• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.348-355
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• 2023

Epifluorescence microscopy with image analysis was evaluated as a biofilm quantification method (i.e., quantification of surface area colonized by biofilms), in comparison with crystal violet (CV) staining. We performed different experiments to generate multispecies biofilms with natural and artificial bacterial assemblages. First, four species were inoculated daily in 16 different sequences to form biofilms (surface colonization, 0.1%-56.6%). Second, a 9-species assemblage was allowed to form biofilms under 10 acylase treatment episodes (33.8%-55.6%). The two methods comparably measured the quantitative variation in biofilms, exhibiting a strong positive relationship (R² ≥ 0.7). Moreover, the two methods exhibited similar levels of variation coefficients. Finally, six synthetic and two natural consortia were allowed to form biofilms for 14 days, and their temporal dynamics were monitored. The two methods were comparable in quantifying four biofilms colonizing ≥18.7% (R² ≥ 0.64), but not for the other biofilms colonizing ≤ 3.7% (R² ≤ 0.25). In addition, the two methods exhibited comparable coefficients of variation in the four biofilms. Microscopy and CV staining comparably measured the quantitative variation of biofilms, exhibiting a strongly positive relationship, although microscopy cannot appropriately quantify the biofilms below the threshold colonization. Microscopy with image analysis is a promising approach for easily and rapidly estimating absolute quantity of multispecies biofilms.

• Journal of Microbiology and Biotechnology
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• v.27 no.6
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• pp.1053-1064
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• 2017

A biofilm is a community of microbes that typically inhabit on surfaces and are encased in an extracellular matrix. Biofilms display very dissimilar characteristics to their planktonic counterparts. Biofilms are ubiquitous in the environment and influence our lives tremendously in both positive and negative ways. Pseudomonas aeruginosa is a bacterium known to produce robust biofilms. P. aeruginosa biofilms cause severe problems in immunocompromised patients, including those with cystic fibrosis or wound infection. Moreover, the unique biofilm properties further complicate the eradication of the biofilm infection, leading to the development of chronic infections. In this review, we discuss the history of biofilm research and general characteristics of bacterial biofilms. Then, distinct features pertaining to each stage of P. aeruginosa biofilm development are highlighted. Furthermore, infections caused by biofilms on their own or in association with other bacterial species (i.e., multispecies biofilms) are discussed in detail.

• Journal of Korean Society on Water Environment
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• v.23 no.1
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• pp.1-11
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• 2007

The low nutrient environment in drinking water treatment plants and distribution systems cannot to be a good environment for bacterial growth. However, biofilms can be frequently found submerged surface in treatment plants and distribution system. Biofilms in distribution system are harmful, in that they can release organisms, and may cause problems in taste and odor of water. Control of these Biofilms is difficult, and disinfection alone is usually ineffective. Biofilms will not be eliminated from distribution systems by any contemporary technology available now or in the future. Therefore reduction of organic matter, improved disinfection, or a combination of these methods can be useful in controlling distribution system biofilms.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.1-7
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• 2021

Bacterial biofilm is a community of bacteria that are embedded and structured in a self-secreted extracellular matrix. An important clinical-related characteristic of bacterial biofilms is that they are much more resistant to antimicrobial agents than the planktonic cells (up to 1,000 times), which is one of the main causes of antibiotic resistance in clinics. Therefore, infections caused by biofilms are notoriously difficult to eradicate, such as lung infection caused by Pseudomonas aeruginosa in cystic fibrosis patients. Understanding the resistance mechanisms of biofilms will provide direct insights into how we overcome such resistance. In this review, we summarize the characteristics of biofilms and chronic infections associated with bacterial biofilms. We examine the current understanding and research progress on the major mechanisms of antibiotic resistance in biofilms, including quorum sensing. We also discuss the potential strategies that may overcome biofilm-related antibiotic resistance, focusing on targeting biofilm EPSs, blocking quorum sensing signaling, and using recombinant phages.

• Geomechanics and Engineering
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• v.12 no.5
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• pp.863-870
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• 2017

With the increasing interest in using bacterial biofilms in geo-engineering practices, such as soil improvement, sealing leakage in earth structures, and hydraulic barrier installation, understanding of the contribution of bacterial biofilm formation to mechanical and hydraulic behavior of soils is important. While mechanical properties of soft gel-like biofilms need to be identified for appropriate modeling and prediction of behaviors of biofilm-associated soils, elastic properties of biofilms remain poorly understood. Therefore, this study investigated the microscale Young's modulus of biofilms produced by Shewanella oneidensis MR-1 in a liquid phase. The indentation test was performed on a biofilm sample using the atomic force microscopy (AFM) with a spherical indentor, and the force-indentation responses were obtained during approach and retraction traces. Young's modulus of biofilms was estimated to be ~33-38 kPa from these force-indentation curves and Hertzian contact theory. It appears that the AFM indentation result captures the microscale local characteristics of biofilms and its stiffness is relatively large compared to the other methods, including rheometer and hydrodynamic shear tests, which reflect the average macro-scale behaviors. While modeling of mechanical behaviors of biofilm-associated soils requires the properties of each component, the obtained results provide information on the mechanical properties of biofilms that can be considered as cementing, gluing, or filling materials in soils.

• Journal of Korean Academy of Oral Health
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• v.43 no.3
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• pp.118-123
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• 2019

Objectives: The purpose of this study was to investigate changes in the composition of artificial cariogenic biofilms using a Streptococcus mutans biofilm model over a period of time. Methods: We analyzed the dry weight, colony forming unit (CFU) number, extracellular polysaccharide (EPS) biovolume, and acid production rate of S. mutans biofilms formed on saliva-coated hydroxyapatite discs after 26 h, 50 h, 74 h, 98 h, 171 h, and 195 h. In addition, we performed a laser scanning confocal fluorescence microscopy to determine the bacterial volume, EPS biovolume, and biofilm thickness. We calculated the biofilm density using dry weight and EPS biovolume. Results: Over a period of time, there was no change in the CFU number and acid production rate of S. mutans biofilms, but there was an increase in the dry weight and EPS biovolume of S. mutans biofilms. The bacterial volume, EPS biovolume, and biofilm thickness only increased in the 50-h-old biofilm; however, no change was observed in 50-195-h-old biofilms. In addition, an increase in the biofilm density was observed over time. Conclusions: These results suggest that the acid production ability of cariogenic biofilms does not change, but the biofilm density increases over time. However, due to scientific information, further research needs to be conducted in the field of dentistry to get further insights on the progression of cariogenic biofilms over time.

• Journal of Microbiology and Biotechnology
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• v.27 no.4
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• pp.685-693
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• 2017

Candidiasis involving the biofilms of Candida albicans is a threat to immunocompromised patients. Candida biofilms are intrinsically resistant to the antifungal drugs and hence novel treatment strategies are desired. The study intended to evaluate the anti-Candida activity of allyl isothiocyanate (AITC) alone and with fluconazole (FLC), particularly against the biofilms. Results revealed the concentration-dependent activity of AITC against the planktonic growth and virulence factors of C. albicans. Significant (p <0.05) inhibition of the biofilms was evident at ${\leq}1mg/ml$ concentrations of AITC. Notably, a combination of 0.004 mg/ml of FLC and 0.125 mg/ml of AITC prevented the biofilm formation. Similarly, the preformed biofilms were significantly (p <0.05) inhibited by the AITC-FLC combination. The fractional inhibitory concentration indices ranging from 0.132 to 0.312 indicated the synergistic activity of AITC and FLC against the biofilm formation and the preformed biofilms. No hemolytic activity at the biofilm inhibitory concentrations of AITC and the AITC-FLC combination suggested the absence of cytotoxic effects. The recognizable synergy between AITC and FLC offers a potential therapeutic strategy against biofilm-associated Candida infections.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.473-486
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• 2019

In the aquatic environment, microorganisms are predominantly organized as biofilms. Biofilms are formed by the aggregation of microbial cells and are surrounded by a matrix of extracellular polymeric substances (EPS) secreted by the microbial cells. Biofilms are attached to various surfaces, such as the living tissues, indwelling medical devices, and piping of the industrial potable water system. Biofilms formed from a single species has been extensively studied. However, there is an increased research focus on multispecies biofilms in recent years. It is important to assess the microbial mechanisms underlying the regulation of multispecies biofilm formation to determine the drinking water microbial composition. These mechanisms contribute to the predominance of the best-adapted species in an aquatic environment. This review focuses on the interactions in the multispecies biofilms, such as coaggregation, co-metabolism, cross-species protection, jamming of quorum sensing, lateral gene transfer, synergism, and antagonism. Further, this review explores the dynamics and the factors favoring biofilm formation and pathogen transmission within the drinking water distribution systems. The understanding of the physiology and biodiversity of microbial species in the biofilm may aid in the development of novel biofilm control and drinking water disinfection processes.

• Environmental Engineering Research
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• v.19 no.3
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• pp.275-281
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• 2014

Aim of this study was to investigate biofilms attached in pipeline of water reuse from the MBR system treating sewage without chlorination in correlation to the outflow water quality. Two general pipe materials: polyvinyl chloride (PVC) and polyethylene (PE) were employed in the experiment. The peak growths were found at week 4 in both pipes. The maximum biofilms in PE pipe was $33mgVSS/cm^2$ with the growth rate of $4.75mgVSS/cm^2-d$ which was significant higher than that of PVC pipe. Biofilms examined by PCR-DGGE technique revealed five bacterial species in PE biofilms namely Sinorhizobium medicae WSM419, Sinorhizobium fredii NGR234, Geobacter sp. M18, Parachlamydia acanthamoebae UV-7, and Mycobacterium chubuense NBB4. The VSS concentrations in outflow had directly correlated to the biofilm attachment and detachment. High COD concentrations of outflow appeared during biofilm detaching phase. In summary, water quality of reuse water corresponded to the biofilms attachment and detachment in the pipeline.

• Journal of Periodontal and Implant Science
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• v.44 no.2
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• pp.79-84
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• 2014

Purpose: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. Methods: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. Results: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. Conclusions: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.