be a shared space for the encased microbial cells, comprising a wide variety of extracellular polymeric substances (EPS), such as polysaccharides, proteins, amyloids, lipids and extracellular DNA (eDNA), as well as membrane vesicles and humic-like microbially derived refractory substances. EPS are dynamic in space and time and their components interact in complex ways, fulfilling various functions: to stabilize the matrix, acquire nutrients, retain and protect eDNA or exoenzymes, or offer sorption sites for ions and hydrophobic substances. The retention of exoenzymes effectively renders the biofilm matrix an external digestion system influencing the global turnover of biopolymers, considering the ubiquitous relevance of biofilms. Physico-chemical and biological interactions and environmental conditions enable biofilm systems to morph into films, microcolonies and macrocolonies, films, ridges, ripples, columns, pellicles, bubbles, mushrooms and suspended aggregates - in response to the very diverse conditions confronting a particular biofilm community. Assembly and dynamics of the matrix are mostly coordinated by secondary messengers, signalling molecules or small RNAs, in both medically relevant and environmental biofilms. Fully deciphering how bacteria provide structure to the matrix, and thus facilitate and benefit from extracellular reactions, remains the challenge for future biofilm research.

RevDate: 2022-09-20

Figueroa-Valenzuela C, Montes-García JF, Vazquez-Cruz C, et al (2022)

Mannheimia haemolytica OmpH binds fibrinogen and fibronectin and participates in biofilm formation.

Microbial pathogenesis pii:S0882-4010(22)00401-6 [Epub ahead of print].

Mannheimia haemolytica is the causal agent of the shipping fever in bovines and produces high economic losses worldwide. This bacterium possesses different virulence attributes to achieve a successful infection. One of the main virulence factors expressed by a pathogen is through adhesion molecules; however, the components participating in this process are not totally known. The present work identified a M. haemolytica 41 kDa outer membrane protein (Omp) that participates in bacterial adhesion. This protein showed 100% identity with the OmpH from M. haemolytica as determined by mass spectrometry and it interacts with sheep fibrinogen. The 41 kDa M. haemolytica OmpH interacts with bovine monocytes; a previous incubation of M. haemolytica with a rabbit hyperimmune serum against this Omp diminished 45% cell adhesion. The OmpH was recognized by serum from bovines affected by acute or chronic pneumonia, indicating its in vivo expression; moreover, it showed immune cross-reaction with the serum of rabbit infected with Pasteurella multocida. The OmpH is present in biofilms and previous incubation of M. haemolytca with rabbit serum against this protein diminished biofilm, indicating this protein's participation in biofilm formation. M. haemolytica OmpH is proposed as a relevant immunogen in bovine pneumonia protection.

RevDate: 2022-09-20

Farooq AJ, Chamberlain M, Poonja A, et al (2022)

Peaks, pores, and dragon eggs: Uncovering and quantifying the heterogeneity of treatment wetland biofilm matrices.

The Science of the total environment pii:S0048-9697(22)05956-3 [Epub ahead of print].

Biofilms serve to house diverse microbial communities, which are responsible for the majority of wastewater constituent degradation and transformation in treatment wetlands (TWs). TW biofilm has been generally conceptualized as a relatively uniform film covering available surfaces. However, no direct visual 3D representation of biofilm morphology in TWs has been conducted. This study focuses on imaging the morphology of detached, gravel-associated, and rhizospheric (Phalaris arundinacea) biofilms from subsurface TW mesocosms. Images obtained through both traditional light microscopy, environmental scanning electron microscopy (E-SEM) and Wet-SEM revealed that TW biofilms are structurally heterogeneous ranging from corrugated films to clusters of aggregates. Features such as water channels and pores were observed suggesting that pollutant transport inside biofilms is complex, and that the interfacial surface area between water and biofilm is much larger than previously understood. Biofilm thickness generally ranged between 170 and 240 μm, with internal biofilm porosities estimated as 34 ± 10 %, reaching a maximum of 50 %. Internal biofilm matrix pore diameters ranged from 1 to 205.2 μm, with a distribution that favored pores and channels smaller than 10 μm, and a mean equivalent spherical diameter of 8.6 μm. Based on the large variation in pore and channel sizes it is expected that a variety of flow regimes and therefore pollutant dynamics are likely to occur inside TW biofilm matrices. Based on the visual evidence and analysis, a new conceptual model was created to reflect the microscale TW biofilm dynamics and morphology. This new conceptual model will serve to inform future biokinetic modelling, microscale hydrology, microbial community assessment, and pollutant treatment studies.

RevDate: 2022-09-20

Chakroun I, Fedhila K, Maatallah M, et al (2022)

The Synbiotic Effect of Probiotics and Dried Spirulina platensis or Phycocyanin on Biofilm Formation by Salmonella Typhimurium and Staphylococcus aureus.

Foodborne pathogens and disease [Epub ahead of print].

This study aimed to evaluate the synbiotic effect of probiotics and dried Spirulina platensis or phycocyanin on autoaggregation, coaggregation, and the inhibition of biofilm formation by Salmonella Typhimurium and Staphylococcus aureus on 96-well microtiter plates and Human colon carcinoma cells-116 surfaces. The results showed that the probiotics strains cultured in the presence of S. platensis exhibited the highest autoaggregation values, ranging between 68.5 and 74.2% after 24 h. All probiotic strains with or without S. platensis and phycocyanin showed coaggregation abilities with S. Typhimurium and S. aureus. Interestingly, significant effect of S. platensis and phycocyanin supplementation was observed on the inhibition of the biofilm formation by the selected pathogens during the competition, exclusion, and displacement on abiotic and biotic surfaces.

RevDate: 2022-09-21

Dos Santos RL, Davanzo EFA, Palma JM, et al (2022)

Molecular characterization and biofilm-formation analysis of Listeria monocytogenes, Salmonella spp., and Escherichia coli isolated from Brazilian swine slaughterhouses.

PloS one, 17(9):e0274636 pii:PONE-D-22-10849.

This study aimed to verify the presence of Listeria monocytogenes, Salmonella spp., and Escherichia coli in two Brazilian swine slaughterhouses, as well as to perform antibiograms, detect virulence and antimicrobial resistance genes, and evaluate the in vitro biofilm-forming capability of bacterial isolates from these environments. One Salmonella Typhi isolate and 21 E. coli isolates were detected, while L. monocytogenes was not detected. S. Typhi was isolated from the carcass cooling chamber's floor, resistant to several antimicrobials, including nalidixic acid, cefazolin, chloramphenicol, doxycycline, streptomycin, gentamicin, tetracycline, and sulfonamide, and contained resistance genes, such as tet(B), tet(C), tet(M), and ampC. It also showed moderate biofilm-forming capacity at 37°C after incubating for 72 h. The prevalence of the 21 E. coli isolates was also the highest on the carcass cooling chamber floor (three of the four samplings [75%]). The E. coli isolates were resistant to 12 of the 13 tested antimicrobials, and none showed sensitivity to chloramphenicol, an antimicrobial prohibited in animal feed since 2003 in Brazil. The resistance genes MCR-1, MCR-3, sul1, ampC, clmA, cat1, tet(A), tet(B), and blaSHV, as well as the virulence genes stx-1, hlyA, eae, tir α, tir β, tir γ, and saa were detected in the E. coli isolates. Moreover, 5 (23.8%) and 15 (71.4%) E. coli isolates presented strong and moderate biofilm-forming capacity, respectively. In general, the biofilm-forming capacity increased after incubating for 72 h at 10°C. The biofilm-forming capacity was the lowest after incubating for 24 h at 37°C. Due to the presence of resistance and virulence genes, multi-antimicrobial resistance, and biofilm-forming capacity, the results of this study suggest a risk to the public health as these pathogens are associated with foodborne diseases, which emphasizes the hazard of resistance gene propagation in the environment.

RevDate: 2022-09-20

Keikha M, M Karbalaei (2022)

Biofilm formation status in ESBL-producing bacteria recovered from clinical specimens of patients: A systematic review and meta-analysis.

BACKGROUND: Recently, the emergence and spread of extended-spectrum beta-lactamase (ESBL) bacteria have become a global health concern. In addition, the ability to form biofilm due to less impermeability to antibiotics and the horizontal transformation (conjugation) of genes involved in antibiotic resistance have exacerbated the concerns. With a comprehensive meta-analysis, this study evaluated the potential relationship between ESBL and biofilm formation.

METHODS: A literature search was performed using global databases, such as PubMed and Scopus, up to November 2021. We retrieved all relevant documents and selected eligible articles based on inclusion criteria. Finally, the potential association between the biofilm formation capacity and resistance of ESBL-producing bacteria was measured with an odds ratio and a 95% confidence interval.

RESULTS: In the present study, 17 articles, including 2,069 Gram-negative isolates, were considered as eligible. The prevalence of biofilm formation in all clinical isolates of ESBL and non-ESBL pathogens was 72.4% (95% CI: 60.7-81.6) and 40.5% (95% CI: 30.2-51.8), respectively. Our results showed a positive relationship between the ability for biofilm formation and conferring antibiotic resistance in ESBL-producing bacteria (OR: 3.35; 95% CI: 1.67-6.74; p-value: 0.001).

CONCLUSION: In general, we showed the rate of biofilm formation to be significantly higher in ESBLproducing strains. Given the current results, the updated therapeutic guidelines should consider the role of biofilm production for optimal therapy, treatment course, and clinical outcomes rather than the recommendation of antimicrobial agents by focusing on the results of the antibiotic susceptibility test.

RevDate: 2022-09-21

Nercessian D, JP Busalmen (2022)

Cell Adhesion and Biofilm Formation Analysis.

Methods in molecular biology (Clifton, N.J.), 2522:407-417.

Cell adhesion to surfaces and ulterior biofilm formation are critical processes in microbial development since living in biofilms is the preferred way of life within microorganisms. These processes are known to influence not only microorganisms development in the environment, but also their participation in biotechnological processes and have been the focus of intense research that as a matter of fact, was mainly directed to the bacterial domain. Archaea also adhere to surfaces and have been shown forming biofilms, but studies performed until present did not exploit the diversity of methods probed to be useful along bacterial biofilm research.An experimental setup is described here with the aim of stimulating archaeal biofilm research. It can be used for studying cell adhesion and biofilm formation under controlled flow conditions and allows performing in situ optical microscopy (phase contrast, fluorescence, or confocal) and/or spectroscopic techniques (UV-Vis, IR, or Raman) to determine structural and functional biofilm features and their evolution in time. Variants are described with specific aims as working in anaerobiosis and allow sampling of biological material along time.

RevDate: 2022-09-21

Mutan Z, Schiller H, Schulze S, et al (2022)

Immersed Liquid Biofilm and Honeycomb Pattern Formations in Haloferax volcanii.

Methods in molecular biology (Clifton, N.J.), 2522:387-395.

Biofilms are cellular aggregates encased in extracellular polymeric substances and are commonly formed by single-celled eukaryotes, bacteria, and archaea. In addition to attaching to solid surfaces, these cellular aggregates can also be observed floating on or immersed within liquid cultures. While biofilms on surfaces have been studied in some archaea, little is known about liquid biofilms. Surprisingly, immersed liquid biofilms of the model archaeon Haloferax volcanii do not require the same set of machinery needed to form surface-attached biofilms. In fact, to date not a single gene has been identified that is involved in forming immersed liquid biofilms. Interestingly, after an immersed liquid biofilm forms, removal of the Petri dish lid induces rapid, transient, and reproducible honeycomb patterns within the immersed liquid biofilm itself, triggered by a reduction in humidity. In this chapter, we outline a protocol for both immersed liquid biofilm and honeycomb pattern formations. This protocol will be essential for determining the novel components required for the formation of immersed liquid biofilms and honeycomb patterns.

RevDate: 2022-09-21

Oluwole OM (2022)


African journal of infectious diseases, 16(2 Suppl):59-71.

Biofilm formation, especially on indwelling medical devices such as catheters, can result in infections and substantially affect patients' quality of life. Biofilm-associated infections have led to increased morbidity and mortality, increased cost of treatment, and length of hospital stay. However, all of the identified consequences of the biofilm-associated infections had been attributed to the reduced susceptibility of biofilm to conventional antimicrobial agents which has necessitated the development of a new strategy for biofilm infections control, thereby making a search for more effective antimicrobial agents from plant source inevitable. So far, some antimicrobial agents (crude or isolated compounds) from plant sources affect a specific stage of biofilm development while a few of them have been developed into a suitable dosage form for biofilm control. In this review, an attempt is made to look into some definitions of biofilm by "biofilmologists", stages in biofilm formation, mechanisms of resistance in biofilm, biofilm control strategies, the use of some natural products in biofilm control and concepts of probiotics as agents of biofilm control.

RevDate: 2022-09-19

Purtschert-Montenegro G, Cárcamo-Oyarce G, Pinto-Carbó M, et al (2022)

Pseudomonas putida mediates bacterial killing, biofilm invasion and biocontrol with a type IVB secretion system.

Nature microbiology [Epub ahead of print].

Many bacteria utilize contact-dependent killing machineries to eliminate rivals in their environmental niches. Here we show that the plant root colonizer Pseudomonas putida strain IsoF is able to kill a wide range of soil and plant-associated Gram-negative bacteria with the aid of a type IVB secretion system (T4BSS) that delivers a toxic effector into bacterial competitors in a contact-dependent manner. This extends the range of targets of T4BSSs-so far thought to transfer effectors only into eukaryotic cells-to prokaryotes. Bioinformatic and genetic analyses showed that this killing machine is entirely encoded by the kib gene cluster located within a rare genomic island, which was recently acquired by horizontal gene transfer. P. putida IsoF utilizes this secretion system not only as a defensive weapon to kill bacterial competitors but also as an offensive weapon to invade existing biofilms, allowing the strain to persist in its natural environment. Furthermore, we show that strain IsoF can protect tomato plants against the phytopathogen Ralstonia solanacearum in a T4BSS-dependent manner, suggesting that IsoF can be exploited for pest control and sustainable agriculture.

RevDate: 2022-09-19

Allkja J, Goeres DM, Azevedo AS, et al (2022)

"Interactions of microorganisms within a urinary catheter polymicrobial biofilm model".

Biotechnology and bioengineering [Epub ahead of print].

Biofilms are often polymicrobial in nature, which can impact their behaviour and overall structure, often resulting in an increase in biomass and enhanced antimicrobial resistance. Using plate counts and locked nucleic acid/2'-O-methyl-RNA fluorescence in situ hybridization (LNA/2'OMe-FISH), we studied the interactions of four species commonly associated with catheter associated urinary tract infections (CAUTI): Enterococcus faecalis, Escherichia coli, Candida albicans and Proteus mirabilis. Eleven combinations of biofilms were grown on silicone coupons placed in 24-well plates for 24 hrs, 37 ºC, in artificial urine medium (AUM). Results showed that P. mirabilis was the dominant species and was able to inhibit both E. coli and C. albicans growth. In the absence of P. mirabilis, an antagonistic relationship between E. coli and C. albicans was observed, with the former being dominant. E. faecalis growth was not affected in any combination, showing a more mutualistic relationship with the other species. Imaging results correlated with the plate count data and provided visual verification of species undetected using the viable plate count. Moreover, the three bacterial species showed overall good repeatability SD (Sr) values (0.1 - 0.54) in all combinations tested, whereas C. albicans had higher repeatability Sr values (0.36-1.18). The study showed the complexity of early-stage interactions in polymicrobial biofilms. These interactions could serve as a starting point when considering targets for preventing or treating CAUTI biofilms containing these species. This article is protected by copyright. All rights reserved.

RevDate: 2022-09-21

Rathore SS, Cheepurupalli L, Gangwar J, et al (2022)

Biofilm of Klebsiella pneumoniae minimize phagocytosis and cytokine expression by macrophage cell line.

AMB Express, 12(1):122.

Infectious bacteria in biofilm mode are involved in many persistent infections. Owing to its importance in clinical settings, many in vitro and in vivo studies are being conducted to study the structural and functional properties of biofilms, their drug resistant mechanism and the s urvival mechanism of planktonic and biofilm cells. In this regard, there is not sufficient information on the interaction between Klebsiella biofilm and macrophages. In this study, we have attempted to unravel the interaction between Klebsiella biofilm and macrophages in terms of phagocytic response and cytokine expression. In vitro phagocytosis assays were performed for heat inactivated and live biofilms of K. pneumoniae, together with the expression analysis of TLR2, iNOS, inflammatory cytokines such as IL-β1, IFN-γ, IL-6, IL-12, IL-4, TNF-α and anti-inflammatory cytokine, IL-10. A phagocytic rate of an average of 15% was observed against both heat inactivated and live biofilms when LPS + IFN-γ activated macrophages were used. This was significantly higher than non-activated macrophages when tested against heat inactivated and live biofilms (average 8%). Heat-inactivated and live biofilms induced similar phagocytic responses and up-regulation of pro-inflammatory genes in macrophages, indirectly conveying that macrophage responses are to some extent dependent on the biofilm matrix.

RevDate: 2022-09-20

González-Paz JR, Becerril-Varela K, C Guerrero-Barajas (2022)

Iron reducing sludge as a source of electroactive bacteria: assessing iron reduction in biofilm bacteria, planktonic cells and isolates from a microbial fuel cell.

Archives of microbiology, 204(10):632.

In this study, bacteria from a microbial fuel cell (MFC) and isolates were evaluated on their Fe3+ reduction capability at different concentrations of iron using acetate as the sole source of carbon. The results demonstrated that the planktonic cells can reach an iron reduction up to 60% at 27 mmol Fe3+. Azospira oryzae (µ 0.89 ± 0.27 d-1) and Cupriavidus metallidurans CH34 (µ 2.34 ± 0.81 d-1) presented 55 and 62% of Fe3+ reduction, respectively, at 16 mmol l-1. Enterobacter bugandensis (µ 0.4 ± 0.01 d-1) 40% Fe3+ at 27 mmol l-1, Citrobacter freundii ATCC 8090 (µ 0.23 ± 0.05 d-1) and Citrobacter murliniae CDC2970-59 (µ 0.34 ± 0.02 d-1) reduced Fe3+ in ~ 50%, at 55 mmol l-1. This is the first report on these bacteria on a percentage of iron reduction. These results may be useful for anode design to contribute to a higher energy generation in MFCs.

RevDate: 2022-09-19

Gill SP, Hunter WR, Coulson LE, et al (2022)

Synthetic and biological surfactant effects on freshwater biofilm community composition and metabolic activity.

Applied microbiology and biotechnology [Epub ahead of print].

Surfactants are used to control microbial biofilms in industrial and medical settings. Their known toxicity on aquatic biota, and their longevity in the environment, has encouraged research on biodegradable alternatives such as rhamnolipids. While previous research has investigated the effects of biological surfactants on single species biofilms, there remains a lack of information regarding the effects of synthetic and biological surfactants in freshwater ecosystems. We conducted a mesocosm experiment to test how the surfactant sodium dodecyl sulfate (SDS) and the biological surfactant rhamnolipid altered community composition and metabolic activity of freshwater biofilms. Biofilms were cultured in the flumes using lake water from Lake Lunz in Austria, under high (300 ppm) and low (150 ppm) concentrations of either surfactant over a four-week period. Our results show that both surfactants significantly affected microbial diversity. Up to 36% of microbial operational taxonomic units were lost after surfactant exposure. Rhamnolipid exposure also increased the production of the extracellular enzymes, leucine aminopeptidase, and glucosidase, while SDS exposure reduced leucine aminopeptidase and glucosidase. This study demonstrates that exposure of freshwater biofilms to chemical and biological surfactants caused a reduction of microbial diversity and changes in biofilm metabolism, exemplified by shifts in extracellular enzyme activities. KEY POINTS: • Microbial biofilm diversity decreased significantly after surfactant exposure. • Exposure to either surfactant altered extracellular enzyme activity. • Overall metabolic activity was not altered, suggesting functional redundancy.

RevDate: 2022-09-19

Tomlinson BR, Denham GA, Torres NJ, et al (2022)

Assessing the Role of Cold-Shock Protein C: a Novel Regulator of Acinetobacter baumannii Biofilm Formation and Virulence.

Infection and immunity [Epub ahead of print].

Acinetobacter baumannii is a formidable opportunistic pathogen that is notoriously difficult to eradicate from hospital settings. This resilience is often attributed to a proclivity for biofilm formation, which facilitates a higher tolerance toward external stress, desiccation, and antimicrobials. Despite this, little is known regarding the mechanisms orchestrating A. baumannii biofilm formation. Here, we performed RNA sequencing (RNA-seq) on biofilm and planktonic populations for the multidrug-resistant isolate AB5075 and identified 438 genes with altered expression. To assess the potential role of genes upregulated within biofilms, we tested the biofilm-forming capacity of their respective mutants from an A. baumannii transposon library. In so doing, we uncovered 24 genes whose disruption led to reduced biofilm formation. One such element, cold shock protein C (cspC), had a highly mucoid colony phenotype, enhanced tolerance to polysaccharide degradation, altered antibiotic tolerance, and diminished adherence to abiotic surfaces. RNA-seq of the cspC mutant revealed 201 genes with altered expression, including the downregulation of pili and fimbria genes and the upregulation of multidrug efflux pumps. Using transcriptional arrest assays, it appears that CspC mediates its effects, at least in part, through RNA chaperone activity, influencing the half-life of several important transcripts. Finally, we show that CspC is required for survival during challenge by the human immune system and is key for A. baumannii dissemination and/or colonization during systemic infection. Collectively, our work identifies a cadre of new biofilm-associated genes within A. baumannii and provides unique insight into the global regulatory network of this emerging human pathogen.

RevDate: 2022-09-20

Ghidini S, De Luca S, Rodríguez-López P, et al (2022)

Microbial contamination, antimicrobial resistance and biofilm formation of bacteria isolated from a high-throughput pig abattoir.

Italian journal of food safety, 11(3):10160.

The aim of this work was to assess the level of microbial contamination and resistance of bacteria isolated from a highthroughput heavy pig slaughterhouse (approx. 4600 pigs/day) towards antimicrobials considered as critical for human, veterinary or both chemotherapies. Samples, pre-operative and operative, were obtained in 4 different surveys. These comprised environmental sampling, i.e. air (ntotal = 192) and surfaces (ntotal = 32), in four different locations. Moreover, a total of 40 carcasses were sampled in two different moments of slaughtering following Reg. (CE) 2073/2005. Overall, 60 different colonies were randomly selected from VRBGA plates belonging to 20 species, 15 genera and 10 families being Enterobacteriaceae, Moraxellaceae and Pseudomonadaceae the most represented ones. Thirty-seven isolates presented resistance to at least one molecule and seventeen were classified as multi-drug resistant. Enterobacteriaceae, particularly E. coli, displayed high MIC values towards trimethoprim, ampicillin, tetracycline and sulphametoxazole with MICmax of 16, 32, 32 and 512 mg/L, respectively. Moreover, isolated Pseudomonas spp. showed high MIC values in critical antibiotics such as ampicillin and azithromycin with MICmax of 32 and 64 mg/L, respectively. Additionally, in vitro biofilm formation assays demonstrated that fifteen of these isolates can be classified as strong biofilm formers. Results demonstrated that a high diversity of bacteria containing antibiotic resistant and multiresistant species is present in the sampled abattoir. Considering these findings, it could be hypothesised that the processing environment could be a potential diffusion determinant of antibiotic resistant bacteria through the food chain and operators.

RevDate: 2022-09-20

Jia F, Sun MY, Zhang XJ, et al (2020)

Total alkaloids of Sophora alopecuroides- and matrine-induced reactive oxygen species impair biofilm formation of Staphylococcus epidermidis and increase bacterial susceptibility to ciprofloxacin.

Chinese herbal medicines, 12(4):390-398.

Objective: To investigate the mechanism by which total alkaloids of Sophora alopecuroides (TASA) and matrine (MT) impair biofilm to increase the susceptibility of Staphylococcus epidermidis (S. epidermidis) to ciprofloxacin.

Methods: The minimum biofilm inhibitory concentration (mBIC) was determined using a 2-fold dilution method. Structure of biofilm of S. epidermidis was examined by Confocal Laser Scanning Microscope (CLSM). The cellular reactive oxygen species (ROS) was determined using a DCFH-DA assay. The key factors related to the regulation of ROS were accessed using respective kits.

Results: TASA and MT were more beneficial to impair biofilm of S. epidermidis than ciprofloxacin (CIP) (P < 0.05). TASA and MT were not easily developed resistance to biofilm-producing S. epidermidis. The mBIC of CIP decreased by 2-6-fold following the treatment of sub-biofilm inhibitory concentration (sub-BIC) TASA and MT, whereas the mBIC of CIP increased by 2-fold following a treatment of sub-BIC CIP from the first to sixth generations. TASA and MT can improve the production of ROS in biofilm-producing S. epidermidis. The ROS content was decreased 23%-33% following the treatment of sub-mBIC CIP, whereas ROS content increased 7%-24% following treatment with TASA + CIP and MT + CIP combination from the first to sixth generations. Nitric oxide (NO) as a ROS, which was consistent with the previously confirmed relationship between ROS and drug resistance. Related regulatory factors-superoxide dismutase (SOD) and glutathione peroxidase (GSH) could synergistically maintain the redox balance in vivo.

Conclusion: TASA and MT enhanced reactive oxygen species to restore the susceptibility of S. epidermidis to ciprofloxacin.

RevDate: 2022-09-21
CmpDate: 2022-09-20

Amer MA, Ramadan MA, Attia AS, et al (2022)

Silicone Foley catheters impregnated with microbial indole derivatives inhibit crystalline biofilm formation by Proteus mirabilis.

Frontiers in cellular and infection microbiology, 12:1010625.

Proteus mirabilis is a common causative agent for catheter-associated urinary tract infections (CAUTI). The crystalline biofilm formation by P. mirabilis causes catheter encrustation and blockage leading to antibiotic treatment resistance. Thus, biofilm formation inhibition on catheters becomes a promising alternative for conventional antimicrobial-based treatment that is associated with rapid resistance development. Our previous work has demonstrated the in vitro antibiofilm activity of microbial indole derivatives against clinical isolates of P. mirabilis. Accordingly, we aim to evaluate the capacity of silicone Foley catheters (SFC) impregnated with these indole derivatives to resist biofilm formation by P. mirabilis both phenotypically and on the gene expression level. Silicon Foley catheter was impregnated with indole extract recovered from the supernatant of the rhizobacterium Enterobacter sp. Zch127 and the antibiofilm activity was determined against P. mirabilis (ATCC 12435) and clinical isolate P8 cultured in artificial urine. The indole extract at sub-minimum inhibitory concentration (sub-MIC=0.5X MIC) caused a reduction in biofilm formation as exhibited by a 60-70% reduction in biomass and three log10 in adhered bacteria. Results were confirmed by visualization by scanning electron microscope. Moreover, changes in the relative gene expression of the virulence genes confirmed the antibiofilm activity of the indole extract against P. mirabilis. Differential gene expression analysis showed that extract Zch127 at its sub-MIC concentration significantly down-regulated genes associated with swarming activity: umoC, flhC, flhD, flhDC, and mrpA (p< 0.001). In addition, Zch127 extract significantly down-regulated genes associated with polyamine synthesis: speB and glnA (p< 0.001), as well as the luxS gene associated with quorum sensing. Regulatory genes for capsular polysaccharide formation; rcsB and rcsD were not significantly affected by the presence of the indole derivatives. Furthermore, the impregnated catheters and the indole extract showed minimal or no cytotoxic effect against human fibroblast cell lines indicating the safety of this intervention. Thus, the indole-impregnated catheter is proposed to act as a suitable and safe strategy for reducing P. mirabilis CAUTIs.

RevDate: 2022-09-20

Huang K, Lin B, Liu Y, et al (2022)

Correlation Analysis between Chronic Osteomyelitis and Bacterial Biofilm.

Stem cells international, 2022:9433847.

Objective: To study the role of bacterial biofilm (BBF) in the formation of chronic osteomyelitis and its prevention and treatment.

Methods: In this paper, a large amount of relevant literature was searched for analysis and summary, and the key words "chronic osteomyelitis," "bacterial biofilm," "infection," and "debridement" were searched in databases, mainly CNKI, Wanfang, and Wipu. The search was conducted until December 2020. The role of bacterial biofilm formation in chronic osteomyelitis and its prevention were analyzed.

Results: Chronic osteomyelitis is formed mainly due to poor blood supply and drug-resistant bacteria, of which cellular biofilm is the most important cause. BBF forms on the surface of necrotic soft tissue and bone tissue, which has a protective effect on bacteria and greatly enhances their resistance to antibiotics, leading to difficulties in complete bacterial clearance and recurrent infections in osteomyelitis.

Conclusion: Through an in-depth study of the molecular biology and signal transduction of osteomyelitis biofilm, antibiotic biofilm treatment strategies and surgical debridement remain the focus of clinical translation of chronic osteomyelitis.

RevDate: 2022-09-20

Eckert JA, Rosenberg M, Rhen M, et al (2022)

An optotracer-based antibiotic susceptibility test specifically targeting the biofilm lifestyle of Salmonella.

Biofilm, 4:100083.

Antimicrobial resistance is a medical threat of global dimensions. Proper antimicrobial susceptibility testing (AST) for drug development, patient diagnosis and treatment is crucial to counteract ineffective drug use and resistance development. Despite the important role of bacterial biofilms in chronic and device-associated infections, the efficacy of antibiotics is determined using planktonic cultures. To address the need for antibiotics targeting bacteria in the biofilm lifestyle, we here present an optotracing-based biofilm-AST using Salmonella as model. Our non-disruptive method enables real-time recording of the extracellular matrix (ECM) components, providing specific detection of the biofilm lifestyle. Biofilm formation prior to antibiotic challenge can thus be confirmed and pre-treatment data collected. By introducing Kirby-Bauer discs, we performed a broad screen of the effects of antibiotics representing multiple classes, and identified compounds with ECM inhibitory as well as promoting effects. These compounds were further tested in agar-based dose-response biofilm-AST assays. By quantifying the ECM based on the amount of curli, and by visualizing the biofilm size and morphology, we achieved new information directly reflecting the treated biofilm. This verified the efficacy of several antibiotics that were effective in eradicating pre-formed biofilms, and it uncovered intriguing possible resistance mechanisms initiated in response to treatments. By providing deeper insights into the resistances and susceptibilities of microbes, expanded use of the biofilm-AST will contribute to more effective treatments of infections and reduced resistance development.

RevDate: 2022-09-18

Zhang W, Wu Y, Wu J, et al (2022)

Enhanced removal of sulfur-containing organic pollutants from actual wastewater by biofilm reactor: Insights of sulfur transformation and bacterial metabolic traits.

Environmental pollution (Barking, Essex : 1987) pii:S0269-7491(22)01401-4 [Epub ahead of print].

Sulfur-containing organic pollutants in wastewater could threaten human health due to their high malodor and toxicity, and their conversion processes are more complex than inorganic sulfur compounds. Membrane aerated biofilm reactor (MABR), as a novel and environmentally-friendly biofilm-based technology, is able to remove inorganic sulfur in synthetic wastewater. However, it is unknown how sulfur-containing organic pollutants in actual wastewater are transformed in MABR system. This work demonstrated the feasibility of MABR to eliminate sulfur-containing organic pollutants in actual wastewater, and the removal efficiency could be reached at approximately 100%. Meanwhile, over 70% of sulfur-containing organic contaminants were transformed to SO42- during the long-term operation. Further analysis indicated that the functional bacteria that participated in sulfur transformation and carbohydrates degradation (e.g., Chujaibacter, Microscillaceae sp., and Thiobacillus) were evidently enriched when treating actual wastewater. Moreover, the critical metabolic pathways (e.g., sulfur metabolism, glycolysis metabolism, and pyruvate metabolism), and the corresponding genetic expressions (e.g., nrrA, tauA, tauC, sorA, and SUOX) were evidently up-regulated during long-term operation, which was beneficial for the transformation of sulfur-containing organic pollutants in actual wastewater by MABR. This work would expand the application of MABR for treating the actual sulfur-containing organic wastewater and provide an in-depth understanding of the organic sulfur transformation in MABR.

RevDate: 2022-09-18

Nie LJ, Ye WQ, Xie WY, et al (2022)

Biofilm: New insights in the biological control of fruits with Bacillus amyloliquefaciens B4.

Microbiological research, 265:127196 pii:S0944-5013(22)00236-1 [Epub ahead of print].

Biofilms are sessile microbial communities growing on surfaces, which are encased in some self-produced extracellular material. Beneficial biofilm could be widely used in agriculture, food, medicine, environment and other fields. As an ideal biocontrol agent, Bacillus amyloliquefaciens B4 can form a strong biofilm under static conditions. In this study, we screened out metal compounds that enhanced or inhibited the biofilm formation ability of B4, established the relationship between the biofilm of B4 strain and its postharvest biocontrol effect, and explored the regulation of metal compounds on the biofilm formation. The results showed 0.5 mmol L-1 ferric chloride could enhance the biofilm formation and strengthen the antifungal effect of B4, indicating that there was a positive relationship between the growth of biofilm and its biocontrol effect. The enhanced biofilm had a certain biocontrol effect on different fruit, including peach, loquat, Kyoho grape and cherry tomato. Furthermore, the expression of degU and tasA was affected by metal ion treatment, which meant the genes might be essential for the biofilm formation of B4. Our findings suggested that biofilm of B. amyloliquefaciens played an essential role in the process of biocontrol and it might be a novel strategy for managing postharvest fruit decay.

RevDate: 2022-09-17

Xie T, Liu X, Xu Y, et al (2022)

Coupling methanotrophic denitrification to anammox in a moving bed biofilm reactor for nitrogen removal under hypoxic conditions.

The Science of the total environment pii:S0048-9697(22)05894-6 [Epub ahead of print].

Simultaneous removal of ammonium and nitrate was achieved in a methane-fed moving bed biofilm reactor (MBBR). In the reactor, methanotrophic microorganisms oxidized methane under hypoxic conditions likely to methanol, hence providing an electron donor to denitrifiers to reduce nitrate to nitrite that then allowed anaerobic ammonium oxidizing bacteria (Anammox) to remove excess ammonium as N2. The ammonium and nitrate removal rates reached 72.09 ± 5.81 mgNH4+-N/L/d and 62.61 ± 4.17 mgNO3--N/L/d when the MBBR was operated in continuous mode. Nitrate removal by the methane-fed mixed consortia was confirmed in a batch test revealing a CH4/NO3- molar removal ratio of 1.15. The functional populations were unveiled by FISH analysis and 16S rRNA gene sequencing, which showed that the biofilm was dominated by Anammox bacteria (Candidatus Kuenenia) and diverse taxa associated with the capacity for denitrification: aerobic methanotrophs (Methylobacter, Methylomonas, and unclassified Methylococcaceae), methylotrophic denitrifiers (Opitutaceae and Methylophilaceae), and other heterotrophic denitrifiers (Ignavibacteriaceae, Anaerolineaceae, Comamonadaceae, Rhodocyclaceae and Thauera). Neither DAMO archaea nor DAMO bacteria were found in the sequencing analysis, indicating that more unknown community members possess the metabolic capacity of methanotrophic denitrification.

RevDate: 2022-09-20
CmpDate: 2022-09-20

Harikrishnan P, Arayambath B, Jayaraman VK, et al (2022)

Thidiazuron, a phenyl-urea cytokinin, inhibits ergosterol synthesis and attenuates biofilm formation of Candida albicans.

World journal of microbiology & biotechnology, 38(12):224.

Candida albicans is a common human fungal pathogen that colonizes mucosa and develops biofilm in the oral cavity that causes oral candidiasis. It has been reported that cytochrome P450 enzyme (CYP51), a vital part of the ergosterol synthesis cascade, is associated with Candida infections and its biofilm formation. Thidiazuron, a phenyl-urea cytokinin, exhibits anti-senescence and elicitor activity against fungal infection in plants. However, how Thidiazuron impacts C. albicans biofilm formation is still uncertain. Here, we aimed to investigate the effects of a Thidiazuron against the growth and biofilm formation properties of C. albicans using in silico and in vitro experimental approaches. A preliminary molecular docking study revealed potential interaction between Thidiazuron and amino acid residues of CYP51. Further in vitro antifungal susceptibility test, scanning electron microscopy (SEM) and time kill analysis revealed the anti-fungal activity of Thidiazuron in both dose and time-dependent manner. Crystal violet staining, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay revealed 50% inhibition in C. albicans biofilm by Thidiazuron at concentrations 11 and 19 µM respectively. Acridine orange staining assay visually confirmed the biofilm inhibitory potential of Thidiazuron. The gene expression study showed that Thidiazuron treatment down regulated the expression of genes involved in ergosterol synthesis (ERG3, ERG11, ERG25), cell adhesion (ASL3, EAP1), and hyphae development (EFG1, HWP1, SAP5) in C. albicans. Wherease, the expression of negative transcription regulator of hyphae (NRG1) was upregulated (5.7-fold) by Thidiazuron treatment. Collectively, our data suggest that Thidiazuron is a robust antifungal compound and an outstanding biofilm inhibitor, which may promise further therapeutic development due to CYP51 binding and inhibition of ergosterol formation against C. albicans.

RevDate: 2022-09-20
CmpDate: 2022-09-20

Boltz JP, GT Daigger (2022)

A mobile-organic biofilm process for wastewater treatment.

Water environment research : a research publication of the Water Environment Federation, 94(9):e10792.

The mobile-organic biofilm (MOB) process includes mobile biofilms and their retention screens with a bioreactor and liquid and solid separation. The MOB process is inexpensive and easy to integrate with wastewater treatment (WWT) processes, and it provides for high-rate WWT in biofilm or hybrid bioreactors. This paper describes three modes of MOB process operation. The first mode of operation, Mode I, has a mobile-biofilm reactor and a mobile-biofilm retention screen that is downstream of and external to a bioreactor and upstream of liquid and solid separation. Modes II and III have a hybrid (i.e., mobile biofilms and accumulated suspended biomass) bioreactor and liquid and solid separation. Mode II includes a mobile-biofilm retention screen that is downstream of and external to a hybrid bioreactor and upstream of liquid and solid separation. Mode III includes mobile-biofilm retention screening that is external to a hybrid bioreactor and liquid and solid separation, receives waste solids, and relies on environmental conditions and wastewater characteristics that are favorable for aerobic-granular sludge formation. This paper presents a mechanistic approach to design and evaluate MOB processes and describes MOB process: (1) modes of operation, (2) design and analysis methodology, (3) process and mechanical design criteria, (4) mathematical modeling, (5) design equations, and (6) mobile-biofilm settling characteristics and return. A mathematical model was applied to describe a fixed bioreactor volume and secondary-clarifier area with Modes I, II, and III. The mathematical modeling identified key differences between MOB process modes of operation, which are described in this paper. PRACTITIONER POINTS: MOB is a municipal and industrial wastewater treatment (WWT) process that reduces bioreactor and liquid and solids separation process volumes. It may operate with a mobile-biofilm reactor or a hybrid (mobile biofilms and suspended biomass) bioreactor. This paper provides a mechanistic basis for the selection and design of a MOB process mode of operation, and it describes MOB process modes of operation, design criteria, design equations, mathematical modeling, and mobile-biofilm settling characteristics. MOB integrated WWT plants exist at full scale and reliably meet their treatment objectives. The MOB process is an emerging environmental biotechnology for cost-effective WWT.

RevDate: 2022-09-16

Oğuz SK, Has EG, Akçelik N, et al (2022)

Phenotypic Impacts and Genetic Regulation Characteristics of the DNA Adenine Methylase Gene (dam) in S. Typhimurium Biofilm Forms.

Research in microbiology pii:S0923-2508(22)00072-9 [Epub ahead of print].

In this study, transcriptional level gene expression changes in biofilm forms of S. Typhimurium ATCC 14028 and its dam mutant were investigated by performing RNAseq analysis. As a result of these analyzes, a total of 233 differentially expressed genes (DEGs) were identified in the dam mutant, of which 145 genes were downregulated and 88 genes were upregulated compared to the wild type. According to data from miRNA sequence analysis, of 13 miRNAs differentially expressed in dam mutant, 9 miRNAs were downregulated and 4 miRNAs were upregulated. These data provide the first evidence that the dam gene is a global regulator of biofilm formation in Salmonella. In addition, phenotypic analyses revealed that bacterial swimming and swarming motility and cellulose production were highly inhibited in the dam mutant. It was determined that bacterial adhesion in Caco-2 and HEp-2 cell lines was significantly reduced in dam mutant. At the end of 90 minutes, the adhesion rate of wild type strain was 43.3% in Caco-2 cell line, while this rate was 14.9% in dam mutant. In the HEp-2 cell line, while 45.5% adherence was observed in the wild-type strain, this rate decreased to 15.3% in the dam mutant.

RevDate: 2022-09-16

Gong Y, Yin S, Sun S, et al (2022)

Chelerythrine reverses the drug resistance of resistant Candida albicans and the biofilm to fluconazole.

Future microbiology [Epub ahead of print].

Aim: To evaluate the antifungal activity of chelerythrine in combination with fluconazole against planktonic Candida albicans strains and preformed biofilm. Materials & methods: A broth microdilution assay was used to reveal the antifungal activity of chelerythrine combined with fluconazole against C. albicans and the preformed biofilm. A fractional inhibitory concentration index model was used to evaluate the interaction. Results: Chelerythrine strongly synergized with fluconazole against fluconazole-resistant C. albicans and the biofilm preformed for less than 12 h. In addition, chelerythrine combined with fluconazole exhibited a synergistic effect against C. albicans morphogenesis. Conclusion: Chelerythrine could reverse the drug resistance of resistant C. albicans and its biofilm to fluconazole, providing new insights for overcoming the drug resistance of C. albicans.

RevDate: 2022-09-17

Pawar RO, Narote PS, Gawai KT, et al (2022)

Comparative Analysis of Biofilm Formation on Materials Used for the Fabrication of Implant Supported Prostheses.

Journal of pharmacy & bioallied sciences, 14(Suppl 1):S812-S815.

Background: Zirconia and other dental pottery, heat-restored polymethyl methacrylate, titanium and other metal amalgams, or a mix of these materials are utilized to fix inserts. It is important to choose a material that is resistant to bacterial colonization for implant-supported prostheses, but durability and aesthetics are important factors as well.

Aim: Biofilm generation on materials used in implant-supported dental prosthesis manufacturing was an objective of this investigation.

Methods and Materials: In this study, 90 discs were prepared. These discs were divided into three groups: group PMMA, group Y-TZP, and group CP-Ti. Each group consisted of 30 discs. As helpful materials, 30 discs (D = 15 mm, H = 3 mm) each were created from either monetarily unadulterated titanium (CP-Ti), yttria tetragonal zirconia polycrystal (YTZP), or hotness-restored polymethyl methacrylate (PMMA). The examples were cleaned as per acknowledged practices. The non-contact profilometer (NPFLEX, Bruker, UK) was utilized to survey the surface rougness of each disc, and the outcomes were accounted for as Ra (m). An assortment of gram-negative microbes, including Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Porphyromonas gingivalis, Prevotella intermedia, and Tannerella albicans, were refined close by cleaned discs produced using heat-relieved PMMA, Y-TZP, or CP-Ti to see which type of biofilm is shaped best. CFU/mL was the unit of estimation (state framing units per milliliter).

Results: Y-TZP discs have a substantially higher Ra (349 41 m) than PMMA and CP-Ti discs. Some bacteria that have been associated with peri-mucositis and peri-implantitis may be less prevalent on Y-TZP or CP-Ti discs. Biofilm development must be considered while making implant-supported prostheses using Y-TZP and CP-Ti.

Conclusion: Y-TZP and CP-Ti are preferred materials for implant-supported prosthesis production because of biofilm development.

RevDate: 2022-09-17

Vaddamanu SK, Vyas R, Kavita K, et al (2022)

An In Vitro study to Compare Dental Laser with other Treatment Modalities on Biofilm Ablation from Implant and Tooth Surfaces.

Journal of pharmacy & bioallied sciences, 14(Suppl 1):S530-S533.

Background: Periodontal and peri-implant disorders are etiologically linked to bacterial biofilms. The researchers wanted to see how well the erbium-doped yttrium aluminum garnet (Er:YAG) laser removed bacterial biofilms along with attached epithelial cells (EC), gingival fibroblasts (GF), in addition to osteoblast-like cells (OC) dentin along with titanium surfaces compared to previous therapy methods.

Methodology: 3.5 days were spent growing bacterial biofilms on standardized dentin and also titanium samplings using a sand-blasted along with the acid-etched surface. Following that, the specimens were positioned into pockets that had been formed artificially. The following approaches were used to remove biofilm: (1) Er:YAG, (2) photodynamic therapy (PDT), and (3) curette (CUR) along with supplementary PDT (CUR/PDT). The remaining biofilms' colony forming units (CFUs) were determined, as well as the attachment of EC, GF, in addition to OC. Analysis of variance with a posthoc least significant difference was utilized in the statistical analysis.

Results: When compared to untreated dentin and titanium surfaces, all therapy strategies reduced total CFUs in statistically significant biofilms (p = 0.001). On the dentin, Er:YAG was as effective as CUR and PDT, but not as effective as CUR/PDT (p = 0.005). The application of Er:YAG on titanium surfaces leads to statistically significantly improved biofilm eradication equated to the supplementary three therapies (all p = 0.001). On untouched infested dentin and titanium surfaces, the counts of attached EC, GF, and OC were the lowermost. Atop the dentin, increased EC counts were detected after CUR/PDT (p = 0.006). On titanium, all cleaning procedures increased the counts of attached EC by a statistically significant amount (p = 0.001), with no variations between groups. After Er:YAG decontamination, there were statistically substantially elevated amounts of GF (p = 0.024) and OC (p = 0.001) than on untreated surfaces.

Conclusion: The usage of Er:YAG laser to ablate subgingival biofilms and, specifically, to decontaminate titanium implant surfaces appears to be a promising strategy that needs further research.

RevDate: 2022-09-17

Pawar M, Agwan MAS, Toshniwal NG, et al (2022)

Efficacy of Ozone to Eliminate Endopathogenic Microorganism in Rootcanal Biofilm.

Journal of pharmacy & bioallied sciences, 14(Suppl 1):S876-S879.

Objective: The researchers wanted to see whether ozonated water with ultrasonication and sodium hypochlorite can destroy Enterococcus faecalis bacteria in root canals.

Materials and Methods: A total of 40 single-rooted human teeth were used. A total of 100 roots were harvested and mechanically prepared. The root canals were randomly divided into four classes (n = 10) after being infected with E. faecalis for 24 h. Each sample's MTT value was calculated.

Conclusions: NaOCl and aqueous ozone provide antibacterial effects in in-vitro conditions in root canals.

RevDate: 2022-09-17

Zhang D, Shen J, Peng X, et al (2022)

Physiological changes and growth behavior of Corynebacterium glutamicum cells in biofilm.

Frontiers in microbiology, 13:983545.

Biofilm cells are well-known for their increased survival and metabolic capabilities and have been increasingly implemented in industrial and biotechnological processes. Corynebacterium glutamicum is one of the most widely used microorganisms in the fermentation industry. However, C. glutamicum biofilm has been rarely reported and little is known about its cellular basis. Here, the physiological changes and characteristics of C. glutamicum biofilm cells during long-term fermentation were studied for the first time. Results showed that the biofilm cells maintained stable metabolic activity and cell size was enlarged after repeated-batch of fermentation. Cell division was slowed, and chromosome content and cell proliferation efficiency were reduced during long-term fermentation. Compared to free cells, more biofilm cells were stained by the apoptosis indicator dyes Annexin V-FITC and propidium iodide (PI). Overall, these results suggested slow-growing, long-lived cells of C. glutamicum biofilm during fermentation, which could have important industrial implications. This study presents first insights into the physiological changes and growth behavior of C. glutamicum biofilm cell population, which would be valuable for understanding and developing biofilm-based processes.

RevDate: 2022-09-21
CmpDate: 2022-09-19

Tillander JAN, Rilby K, Svensson Malchau K, et al (2022)

Treatment of periprosthetic joint infections guided by minimum biofilm eradication concentration (MBEC) in addition to minimum inhibitory concentration (MIC): protocol for a prospective randomised clinical trial.

BMJ open, 12(9):e058168.

INTRODUCTION: Prosthetic joint infections (PJIs) are disastrous complications for patients and costly for healthcare organisations. They may promote bacterial resistance due to the extensive antibiotic use necessary in the PJI treatment. The PJI incidence is estimated to be 1%-3%, but the absolute numbers worldwide are high and increasing as large joint arthroplasties are performed by the millions each year. Current treatment algorithms, based on implant preserving surgery or full revision followed by a semitailored antibiotic regimen for no less than 2-3 months, lead to infection resolution in approximately 60% and 90%, respectively. Antibiotic choice is currently guided by minimum inhibitory concentrations (MICs) of free-living bacteria and not of bacteria in biofilm growth mode. Biofilm assays with relatively rapid output for the determination of minimum biofilm eradication concentrations (MBECs) have previously been developed but their clinical usefulness have not been established.

METHODS AND ANALYSIS: This single-blinded, two-arm randomised study of hip or knee staphylococcal PJI will evaluate 6-week standard of care (MIC guided), or an alternative antibiotic regimen according to an MBEC-guided-based decision algorithm. Sixty-four patients with a first-time PJI treated according to the debridement, antibiotics, and implant retention principle will be enrolled at a single tertiary orthopaedic centre (Sahlgrenska University Hospital). Patients will receive 14 days of standard parenteral antibiotics before entering the comparative study arms. The primary outcome measurement is the proportion of changes in antimicrobial regimen from first-line treatment dependent on randomisation arm. Secondary endpoints are unresolved infection, how microbial properties including biofilm abilities and emerging antimicrobial resistance correlate to infection outcomes, patient reported outcomes and costs with a 12-month follow-up.

ETHICS AND DISSEMINATION: Approval is received from the Swedish Ethical Review Authority, no 2020-01471 and the Swedish Medical Products Agency, EudraCT, no 2020-003444-80.

TRIAL REGISTRATION NUMBER: ClinicalTrials.gov ID: NCT04488458.

RevDate: 2022-09-15

Stepanov D, Buchmann D, Schultze N, et al (2022)

A Combined Bayesian and Similarity-Based Approach for Predicting E. coli Biofilm Inhibition by Phenolic Natural Compounds.

Journal of natural products [Epub ahead of print].

Screening for biofilm inhibition by purified natural compounds is difficult due to compounds' chemical diversity and limited commercial availability, combined with time- and cost-intensiveness of the laboratory process. In silico prediction of chemical and biological properties of molecules is a widely used technique when experimental data availability is of concern. At the same time, the performance of predictive models directly depends on the amount and quality of experimental data. Driven by the interest in developing a model for prediction of the antibiofilm effect of phenolic natural compounds such as flavonoids, we performed experimental assessment of antibiofilm activity of 320 compounds from this subset of chemicals. The assay was performed once on two Escherichia coli strains on agar in 24-well microtiter plates. The inhibition was assessed visually by detecting morphological changes in macrocolonies. Using the data obtained, we subsequently trained a Bayesian logistic regression model for prediction of biofilm inhibition, which was combined with a similarity-based method in order to increase the overall sensitivity (at the cost of accuracy). The quality of the predictions was subsequently validated by experimental assessment in three independent experiments with two resistant E. coli strains of 23 compounds absent in the initial data set. The validation demonstrated that the model may successfully predict the targeted effect as compared to the baseline accuracy. Using a randomly selected database of commercially available natural phenolics, we obtained approximately 6.0% of active compounds, whereas using our prediction-based substance selection, the percentage of phenolics found to be active increased to 34.8%.

RevDate: 2022-09-16
CmpDate: 2022-09-16

Rumbaugh KP (2022)

Dispersing biofilm myths.

Nature reviews. Microbiology, 20(10):573-574.

RevDate: 2022-09-16
CmpDate: 2022-09-16

Blaskovich MAT, Hansford KA, Butler MS, et al (2022)

A lipoglycopeptide antibiotic for Gram-positive biofilm-related infections.

Science translational medicine, 14(662):eabj2381.

Drug-resistant Gram-positive bacterial infections are still a substantial burden on the public health system, with two bacteria (Staphylococcus aureus and Streptococcus pneumoniae) accounting for over 1.5 million drug-resistant infections in the United States alone in 2017. In 2019, 250,000 deaths were attributed to these pathogens globally. We have developed a preclinical glycopeptide antibiotic, MCC5145, that has excellent potency (MIC90 ≤ 0.06 μg/ml) against hundreds of isolates of methicillin-resistant S. aureus (MRSA) and other Gram-positive bacteria, with a greater than 1000-fold margin over mammalian cell cytotoxicity values. The antibiotic has therapeutic in vivo efficacy when dosed subcutaneously in multiple murine models of established bacterial infections, including thigh infection with MRSA and blood septicemia with S. pneumoniae, as well as when dosed orally in an antibiotic-induced Clostridioides difficile infection model. MCC5145 exhibited reduced nephrotoxicity at microbiologically active doses in mice compared to vancomycin. MCC5145 also showed improved activity against biofilms compared to vancomycin, both in vitro and in vivo, and a low propensity to select for drug resistance. Characterization of drug action using a transposon library bioinformatic platform showed a mechanistic distinction from other glycopeptide antibiotics.

RevDate: 2022-09-16
CmpDate: 2022-09-16

Powell LC, Cullen JK, Boyle GM, et al (2022)

Topical, immunomodulatory epoxy-tiglianes induce biofilm disruption and healing in acute and chronic skin wounds.

Science translational medicine, 14(662):eabn3758.

The management of antibiotic-resistant, bacterial biofilm infections in chronic skin wounds is an increasing clinical challenge. Despite advances in diagnosis, many patients do not derive benefit from current anti-infective/antibiotic therapies. Here, we report a novel class of naturally occurring and semisynthetic epoxy-tiglianes, derived from the Queensland blushwood tree (Fontainea picrosperma), and demonstrate their antimicrobial activity (modifying bacterial growth and inducing biofilm disruption), with structure/activity relationships established against important human pathogens. In vitro, the lead candidate EBC-1013 stimulated protein kinase C (PKC)-dependent neutrophil reactive oxygen species (ROS) induction and NETosis and increased expression of wound healing-associated cytokines, chemokines, and antimicrobial peptides in keratinocytes and fibroblasts. In vivo, topical EBC-1013 induced rapid resolution of infection with increased matrix remodeling in acute thermal injuries in calves. In chronically infected diabetic mouse wounds, treatment induced cytokine/chemokine production, inflammatory cell recruitment, and complete healing (in six of seven wounds) with ordered keratinocyte differentiation. These results highlight a nonantibiotic approach involving contrasting, orthogonal mechanisms of action combining targeted biofilm disruption and innate immune induction in the treatment of chronic wounds.

RevDate: 2022-09-14

Dışhan A, Hizlisoy H, Barel M, et al (2022)

Biofilm formation, antibiotic resistance and genotyping of shiga toxin-producing Escherichia coli Isolated from retail chicken meats.

British poultry science [Epub ahead of print].

1. The shiga toxin-producing Escherichia coli (STEC) bacterium is a hazardous zoonotic agent for chicken meat consumers. This study determined the serogroups and evaluated the virulence genes, antibiotic resistance, biofilm-forming profiles and genetic relationships of STEC isolates in chicken meat.2. A total of 100 samples belonging to dressed-whole chicken and different parts of the chicken (wing, breast, thigh, drumstick) were collected between September and November 2019 from farms in Kayseri, Turkey.3. The phenotypic (identification, disc diffusion test, Congo red agar and microtitre plate tests) and molecular tests (identification, serogrouping, virulence factors, biofilm, antibiotic susceptibility, 16S rRNA sequencing and enterobacterial repetitive intergenic consensus-PCR for typing of the isolates) were carried out.4. E. coli was isolated from 35% of the samples and 35% of the samples harboured at least one STEC. Among 35 STEC isolates, 3 (8.5%), 6 (17.1%), 2 (5.7%) and 3 (8.5%) were found to be positive for fliCH2, fliCH8, fliCH11, fliCH19 genes, respectively. Out of 35 STEC positive isolates, 4 (11.4%) were identified as E. coli O157, from which 2 (5.7%) were E. coli O157:H7. E. coli O157 was detected in two (10%), one (5%), one (5%) of the thigh, drumstick and whole chicken samples, respectively.5. Biofilm-forming ability was reported in 33(94.2%) of 35 E. coli isolates, whilst the biofilm-associated genes detected among 35 STEC isolates included csgA (88.5%), fimH (88.5%), bcsA (85.7%) and agn43 (14.2%) and papC (8.5%). The STEC strains showed resistance against ampicillin (88.5%) and erythromycin (88.5%), following by tetracycline (74.2%) and gentamicin (25.7%). However, the distribution of isolates harbouring blaCMY, ere(A), tet(A) and aac(3)-IV antibiotic resistance genes was found to be 17.1%, 11.4%, 85.7% and 5.7%, respectively.6. ERIC-PCR showed that E. coli strains obtained from different parts and whole of chicken samples had genetic diversities. ERIC-PCR patterns grouped strains of 35 STEC into eight clusters designated A-H, with 73% similarity. Proper hygiene measures and staff training are essential for public health during poultry processing and in retail stores to control STEC.

RevDate: 2022-09-14

Wang Y, Li C, Wang J, et al (2022)

The Efficacy of Colistin Combined with Amikacin or Levofloxacin against Pseudomonas aeruginosa Biofilm Infection.

Microbiology spectrum [Epub ahead of print].

Pseudomonas aeruginosa (PA) biofilm infection is clinically prevalent and difficult to eradicate. In the present work, we aimed to evaluate the in vitro and in vivo efficacy of colistin (COL)-based combinations against PA biofilm. MICs and fractional inhibitory concentration indexes (FICIs) of four antibiotics (COL, amikacin, levofloxacin, and meropenem) to bioluminescent strain PAO1, carbapenem-resistant PAO1 (CRPAO1), and clinically isolated strains were assessed. Minimal biofilm eradication concentrations (MBECs) of monotherapy and combinations were examined by counting the live bacteria in biofilm, accompanied by visual confirmation using confocal laser-scanning microscopy. An animal biofilm infection model was established by implanting biofilm subcutaneously, and the therapeutic effect was evaluated according to the change in luminescence through a live animal bio-photonic imaging system. In vitro, even combined with 4 or 8 mg/L COL, meropenem needed to reach 128 or 256 mg/L to eradicate the biofilm. Moreover, 2 mg/L COL combined with 32 mg/L amikacin or 4-8 mg/L levofloxacin could kill the PAO1 and CRPAO1 in biofilm within 24 h. In vivo, COL combined with amikacin or levofloxacin could shorten the eradication time of biofilm than monotherapy. For PAO1 biofilm, combination therapy could eradicate the biofilm in all mice on the 5th day, whereas monotherapy only eradicated biofilms in almost half of the mice. For CRPAO1 biofilm, the biofilm eradication rate on the 6th day in the COL+ amikacin, amikacin, or COL alone regimen was 90%, 10%, or 40%, respectively. COL combined with levofloxacin did not show a better effect than each individual antibiotic. COL-based combinations containing levofloxacin or amikacin were promising choices for treating PA biofilm infection. IMPORTANCE Infections associated with PA biofilm formation are extremely challenging. When monotherapy fails to achieve optimal efficacy, combination therapy becomes the last option. After evaluating multiple drug combinations through a series of experiments in vitro and in vivo, we confirmed that colistin-based combinations containing levofloxacin or amikacin were promising choices for treating PA biofilm infection. The efficacy of these combinations derives from the different bactericidal mechanisms and the bacterial susceptibility to each antibiotic. This study provided a new regimen to solve the incurable problem of biofilm by using COL combined with other antibiotics.

RevDate: 2022-09-17

Ballah FM, Islam MS, Rana ML, et al (2022)

Phenotypic and Genotypic Detection of Biofilm-Forming Staphylococcus aureus from Different Food Sources in Bangladesh.

Biology, 11(7):.

Staphylococcus aureus is a major foodborne pathogen. The ability of S. aureus to produce biofilm is a significant virulence factor, triggering its persistence in hostile environments. In this study, we screened a total of 420 different food samples and human hand swabs to detect S. aureus and to determine their biofilm formation ability. Samples analyzed were meat, milk, eggs, fish, fast foods, and hand swabs. S. aureus were detected by culturing, staining, biochemical, and PCR. Biofilm formation ability was determined by Congo Red Agar (CRA) plate and Crystal Violet Microtiter Plate (CVMP) tests. The icaA, icaB, icaC, icaD, and bap genes involved in the synthesis of biofilm-forming intracellular adhesion compounds were detected by PCR. About 23.81% (100/420; 95% CI: 14.17-29.98%) of the samples harbored S. aureus, as revealed by detection of the nuc gene. The CRA plate test revealed 20% of S. aureus isolates as strong biofilm producers and 69% and 11% as intermediate and non-biofilm producers, respectively. By the CVMP staining method, 20%, 77%, and 3% of the isolates were found to be strong, intermediate, and non-biofilm producers. Furthermore, 21% of S. aureus isolates carried at least one biofilm-forming gene, where icaA, icaB, icaC, icaD, and bap genes were detected in 15%, 20%, 7%, 20%, and 10% of the S. aureus isolates, respectively. Bivariate analysis showed highly significant correlations (p < 0.001) between any of the two adhesion genes of S. aureus isolates. To the best of our knowledge, this is the first study in Bangladesh describing the detection of biofilm-forming S. aureus from foods and hand swabs using molecular-based evidence. Our findings suggest that food samples should be deemed a potential reservoir of biofilm-forming S. aureus, which indicates a potential public health significance.

RevDate: 2022-09-13

Akao PK, Kaplan A, Avisar D, et al (2022)

Removal of carbamazepine, venlafaxine and iohexol from wastewater effluent using coupled microalgal-bacterial biofilm.

Chemosphere pii:S0045-6535(22)02892-2 [Epub ahead of print].

We evaluated the removal capacity of a coupled microalgal-bacterial biofilm (CMBB) to eliminate three recalcitrant pharmaceuticals. The CMBB's efficiency, operating at different biofilm concentrations, with or without light, was compared and analyzed to correlate these parameters to pharmaceutical removal and their effect on the microorganism community. Removal rates changed with changing pharmaceutical and biofilm concentrations: higher biofilm concentrations presented higher removal. Removal of 82-94% venlafaxine and 18-51% carbamazepine was obtained with 5 days of CMBB treatment. No iohexol removal was observed. Light, microorganism composition, and dissolved oxygen concentration are essential parameters governing the removal of pharmaceuticals and ammonia. Chlorophyll concentration increased with time, even in the dark. Three bacterial phyla were dominant: Proteobacteria, Bacteroidetes and Firmicutes. The dominant eukaryotic supergroups were Archaeplastida, Excavata and SAR. A study of the microorganisms' community indicated that not only do the species in the biofilm play an important role; environment, concentration and interactions among them are also important. CMBB has the potential to provide low-cost and sustainable treatment for wastewater and recalcitrant pharmaceutical removal. The microenvironments on the biofilm created by the microalgae and bacteria improved treatment efficiency.

RevDate: 2022-09-13

Mohd Badri PEA, Rismayuddin NAR, Kenali NM, et al (2022)

Characterization of Cervus timorensis Velvet Antler and its Effect on Biofilm Formation of Candida Species.

Medical mycology pii:6696968 [Epub ahead of print].

Oral biofilms comprise extracellular polysaccharides and polymicrobial microorganisms. The objectives of the study were to characterize the deer velvet antler (DVA) compounds and their effect on Candida species biofilm formation with the hypothesis that DVA inhibits the biofilm of Candida spp. Liquid Chromatography-Quadrupole Time of Flight-Mass Spectrometry (LC-QTOF-MS) was conducted to characterize the DVA compounds. To study the effect of DVA on biofilm, Candida albicans ATCC MYA-4901 (ALT5), AIDS isolate (ALC2), oral cancer isolate (ALC3), C. dubliniensis ATCC MYA-2975, C. glabrata ATCC 90030, C. krusei 14 243, C. lusitaniae ATCC 34449, C. parapsilosis ATCC 22019, and C. tropicalis ATCC 13803 were inoculated with DVA in separate wells of a 96-well plate containing RPMI-1640 followed by 72 h incubation. A total of 45 compounds were detected in the DVA extract. C. lusitaniae exhibited a higher percentage of biofilm biomass reduction when treated with DVA extract (66.10% ± 5.33), followed by ALC3 (44.12% ± 6.24). However, C. glabrata, C. krusei, and C. parapsilosis showed no reduction in biofilm biomass after being treated with DVA extract. Most Candida strains also exhibited decreased total cell count when treated with DVA extract, except for ALC3 and C. krusei. ALT5 had the lowest total cell count (0.17 × 105 cells/mL) when cultured with DVA extract. In conclusion, DVA extract inhibits Candida spp. biofilm formation except for C. glabrata, C. krusei, and C. parapsilosis.

RevDate: 2022-09-13

Vanajothi R, Bhavaniramya S, Vijayakumar R, et al (2022)

In silico and In vitro Analysis of Nigella sativa Bioactives Against Chorismate Synthase of Listeria monocytogenes: a Target Protein for Biofilm Inhibition.

Applied biochemistry and biotechnology [Epub ahead of print].

Listeria monocytogenes have the ability to form biofilms, which aid in the contamination of food and the evasion of antimicrobials. Consumption of L. monocytogenes laden food can promote mild to severe infection in humans and cause serious health issues. Therefore, biofilm development by L. monocytogenes is considered to be a major concern for both healthcare and food safety. This study attempted to target chorismate synthase, an essential protein predicted to be involved in the biofilm pathway. Nigella sativa is renowned for its applications in folk medicine; hence, bioactive ingredients reported were used for molecular docking studies. In the absence of a three-dimensional structure of chorismate synthase from L. monocytogenes, a homology model was generated using the Modeller program. A model with the highest DOPE score was chosen and validated. The reliable model was subjected to docking studies with 30 ligands from N. sativa. From this approach, α-longipinene was unveiled as the best hit. Further in vitro studies demonstrated the antibiofilm potential of α-longipinene against L. monocytogenes. Overall, the study reveals lead molecules from N. sativa as promising antibiofilm agents against L. monocytogenes. Hence, extended investigation with lead molecules will provide sustainable strategies to prevent biofilm-mediated problems due to L. monocytogenes.


RJR Experience and Expertise


Robbins holds BS, MS, and PhD degrees in the life sciences. He served as a tenured faculty member in the Zoology and Biological Science departments at Michigan State University. He is currently exploring the intersection between genomics, microbial ecology, and biodiversity — an area that promises to transform our understanding of the biosphere.


Robbins has extensive experience in college-level education: At MSU he taught introductory biology, genetics, and population genetics. At JHU, he was an instructor for a special course on biological database design. At FHCRC, he team-taught a graduate-level course on the history of genetics. At Bellevue College he taught medical informatics.


Robbins has been involved in science administration at both the federal and the institutional levels. At NSF he was a program officer for database activities in the life sciences, at DOE he was a program officer for information infrastructure in the human genome project. At the Fred Hutchinson Cancer Research Center, he served as a vice president for fifteen years.


Robbins has been involved with information technology since writing his first Fortran program as a college student. At NSF he was the first program officer for database activities in the life sciences. At JHU he held an appointment in the CS department and served as director of the informatics core for the Genome Data Base. At the FHCRC he was VP for Information Technology.


While still at Michigan State, Robbins started his first publishing venture, founding a small company that addressed the short-run publishing needs of instructors in very large undergraduate classes. For more than 20 years, Robbins has been operating The Electronic Scholarly Publishing Project, a web site dedicated to the digital publishing of critical works in science, especially classical genetics.


Robbins is well-known for his speaking abilities and is often called upon to provide keynote or plenary addresses at international meetings. For example, in July, 2012, he gave a well-received keynote address at the Global Biodiversity Informatics Congress, sponsored by GBIF and held in Copenhagen. The slides from that talk can be seen HERE.


Robbins is a skilled meeting facilitator. He prefers a participatory approach, with part of the meeting involving dynamic breakout groups, created by the participants in real time: (1) individuals propose breakout groups; (2) everyone signs up for one (or more) groups; (3) the groups with the most interested parties then meet, with reports from each group presented and discussed in a subsequent plenary session.


Robbins has been engaged with photography and design since the 1960s, when he worked for a professional photography laboratory. He now prefers digital photography and tools for their precision and reproducibility. He designed his first web site more than 20 years ago and he personally designed and implemented this web site. He engages in graphic design as a hobby.

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An examination of the research and translational application to prevent and treat biofilm-associated diseases In the decade since the first edition of Microbial Biofilms was published, the interest in this field has expanded, spurring breakthrough research that has advanced the treatment of biofilm-associated diseases. This second edition takes the reader on an exciting, extensive review of bacterial and fungal biofilms, ranging from basic molecular interactions to innovative therapies, with particular emphasis on the division of labor in biofilms, new approaches to combat the threat of microbial biofilms, and how biofilms evade the host defense.

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Collection of publications by R J Robbins

Reprints and preprints of publications, slide presentations, instructional materials, and data compilations written or prepared by Robert Robbins. Most papers deal with computational biology, genome informatics, using information technology to support biomedical research, and related matters.

Research Gate page for R J Robbins

ResearchGate is a social networking site for scientists and researchers to share papers, ask and answer questions, and find collaborators. According to a study by Nature and an article in Times Higher Education , it is the largest academic social network in terms of active users.

Curriculum Vitae for R J Robbins

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Curriculum Vitae for R J Robbins

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RJR Picks from Around the Web (updated 11 MAY 2018 )