• Journal of Microbiology and Biotechnology
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• v.33 no.8
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• pp.1101-1110
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• 2023

Streptococcus mutans is the primary causative agent of caries, which is one of the most common human diseases. Thus, rapid and early detection of cariogenic bacteria is critical for its prevention. This study investigated the combination of loop-mediated isothermal amplification (LAMP) and microfluid technology to quantitatively detect S. mutans. A low-cost, rapid microfluidic chip using LAMP technology was developed to amplify and detect bacteria at 2.2-2.2 × 10⁶ colony-forming units (CFU)/ml and its detection limits were compared to those of standard polymerase chain reaction. A visualization system was established to quantitatively determine the experimental results, and a functional relationship between the bacterial concentration and quantitative results was established. The detection limit of S. mutans using this microfluidic chip was 2.2 CFU/ml, which was lower than that of the standard approach. After quantification, the experimental results showed a good linear relationship with the concentration of S. mutans, thereby confirming the effectiveness and accuracy of the custom-made integrated LAMP microfluidic system for the detection of S. mutans. The microfluidic system described herein may represent a promising simple detection method for the specific and rapid testing of individuals at risk of caries.

• Journal of the korean academy of Pediatric Dentistry
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• v.36 no.2
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• pp.237-244
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• 2009

When the symptom of periapical infection is not released by mechanical instrumentation. anti-microbial agents including antibiosis become necessary in order to remove microorganisms from the root canal. Since anti-microbial agents of natural origins are currently popular, more natural remedies are being sought out. As it turns out, it is well known isothiocyanates (ITCs) in horseradish root extract have anti-microbial activity from many studies. In this research, anti-microbial effects of horseradish root extract and chlorhexidine, a typical anti-microbial agent, were investigated and compared against two kinds of obligate anaerobes. Fusobacterium nucleatum and Prevotella nigrescens, that are often discovered in infected root canal, and Clostridium perfringens, which is resistant to antibiotics and frequently used as a control strain for antibacterial studies 1. The MIC and MBC of horseradish root extract were ranged from 87 to 470 ppm and from 156 to 625 ppm against three kinds of obligate anaerobes, respectively. Horseradish root extract showed the strongest anti-bacterial activity (MBC, 156 ppm) against F. nucleatum and also showed anti-bacterial activity against antibiotic resistant obligate anaerobes. C. perfringens. 2. The MIC and MBC of chlorhexidine were ranged from 3.12 to 6.25 ppm and 10.94 ppm against three kinds of obligate anaerobes, respectively. 3. The MIC with 87-470 ppm of horseradish root exact has the same growth inhibiting effect as the one of 3.12-6.25 ppm of chlorhexidine. Likewise, the MBC with 156-625 ppm of horseradish has the similar bactericidal effect as 10.94 ppm of chlorhexidine.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.22 no.2
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• pp.123-132
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• 2000

$TGF-{\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to infection control. The objective of this study is to investigate production of $TGF-{\beta}$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of $TGF-{\beta}_1$ which may be responsible for infection control. The fibroblasts were originated from gingiva and facial dermis in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.l{\mu}g$, $1.0{\mu}g$) respectively, $cells(5{\times}10^3ml)$ were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, $cells(2.5{\times}10^5ml)$ were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and $LPS(0.1{\mu}g)$ and $SEB(0.1{\mu}g)$ in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and $TGF-{\beta}_1$ was assayed in duplicate. The results were as follows. 1. In gingival fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell Proliferation occurred very significantly since 3 days after incubation, compared with the control and the production of $TGF-{\beta}_1$ occurred very significantly at 1 day after incubation, compared with the control. 2. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of $TGF-{\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of $TGF-{\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of $TGF-{\beta}_1$ very significantly. The gingival and facial dermal fibroblasts have different phenotype each other The orchestrated understanding of fibroblast proliferation and $TGF-{\beta}_1$ production play an important part in host defense against the bacterial Infection and may prevent tissue necrosis such as necrotizing fasciitis and life-threatening syndrome such as multiple organ failure.

• Journal of dental hygiene science
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• v.17 no.4
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• pp.283-289
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• 2017

The water discharged from dental unit waterlines (DUWLs) is heavily contaminated with bacteria. The development of efficient disinfectants is required to maintain good quality DUWL water. The purpose of this study was to establish a DUWL biofilm model using well-plates to confirm the effectiveness of disinfectants in the laboratory. Bacteria were obtained from the water discharged from DUWLs and incubated in R2A liquid medium for 10 days. The bacterial solution cultured for 10 days was made into stock and these stocks were incubated in R2A broth and batch mode for 5 days. Batch-cultured bacterial culture solution and polyurethane tubing sections were incubated in 12-well plates for 4 days. Biofilm accumulation was confirmed through plating on R2A solid medium. In addition, the thickness of the biofilm and the shape and distribution of the constituent bacteria were confirmed using confocal laser microscopy and scanning electron microscopy. The average accumulation of the cultured biofilm over 4 days amounted to $1.15{\times}10^7CFU/cm^2$. The biofilm was widely distributed on the inner surface of the polyurethane tubing and consisted of cocci, short-length rods and medium-length rods. The biofilm thickness ranged from $2{\mu}m$ to $7{\mu}m$. The DUWL biofilm model produced in this study can be used to develop disinfectants and study DUWL biofilm-forming bacteria.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.185-194
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• 2006

The aim of this study was to identify the bacteria isolated from endodontic lesions by cell culture and to determine the antimicrobial susceptibility of them against 8 antibiotics. The necrotic pulpal tissues were collected from 27 infected root canals, which were diagnosed as endodontic infection. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing $500{\mu}l\;of\;1{\times}PBS$. The sample solution was briefly mixed and plated onto a BHI-agar plate containing 5% sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 2 to 5 days. The bacteria grown on the agar plates were identified by comparison of 16S rRNA gene (rDNA) sequencing method at the species level. To test the sensitivity of the bacteria isolated from the infected root canals against 8 antibiotics, minimum inhibitory concentrations (MIC) were determined using broth dilution assay. The data showed that 101 bacterial strains were isolated and were identified. Streptococcus spp. (29.7%) and Actinomyces spp. (21.8%) were predominantly isolated. The 9 strains were excluded in antimicrobial susceptibility test because they were lost during the experiment or were not grown in broth culture. The percentage of bacteria susceptible for each antibiotic in this study was clindamycin, 87.0% (80 of 92); tetracycline, 75.0% (69 of 92); cefuroxime axetil, 75.0% (69 of 92); amoxicillin + clavulanic acid (5:1), 71.7% (66 of 92); penicillin G, 66.3% (61 of 92); erythromycin, 66.3% (61 of 92); amoxicillin, 44.6% (41 of 92); and ciprofloxacin, 31.5% (29 of 92). The susceptibility pattern of 8 antibiotics was dependent on the host of the bacteria strains rather than the kinds of bacterial species. These results indicate that antibiotic susceptibility test should be performed when antibiotics are needed for the treatment of infected root canals.

• Environmental Analysis Health and Toxicology
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• v.18 no.2
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• pp.121-129
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• 2003

The genotoxicity of pendimethalin [N-(l-ethylpropyl)-2, 6-dinitro-3, 4-xylidine, C$\_$13/H$\_$19/N$_3$O$_4$, M.W.=281.3, CAS No. 40487-42-1], one of selective herbicide, was evaluated in bacterial gene mutation system, chromosome aberration in mammalian cell system and in vivo micronucleus assay with rodent. In bacterial gene mutation assay, pendimethalin revealed dose-dependent mutagenic potential in 313 ∼ 5,000 ${\mu}$g/plate of Salmonella typhimurium TA 98 and TA 1537 both in the absence and presence of S-9 metabolic activation system, and TA 100 only in the absence of S-9 mixture. In the TA 1535, slight increase of revertant was also observed in the presence of S-9 metabolic activation system. No mutagenic potential was observed in the TA 1535 without metabolic activation system and TA l00 in the presence of S-9 mixture. In mammalian cell system using Chinese hamster lung (CHL) fibroblast, no clastogenicity of pendimethalin was observed both in the absence and presence of S-9 metabolic activation system in the concentration range of 2.32∼9.28 ${\mu}$g/ml. And also, in vivo bone marrow micronucleus assay, pendimethalin revealed no clastogenic potential in the dose range of 203∼810 mg/kg body weight after oral administration in mice. Consequently, in vitro chromosome aberration with mammalian cells and in vivo bone marrow micronucleus assay revealed no clastogenic potential of pendimethalin. However, pendimethalin revealed mutagenic potential in bacterial gene mutation assay.

• Journal of Periodontal and Implant Science
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• v.28 no.4
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• pp.691-703
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• 1998

The 16S rRNA analyzing method is a bacterial identification method that is useful in identifying bacteria which is difficult to do by other means. The following 7 types of bacteria which are Treponema, A. actinomycetemcomitans, P. gingivalis, Fusobacterium, B. forsythus, P. intermedia, P. micros were evaluated in order to study their distribution among patients with adult periodontitis. The 16S rRNA analyzing method was used to compare bacterial distribution among 3 groups. Subgingival plaque acquired from the affected sites(pocket depth ${\geq}6mm$) of 29 patients with adult periodontitis were grouped as the experimental group while plaque from the non-affected sites(pocket depth ${\leq}3mm$) were grouped as control 2 and finally plaque acquired from students with healthy periodontal tissues were grouped as control 1. The results are as follows ; 1. The distribution of Treponema was 12.5% for control 1, 21.4% for control 2 and 75.4% for the experimental group. For A. actinomycetemcomitans the distribution was 0.5%, 19.0%, 44.4% in respect to the order of groups mentioned above. P.gingivalis showed 10.5%, 43.1%, 94.0% distribution, Fusobacterium 33.0%, 48.3%, 81.0% distribution, B. forsythus 9.5%, 17.2%, 65.9% distribution, P. intermedia 1.0%, 12.1%, 26.3% distribution and finally P. micros 5.0%, 19.0%, 48.7% respectively. In all 7 types of bacteria, the experimental group showed higher bacterial distribution compared to the other two groups with statistically significant difference. 2. In the case of Treponema, A. actinomycetemcomitans, gingivalis,Fusobacterium, B. forsythus, P. intermedia, P. micros showed significant difference between control 1 and 2. These results suggest that the 16S rRNA analyzing method which was applied on Koreans for the first time could be utilized and useful in finding potential pathogens of periodontal disease.

• The Korean Journal of Food And Nutrition
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• v.26 no.3
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• pp.383-390
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• 2013

Mistlero C was shown to be non-genotoxic in a series of genotoxicity tests, including a bacterial reverse mutation test and a combined in vivo mammalian erythrocyte micronucleus test. In a bacterial reverse mutation assay, no significant increases in the number of revertant colonies, compared to the negative control, was detected in $5,000{\mu}g/plate$ of Mistlero C. In addition, with Mistlero C, no changes were shown in the number of micronucleated polychromatic erythrocytes (MNPCE) among 2,000 polychromatic erythrocytes compared to the negative control. Mistlero C was administered orally in rats to investigate acute toxicity. The $LD_{50}$ values in rats were above 2,000 mg/kg. In a repeated dose, 13-week, oral toxicity study conducted in rats, no compound-related adverse effects were shown at doses of Mistlero C of up to 1,000 mg/kg body weight/day. The results of these studies support the safe use of Mistlero C in food for human consumption.

• Animal Bioscience
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• v.34 no.9
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• pp.1466-1478
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• 2021

Objective: Ruminants are completely dependent on their microbiota for rumen fermentation, feed digestion, and consequently, their metabolism for productivity. This study aimed to evaluate the rumen bacteria of lactating yaks with different milk protein yields, using high-throughput sequencing technology, in order to understand the influence of these bacteria on milk production. Methods: Yaks with similar high milk protein yield (high milk yield and high milk protein content, HH; n = 12) and low milk protein yield (low milk yield and low milk protein content, LL; n = 12) were randomly selected from 57 mid-lactation yaks. Ruminal contents were collected using an oral stomach tube from the 24 yaks selected. High-throughput sequencing of bacterial 16S rRNA gene was used. Results: Ruminal ammonia N, total volatile fatty acids, acetate, propionate, and isobutyrate concentrations were found to be higher in HH than LL yaks. Community richness (Chao 1 index) and diversity indices (Shannon index) of rumen microbiota were higher in LL than HH yaks. Relative abundances of the Bacteroidetes and Tenericutes phyla in the rumen fluid were significantly increased in HH than LL yaks, but significantly decreased for Firmicutes. Relative abundances of the Succiniclasticum, Butyrivibrio 2, Prevotella 1, and Prevotellaceae UCG-001 genera in the rumen fluid of HH yaks was significantly increased, but significantly decreased for Christensenellaceae R-7 group and Coprococcus 1. Principal coordinates analysis on unweighted UniFrac distances revealed that the bacterial community structure of rumen differed between yaks with high and low milk protein yields. Furthermore, rumen microbiota were functionally enriched in relation to transporters, ABC transporters, ribosome, and urine metabolism, and also significantly altered in HH and LL yaks. Conclusion: We observed significant differences in the composition, diversity, fermentation product concentrations, and function of ruminal microorganisms between yaks with high and low milk protein yields, suggesting the potential influence of rumen microbiota on milk protein yield in yaks. A deeper understanding of this process may allow future modulation of the rumen microbiome for improved agricultural yield through bacterial community design.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.66.1-66
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• 2003

The aim of this study was to characterize the interactions of rice and Burkholderia glumae, a causal agent of bacterial grain rot of rice, at molecular levels using whole genomic sequences and to identify genes important for pathogenicity and symptom development. To do these, we sequenced whole genome of the bacterium and constructed cosmid clone profiles. We generated pools of mutants using various transposons and determined mutation sites by sequencing rescued plasmids. We focused on studying toxoflavin biosynthetic genes, quorum sensing regulation, and Hrp type III protein secretion systems. We found that two possible operons consisting of five genes are involved in toxoflavin biosynthesis and their expression is regulated by quorum sensing and LysR-type regulator, ToxR. We have isolated the nn PAI of B. glumae and characterized by mutational analyses. The hrp cluster resembled most the putative Type III secretion systems of B. pseudomallei, which is the causative agent of melioidosis, a serious disease of man and animals. The Hrp PAI core region showed high similarity to that of Ralstonia solanacearum and Xanthomonas campestris, however some aspects were dissimilar.