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Anti-Biofilm Activity of a Low Weight Proteinaceous Molecule from the Marine Bacterium Pseudoalteromonas sp. IIIA004 against Marine Bacteria and Human Pathogen Biofilms
Florie Desriac
Microorganisms
Pseudoalteromonas bacteria are known as potential bioactive metabolite producers. Because of the need to obtain natural molecules inhibiting the bacterial biofilms, we investigated the biofilm inhibitory activity of the marine bacterium Pseudoalteromonas sp. IIIA004 against the pioneer surface colonizer Roseovarius sp. VA014. The anti-biofilm activity from the culture supernatant of Pseudoalteromonas sp. IIIA004 (SNIIIA004) was characterized in microtiter plates (static conditions/polystyrene surface) and in flow cell chambers (dynamic conditions/glass surface). The Pseudoalteromonas exoproducts exhibited an inhibition of Roseovarius sp. VA014 biofilm formation as well as a strong biofilm dispersion, without affecting the bacterial growth. Microbial adhesion to solvent assays showed that SNIIIA004 did not change the broad hydrophilic and acid character of the Roseovarius strain surface. Bioassay-guided purification using solid-phase extraction and C18 reverse-phase-high-performance ...
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Antibiofilm Activity of the Marine Bacterium Pseudoalteromonas sp. Strain 3J6
Sophie Rodrigues, Chantal Compere
Applied and Environmental Microbiology, 2010
Biofilm formation results in medical threats or economic losses and is therefore a major concern in a variety of domains. In two-species biofilms of marine bacteria grown under dynamic conditions, Pseudoalteromonas sp. strain 3J6 formed mixed biofilms with Bacillus sp. strain 4J6 but was largely predominant over Paracoccus sp. strain 4M6 and Vibrio sp. strain D01. The supernatant of Pseudoalteromonas sp. 3J6 liquid culture (SN 3J6 ) was devoid of antibacterial activity against free-living Paracoccus sp. 4M6 and Vibrio sp. D01 cells, but it impaired their ability to grow as single-species biofilms and led to higher percentages of nonviable cells in 48-h biofilms. Antibiofilm molecules of SN 3J6 were able to coat the glass surfaces used to grow biofilms and reduced bacterial attachment about 2-fold, which might partly explain the biofilm formation defect but not the loss of cell viability. SN 3J6 had a wide spectrum of activity since it affected all Gram-negative marine strains tested except other Pseudoalteromonas strains. Biofilm biovolumes of the sensitive strains were reduced 3-to 530-fold, and the percentages of nonviable cells were increased 3-to 225-fold. Interestingly, SN 3J6 also impaired biofilm formation by three strains belonging to the human-pathogenic species Pseudomonas aeruginosa, Salmonella enterica, and Escherichia coli. Such an antibiofilm activity is original and opens up a variety of applications for Pseudoalteromonas sp. 3J6 and/or its active exoproducts in biofilm prevention strategies.
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Unraveling the anti-biofilm potential of green algal sulfated polysaccharides against Salmonella enterica and Vibrio harveyi
Jyoti Vishwakarma
Applied Microbiology and Biotechnology, 2020
One of the main reasons for the bacterial resistance to antibiotics is caused by biofilm formation of microbial pathogens during bacterial infections. Salmonella enterica and Vibrio harveyi are known to form biofilms and represent a major health concern worldwide, causing human infections responsible for morbidity and mortality. The current study aims to investigate the effect of purified sulfated polysaccharides (SPs) from Chlamydomonas reinhardtii (Cr) on planktonic and biofilm growth of these bacteria. The effect of Cr-SPs on bacterial planktonic growth was assessed by using the agar well diffusion method, which showed clear zones ranging from 13 to 26 mm in diameter from 0.5 to 8 mg/mL of Cr-SPs against both the bacteria. Time-kill activity and reduction in clonogenic propagation further help to understand the anti-microbial potential of Cr-SPs. The minimum inhibitory concentration of Cr-SPs against S. enterica and V. harveyi was as low as 440 μg/mL and 490 μg/mL respectively. Cr-SPs inhibited bacterial cell attachment up to 34.65-100% at 0.5-8 mg/mL in S. enterica and V. harveyi respectively. Cr-SPs also showed 2-fold decrease in the cell surface hydrophobicity, indicating their potential to prevent bacterial adherence. Interestingly, Cr-SPs efficiently eradicated the preformed biofilms. Increased reduction in total extracellular polysaccharide (EPS) and extracellular DNA (eDNA) content in a dose-dependent manner demonstrates Cr-SPs ability to interact and destroy the bacterial EPS layer. SEM analysis showed that Cr-SPs effectively distorted preformed biofilms and also induced morphological changes. Furthermore, Cr-SPs also showed anti-quorum-sensing potential by reducing bacterial urease and protease activities. These results indicate the potential of Cr-SPs as an anti-biofilm agent and will help to develop them as alternative therapeutics against biofilm-forming bacterial infections. Key points • Cr-SPs not only inhibited biofilm formation but also eradicated preformed biofilms. • Cr-SPs altered bacterial cell surface hydrophobicity preventing biofilm formation. • Cr-SPs efficiently degraded eDNA of the EPS layer disrupting mature biofilms. • Cr-SPs reduced activity of quorum-sensing-mediated enzymes like protease and urease.
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On the influence of the culture conditions in bacterial antifouling bioassays and biofilm properties: Shewanella algae, a case study
Roberto Dorta-Guerra
BMC Microbiology, 2014
Background: A variety of conditions (culture media, inocula, incubation temperatures) are employed in antifouling tests with marine bacteria. Shewanella algae was selected as model organism to evaluate the effect of these parameters on: bacterial growth, biofilm formation, the activity of model antifoulants, and the development and nanomechanical properties of the biofilms. The main objectives were: 1) To highlight and quantify the effect of these conditions on relevant parameters for antifouling studies: biofilm morphology, thickness, roughness, surface coverage, elasticity and adhesion forces. 2) To establish and characterise in detail a biofilm model with a relevant marine strain. Results: Both the medium and the temperature significantly influenced the total cell densities and biofilm biomasses in 24-hour cultures. Likewise, the IC 50 of three antifouling standards (TBTO, tralopyril and zinc pyrithione) was significantly affected by the medium and the initial cell density. Four media (Marine Broth, MB; 2% NaCl Mueller-Hinton Broth, MH2; Luria Marine Broth, LMB; and Supplemented Artificial Seawater, SASW) were selected to explore their effect on the morphological and nanomechanical properties of 24-h biofilms. Two biofilm growth patterns were observed: a clear trend to vertical development, with varying thickness and surface coverage in MB, LMB and SASW, and a horizontal, relatively thin film in MH2. The Atomic Force Microscopy analysis showed the lowest Young modulii for MB (0.16 ± 0.10 MPa), followed by SASW (0.19 ± 0.09 MPa), LMB (0.22 ± 0.13 MPa) and MH2 (0.34 ± 0.16 MPa). Adhesion forces followed an inverted trend, being higher in MB (1.33 ± 0.38 nN) and lower in MH2 (0.73 ± 0.29 nN).
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Bioactive Compounds of Pseudoalteromonas sp. IBRL PD4.8 Inhibit Growth of Fouling Bacteria and Attenuate Biofilms of Vibrio alginolyticus FB3
Nor Afifah Supardy
Polish Journal of Microbiology, 2019
Biofouling is a phenomenon that describes the fouling organisms attached to man-made surfaces immersed in water over a period of time. It has emerged as a chronic problem to the oceanic industries, especially the shipping and aquaculture fields. The metal-containing coatings that have been used for many years to prevent and destroy biofouling are damaging to the ocean and many organisms. Therefore, this calls for the critical need of natural product-based antifoulants as a substitute for its toxic counterparts. In this study, the antibacterial and antibiofilm activities of the bioactive compounds of Pseudoalteromonas sp. IBRL PD4.8 have been investigated against selected fouling bacteria. The crude extract has shown strong antibacterial activity against five fouling bacteria, with inhibition zones ranging from 9.8 to 13.7 mm and minimal inhibitory concentrations of 0.13 to 8.0 mg/ml. Meanwhile, the antibiofilm study has indicated that the extract has attenuated the initial and pre-f...
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Antibacterial Properties of Fucoidans from the Brown Algae Fucus vesiculosus L. of the Barents Sea
Elena Zhurishkina
Biology, 2021
Fucoidans, sulfated polysaccharides found in cell walls of brown algae, are considered as a promising antimicrobial component for various applications in medicine and the food industry. In this study, we compare the antibacterial properties of two fractions of fucoidan from the brown algae Fucus vesiculosus gathered in the littoral of the Barents Sea and sampled at different stages of purification. The crude fraction of fucoidan was isolated from algae by extraction with aqueous ethanol and sonication. The purified fraction was obtained by additional treatment of the crude fraction with a solution of calcium chloride. The structural features of both fractions were characterized in detail and their antibacterial effects against several Gram-positive and Gram-negative bacteria were compared by photometry, acridine orange staining assay, and atomic force microscopy. Fucoidan inhibited growth in all of the above microorganisms, showing a bacteriostatic effect with minimum inhibitory con...
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Evaluation of Anti-oxidant and Anti-biofilm Activities of Biogenic Surfactants Derived from Bacillus amyloliquefaciens and Pseudomonas aeruginosa
hossein rezvanifar
Iranian Journal of Pharmaceutical Research : IJPR, 2020
Biosurfactants, the microbial originated surface active agents, can modify the physicochemical properties of surfaces and reduce the bacterial adhesion via changing bacterial adhesion interactions on surfaces. They were also able to block oxidative chain reactions and might show antioxidant properties. The goal of this study was to evaluate the antioxidant and antibiofilm activities of biosurfactants which were derived from two autochthonous biosurfactant-producing strains, Bacillus amyloliquefaciens NS6 (surfactin), and Pseudomonas aeruginosa MN1 (rhamnolipids). Their antioxidant activities were determined by ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) methods. Ferric thiocyanate (FTC) assay was used for determination of their lipid peroxidation inhibition capacity. Their effect to reduce the adhesion of Streptococcus mutans on polystyrene surfaces and disruption of its pre-formed biofilms were also investigated. Our results indicated that surf...
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Evaluating the antibacterial and antibiofilm potential of sulphated polysaccharides extracted from green algaeChlamydomonas reinhardtii
Jyoti Vishwakarma
Journal of Applied Microbiology, 2019
Aim of the study: In the current study, antibacterial and antibiofilm potential of sulfated polysaccharides (SPs) extracted from Chlamydomonas reinhardtii (Cr) was evaluated against N. mucosa, E. coli, Streptococcus spp and B. subtilis. Methods and Results: Antibacterial potential of Cr-SPs was evaluated by agar-cup diffusion, time-kill and colony forming ability (CFU), minimum inhibitory and bactericidal concentration assays. Antibiofilm potential was evaluated by biofilm inhibition, eradication, extracellular-DNA, metabolic activity and microscopy assays. Cr-SPs at 0.5 mg/ml showed 34.52%, 48.6%, 66.1% and 55.6% reduced CFU in B. subtilis, Streptococcus, N. mucosa, E. coli respectively. MIC of Cr-SPs was as low as 480µg/ml for Streptococcus, N. mucosa and 420µg/ml for B. subtilis and E. coli. At 1mg/ml Cr-SPs showed 50% biofilm inhibition, while 4-8 mg/ml showed 100% inhibition. Cr-SPs also effectively dissolved preformed biofilms. Dose-dependent reduction in extracellular DNA revealed that Cr-SPs interacts with the extra polymeric substance of the biofilm and destroys them. Light microscopy reconfirmed the above results. Conclusion: Cr-SPs not only inhibited biofilm formation while effectively dissolved preformed-biofilms. Significance and Impact of the study: The current study showed the promising potential of Cr-SPs as antibiofilm agents. Further validation will help in developing Cr-SPs as natural antibiotics against biofilm causing bacteria.
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Anti-biofilm activity of the Antarctic marine bacterium Pseudoalteromonas haloplanktis TAC125
Ermenegilda Parrilli
Research in Microbiology, 2013
Considering the increasing impact of bacterial biofilms on human health, industrial and food-processing activities, the interest in the development of new approaches for the prevention and treatment of adhesion and biofilm formation capabilities has increased. A viable approach should target adhesive properties without affecting bacterial vitality in order to avoid the rapid appearance of escape mutants.
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Marine Biofilms as Mediators of Colonization by Marine Macroorganisms: Implications for Antifouling and Aquaculture
Sergey Dobretsov
Marine Biotechnology, 2007
In the marine environment, biofilms on submerged surfaces can promote or discourage the settlement of invertebrate larvae and macroalgal spores. The settlement-mediating effects of biofilms are believed to involve a variety of biofilm attributes including surface chemistry, micro-topography, and a wide range of microbial products from smallmolecule metabolites to high-molecular weight extracellular polymers. The settled organisms in turn can modify microbial species composition of biofilms and thus change the biofilm properties and dynamics. A better understanding of biofilm dynamics and chemical signals released and/or stored by biofilms will facilitate the development of antifouling and mariculture technologies. This review provides a brief account of 1) existing knowledge of marine biofilms that are relevant to settlement mediation, 2) biotechnological application of biofilms with respect to developing non-toxic antifouling technologies and improving the operation of aquaculture facilities, and 3) challenges and future directions for advancing our understanding of settlement-mediating functions of biofilms and for applying this knowledge to real-life situations.
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