• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.56-63
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• 2022

This study aims to determine the antibiofilm effect of cell-free supernatant (CFS) of Lactobacillus brevis strains against Streptococcus mutans strains. To study the antibiofilm mechanism against S. mutans strains, antibacterial effects, cell surface properties (auto-aggregation and cell surface hydrophobicity), exopolysaccharide (EPS) production, and morphological changes were examined. The antibiofilm effect of L. brevis KCCM 202399 CFS as morphological changes were evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), compared with the control treatment. Among the L. brevis strains, L. brevis KCCM 202399 showed the highest antibiofilm effect on S. mutans KCTC 5458. The antibacterial effect of L. brevis KCCM 202399 against S. mutans KCTC 5458 was investigated using the deferred method (16.00 mm). The minimum inhibitory concentration of L. brevis KCCM 202399 against S. mutans KCTC 5458 was 25.00%. Compared with the control treatment, L. brevis KCCM 202399 CFS inhibited the bacterial adhesion of S. mutans KCTC 5458 by decreasing auto-aggregation, cell surface hydrophobicity, and EPS production (45.91%, 40.51%, and 67.44%, respectively). L. brevis KCCM 202399 CFS inhibited and eradicated the S. mutans KCTC 5458 biofilm. Therefore, these results suggest that L. brevis KCCM 202399 CFS may be used to develop oral health in the probiotic industry.

• Microbiology and Biotechnology Letters
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• v.50 no.3
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• pp.328-336
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• 2022

Aptamers are short, chemically synthesized, single-stranded DNA or RNA oligonucleotides that fold into unique three-dimensional structures. In this study, we aim to determine the antibiofilm activity and binding specificity of the six polyclonal DNA aptamers (S15K3, S15K4, S15K6, S15K13, S15K15, and S15K20) on Staphylococcus aureus BPA-12 and Escherichia coli EPEC 4. Aptamer S15K6 showed the highest percentage of antibiofilm activity against S. aureus BPA-12 (37.4%) as shown by the lowest OD570 value of 0.313. Aptamer S15K20 showed the highest percentage of antibiofilm activity against E. coli EPEC 4 (15.4%) as shown by the lowest OD570 value of 0.515. Aptamers S15K13 and S15K20 showed antibiofilm activities against both S. aureus BPA-12 and E. coli EPEC4, and thus potentially have broad reactivity. Furthermore, based on the binding capacity and Kd values from our previous study, the binding specificity assay of selected polyclonal DNA aptamers (S15K3 and S15K15) against S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, S. agalactiae, E. coli MHA-6, and Listeria monocytogenes were performed using qPCR. Aptamers S15K3 and S15K15 showed specific binding to S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, and S. agalactiae, but could not bind to E. coli MHA-6 and L. monocytogenes. Therefore, this study showed that the polyclonal DNA aptamers have antibiofilm activity and were able to bind to S. aureus BPA-12 and E. coli EPEC 4 bacteria.

• Journal of Veterinary Science
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• v.25 no.2
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• pp.30.1-30.16
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• 2024

Background: Biofilms, such as those from Staphylococcus epidermidis, are generally insensitive to traditional antimicrobial agents, making it difficult to inhibit their formation. Although quercetin has excellent antibiofilm effects, its clinical applications are limited by the lack of sustained and targeted release at the site of S. epidermidis infection. Objectives: Polyethylene glycol-quercetin nanoparticles (PQ-NPs)-loaded gelatin-N,O-carboxymethyl chitosan (N,O-CMCS) composite nanogels were prepared and assessed for the on-demand release potential for reducing S. epidermidis biofilm formation. Methods: The formation mechanism, physicochemical characterization, and antibiofilm activity of PQ-nanogels against S. epidermidis were studied. Results: Physicochemical characterization confirmed that PQ-nanogels had been prepared by the electrostatic interactions between gelatin and N,O-CMCS with sodium tripolyphosphate. The PQ-nanogels exhibited obvious pH and gelatinase-responsive to achieve on-demand release in the micro-environment (pH 5.5 and gelatinase) of S. epidermidis. In addition, PQ-nanogels had excellent antibiofilm activity, and the potential antibiofilm mechanism may enhance its antibiofilm activity by reducing its relative biofilm formation, surface hydrophobicity, exopolysaccharides production, and eDNA production. Conclusions: This study will guide the development of the dual responsiveness (pH and gelatinase) of nanogels to achieve on-demand release for reducing S. epidermidis biofilm formation.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.542-551
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• 2017

Small phytochemicals have been successfully adopted as antibacterial chemotherapies and are being increasingly viewed as potential antibiofilm agents. Some of these molecules are known to repress biofilm and toxin production by certain bacterial and yeast pathogens, but information is lacking with regard to the genes allied with biofilm formation. The present study was performed to investigate the inhibitory effect of burdock root extract (BRE) and of chlorogenic acid (CGA; a component of BRE) on clinical isolates of Klebsiella pneumoniae. BRE and CGA exhibited significant antibiofilm activity against K. pneumoniae without inflicting any harm to its planktonic counterparts. In vitro assays supported the ${\beta}$-lactamase inhibitory effect of CGA and BRE while in silico docking showed that CGA bound strongly with the active sites of sulfhydryl-variable-1 ${\beta}$-lactamase. Furthermore, the mRNA transcript levels of two biofilm-associated genes (type 3 fimbriae mrkD and trehalose-6-phosphate hydrolase treC) were significantly downregulated in CGA- and BRE-treated samples. In addition, CGA inhibited biofilm formation by Escherichia coli and Candida albicans without affecting their planktonic cell growth. These findings show that BRE and its component CGA have potential use in antibiofilm strategies against persistent K. pneumoniae infections.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.193-201
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• 2022

Methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) are major causes of hospital- and community-acquired infections. The treatment of biofilm-related infections caused by these bacteria is a global healthcare challenge. Therefore, the development of alternative therapeutics is required. An essential oil extracted from Curcuma zedoaria (CZ) Rosc, also known as white turmeric, has been reported to possess various antimicrobial activities. In the present study, we evaluated the antibiofilm activities of an ethanolic extract of the CZ rhizome against MRSA and MSSA. The results showed that the CZ extract with the highest sub-minimum inhibitory concentration (sub-MIC), 1/2 MIC (0.312 mg/ml), significantly inhibited biofilm production by up to 80-90% in both tested strains. Subsequently, we evaluated the ability of the CZ extract to prevent cell-surface attachment to a 96-well plate and extracellular DNA (eDNA) release from the biofilm. The CZ extract demonstrated an inhibitory effect on bacterial attachment and eDNA release from the biofilm biomass. The CZ extract may inhibit biofilm formation by preventing eDNA release and cell-surface attachment. Therefore, this CZ extract is a potential candidate for the development of alternative treatments for biofilm-associated MRSA and MSSA infections.

• Korean Journal of Veterinary Research
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• v.61 no.2
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• pp.16.1-16.6
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• 2021

Pseudomonas aeruginosa is one of the most common pathogenic species associated with canine otitis externa (OE). Their resilience is achieved by forming a biofilm, which allows these bacteria to evade even the harshest of treatments. This study evaluated the in vitro synergistic efficacy of N-acetylcysteine (NAC) with different antimicrobial agents against P. aeruginosa isolated from dogs with OE to develop an effective treatment against P. aeruginosa. The antimicrobial activity was evaluated by the minimum inhibitory concentration test using the microdilution method. The efficacy of antibiofilm formation was evaluated using a crystal violet stain method. The treatment solutions included NAC alone, and in synergy with enrofloxacin, polymyxin B, and gentamicin. NAC alone exhibited antimicrobial and antibiofilm abilities. On the other hand, the combination of NAC and the antibiotics did not show any significant synergistic effects against P. aeruginosa.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1654-1663
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• 2018

Finding a safe and broad-spectrum medication is a goal of scientists, pharmacists, and physicians, but developing and fabricating the right medicine can be challenging. The current study describes the formation of silver nanoparticles (AgNPs) by Fusarium mangiferae. It involves the antibiofilm activity of the nanoparticles against Staphylococcus aureus. It also involves cytotoxic effect against mammalian cell lines. Well-dispersed nanoparticles are formed by F. mangiferae. The sizes of the nanoparticles were found to range from 25 to 52 nm, and UV-Vis scan showed absorption around 416-420 nm. SEM, TEM, and AFM results displayed spherical and oval shapes. Furthermore, the FTIR histogram detected amide I and amide II compounds responsible for the stability of AgNPs in an aqueous solution. AgNPs were observed to decrease the formation of biofilm at 75% (v/v). DNA reducing, smearing, and perhaps fragmentation were noticed after treating the bacterial cells with 50% (v/v). Additionally, cell lysis was detected releasing proteins in the supernatant. It was also observed that the AgNPs have the ability to cause 59% cervical cancer cell line (HeLa) deaths at 25% (v/v), however, they showed about 31% toxicity against rat embryo fibroblast transformed cell lines (REF). The results of this study prove the efficiency of AgNPs as an antibiofilm against S. aureus, suggesting that AgNPs could be an alternative to antibiotics. It must also be emphasized that AgNPs displayed cytotoxic behavior against mammalian cell lines. Further studies are needed for assessing risk in relation to the possible benefit of prescribing AgNPs.

• Natural Product Sciences
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• v.25 no.2
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• pp.172-180
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• 2019

Infections with Pseudomonas aeruginosa are difficult to treat not only because it is often associated with multidrug-resistant infections but also it is able to form biofilm. The aim of this study was to evaluate the antibiofilm and anti-Quorum Sensing (QS) activities of Thymbra capitata essential oils (EOs) against Beta Lactamase (BL) producing P. aeruginosa and the reference strain P. aeruginosa 10145. GC/MS analysis showed that thymol (23.25%) is the most dominant compound in T. capitata EOs. The MICs of T. capitata EOs against P. aeruginosa (BL) and P. aeruginosa 10145 were 1.11%. At sub MIC (0.041, 0.014 and 0.0046%), the EOs of T. capitata remarkably inhibited the biofilm formation of both strains tested and complete inhibition of the biofilm formation was reported at 0.041%. The EOs of T. capitata were found to inhibit the swarming motility, aggregation ability and hydrophobic ability of P. aeruginosa (BL) and P. aeruginosa 10145. Interestingly, the EOs of T. capitata reduce the production of three secreted virulence factors that regulated by QS system including pyocyanin, rhamnolipids and LasA protease. The potent antibiofilm and anti-QS activities of T. capitata EOs can propose it as a new antibacterial agent to control pseudomonas infections.

• Journal of Periodontal and Implant Science
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• v.48 no.6
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• pp.373-382
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• 2018

Purpose: The aim of this study was to evaluate the effects of various prophylactic treatments of titanium implants on bacterial biofilm formation, correlating surface modifications with the biofilms produced by Pseudomonas aeruginosa PAO1, Staphylococcus aureus, and bacteria isolated from saliva. Methods: Pure titanium disks were treated with various prophylactic procedures, and atomic force microscopy (AFM) was used to determine the degree to which surface roughness was modified. To evaluate antibiofilm activity, we used P. aeruginosa PAO1, S. aureus, and saliva-isolated Streptococcus spp., Bacteroides fragilis, and Staphylococcus epidermidis. Results: AFM showed that the surface roughness increased after using the air-polishing device and ultrasonic scaler, while a significant reduction was observed after using a curette or polishing with Detartrine ZTM (DZ) abrasive paste. In addition, we only observed a significant (P<0.01) reduction in biofilm formation on the DZ-treated implant surfaces. Conclusion: In this study, both AFM and antibiofilm analyses indicated that using DZ abrasive paste could be considered as the prophylactic procedure of choice for managing peri-implant lesions and for therapy-resistant cases of periodontitis.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.130-136
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• 2021

Persister cell formation and biofilms of pathogens are extensively involved in the development of chronic infectious diseases. Eradicating persister cells is challenging, owing to their tolerance to conventional antibiotics, which cannot kill cells in a metabolically dormant state. A high frequency of persisters in biofilms makes inactivating biofilm cells more difficult, because the biofilm matrix inhibits antibiotic penetration. Fatty acids may be promising candidates as antipersister or antibiofilm agents, because some fatty acids exhibit antimicrobial effects. We previously reported that fatty acid ethyl esters effectively inhibit Escherichia coli persister formation by regulating an antitoxin. In this study, we screened a fatty acid library consisting of 65 different fatty acid molecules for altered persister formation. We found that undecanoic acid, lauric acid, and N-tridecanoic acid inhibited E. coli BW25113 persister cell formation by 25-, 58-, and 44-fold, respectively. Similarly, these fatty acids repressed persisters of enterohemorrhagic E. coli EDL933. These fatty acids were all medium-chain saturated forms. Furthermore, the fatty acids repressed Enterohemorrhagic E. coli (EHEC) biofilm formation (for example, by 8-fold for lauric acid) without having antimicrobial activity. This study demonstrates that medium-chain saturated fatty acids can serve as antipersister and antibiofilm agents that may be applied to treat bacterial infections.