• Microbiology and Biotechnology Letters
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• v.35 no.1
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• pp.17-25
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• 2007

The kinds and quantity of antimicrobial agents used for cattle (animal industry) may be considerable, suggesting the possibility that pathogenic bacteria which cannot be extirpated by the existing antimicrobial agents could appear. Ten cattle, pig and chicken farms, respectively, were randomly selected from 5 provinces in Korea and the samples were collected from excrement, manure, underground water, farmers' hands and the neishboring environment. h total of 299 samples were examined and 197 of Escherichia coli, 13 of Campylobacter jejun/coli, 223 of Enterococcus faecium/faecalis and 42 of Staphylococcus aureus isolates were collected. All isolates were screened for antimicrobial resistance: 69.4% of E. coli (137/197 strains), 78.6% of S. aureus (33/42 strains), and 82.1% of E. faecium/faecalis (183/223 strains) were resistant to one antimicrobial agent and all of C. jejuni/coli Isolates showed the resistance to one antimicrobial agent. Meanwhile, the multiple resistance ratio for more than 4 lines of antimicrobial agent was 19.2% of E. coli (38/197 strains), 11.9% of S. aureus (5/42 strains), 15.4% of C. jejuni/coli (2/13 strains) and 6.2% of E. faecium/faecalis (14/223 strains). The antimicrobial resistance ratio of bacteria isolated from the cattle farm showed lower than that of bacteria isolated from the pig or chicken farm, which might be related to the quantify of antimicrobial agents consumed. And one strain of vancomycin resistant E..faecium (VREF) were isolated from the excrement of chicken and stream, respectively. Generally, the ratio of VREF collected in animal farm environments is lower than that of VREF collected in medical environment.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.250-256
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• 2017

The prevalence of infections due to pathogenic bacteria such as Edwardsiella tarda, Streptococcus parauberis, and Photobacterium phosphoreum in fish farms in Jeju Island and their management by marine-derived biomaterials was studied. In this study, we isolated eight spices type of marine-derived biomaterials from four sea areas of Jeju Island. An antibiotic disc susceptibility test confirmed that the isolated marine-derived biomaterials showed weak resistance only to oxytetracycline and penicillin and sensitivity to the other antibiotics tested, and antimicrobial activity against fish pathogens with the inhibitory zone of 22 mm, 18 mm, and 19 mm for MD-02, MD-04, and MD-06 against E. tarda strains, respectively, and 19 mm, 22 mm, 30 mm, and 29 mm for MD-01, MD-02, MD-04, and MD-06 against S. parauberis strains, respectively, while all the marine-derived biomaterials showed antibacterial activity against P. phosphoreum. Among the eight biomaterials selected, Bacillus subtilis MD-02 displayed the greatest antibacterial activity against the three tested fish pathogens and also displayed susceptibility to antibiotics. The growth of Bacillus subtilis MD-02 was greatest with the carbon source, dextrine; nitrogen source, peptone; and mineral source, $MgSO_4{\cdot}7H_2O$. Hence, the present study confirmed that the isolate B. subtilis MD-02 from Jeju Island could be a potential antimicrobial agent against fish pathogens and a potential pharmacotherapeutic agent.

• Journal of Life Science
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• v.24 no.4
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• pp.361-369
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• 2014

Broad-range and specific 16S rRNA gene PCR is used for detection and identification of bacterial pathogens in clinical specimens from patients with a high suspicion for infection. We describe the development of a broad-range and specific PCR primer, based on bacterial 16S rRNA, for use in routine diagnostic clinical microbiology services. The primers were designed by using conservative regions of 16S rRNA sequences from 10 strains. Ninety-eight clinical strains were isolated from clinical patient specimens. A total of 98 strains of bacteria were identified by phenotypic methods; PCR with newly designed primers and universal primers. All purified PCR products were sequenced using both forward and reverse primers on an automated DNA analyzer. In this study, we evaluated the usefulness of the newly designed primers and the universal primers for the detection of bacteria, and both these techniques were compared with phenotypic methods for bacteria detection. When we also tested 98 strains of clinical isolates with newly designed primers, about 778 bp DNA fragments were amplified and identified from all strains. Of the 98 strains, 94 strains (95.9%) correspond in comparison with phenotypic methods. The newly designed primers showed that the identities of 98 (100%) strains were the same as those obtained by universal PCR primers. The overall agreement between the newly designed primers and universal primers was 100%. The primer set was designed for rapid, accurate, and cheap identification of bacterial pathogens. We think the newly designed primer set is useful for the identification of pathogenic bacteria.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.503-513
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• 1986

Previous studies have developed the technique of topical application of tetracycline(TC) into the periodontal pockets and examined the change of clinical parameters and subgingival microbial morphotypes. The purpose of this study was to longitudinally examine the clinical and microbiological effects of topically applied TC in a double-blind and split-mouth design. Thirteen patients with moderate periodontitis, who were treated with or without TC application and scaling treatment, were examined. TC gel(3%) was used to apply into the selected periodontal pockets twice a week for 2 weeks. During the experiment, clinical parameters and subgingival microbial morphotypes were examined, and for isolation of black-pigmented Bacteroides(BPB) and streptococci, an anaerobic sample culturing was done at week 0, 2, and 7. In clinical observation the TC-scaled group exhibited a significant decrease of Gingival Inflammatory Index, Plaque Index, Sulcus Bleeding Index, pocket depth, and gingival crevicular fluid when compared to the TC-unsealed, placebo-scaled, and placebo-unsealed groups. The result of microbial morphotype observation showed a significant increase of coccal form and a decrease of spirochetes in the TC-scaled, TC-unscaled, and placebo-scaled groups. The culture study of streptococci revealed that TC with scaling treatment resulted in a significant increase of S. sanguis I at week 2, but its proportion had returned to the base line level. The anaerobic culture study showed that BPB was significantly reduced in the TC-scaled and TC-unsealed groups at week 7. Among BPB species, B. intermedius declined significantly with time treatment(week 2 and 7) in the TC-scaled and TC-unsealed groups. These results suggest that the settled pathogenic microflora can be succeeded by nonpathogenic microflora in periodontal pockets after TC treatment.

• Microbiology and Biotechnology Letters
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• v.35 no.3
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• pp.196-202
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• 2007

Mutations in the cystathionine ${\beta}$-synthase (CBS) gene cause homocystinuria, the most frequent inherited disorder in sulfur metabolism. CBS is the unique enzyme using both heme and pyridoxal 5-phosphate (PLP) for activity. Among the reported 140 mutations, one of the most common disease-causing alterations in human CBS is G307S mutation. To investigate the pathogenic mechanism of G307S by spectroscopic methods, we engineered the full length and the truncated G247S mutation of yeast CBS that is corresponding mutation to human G307S. Yeast CBS does not contain heme and thus gives a merit to study the spectroscopic properties. The UV-visible spectra of the purified full length and the truncated G247S yeast CBSs showed the total absence of PLP in the protein. The absence of PLP in G247S mutation was also confirmed by the PLP-cyanide adduct formation experiment, which was conducted by the incubation of the purified enzyme with KCN. The adducts were detected using a circular dichroism (CD) and a spectrofluorimeter. Radio isotope activity assay of full length and truncated G247S proteins also gave no activity. Our yeast G247S mutation data suggested that G307S might make the distortion of the active site so that cofactor PLP and substrate can not fit inside the active site. Our yeast CBS study addressed the reason why the G307S mutation in human CBS makes the enzyme inactive that consequently leads to severe clinical phenotype.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.190-197
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• 2010

To develop the safe and natural antimicrobial agents, the 68 ethanol extracts from the 61 different kinds of oriental herbal medicine were prepared and their antimicrobial activities were evaluated. The herbal medicine used were from China (46 kinds), South Korea (14 kinds), North Korea (5 kinds) and Vietnam (3 kinds), respectively, and the root (27 species) was popular part in this study. The average water content and extraction ratio for ethanol were 7.10% and 6.75%, respectively. Determination of antimicrobial activity by disc-diffusion assay at 0.5 mg/disc concentration showed that the extract of Angelica tenuissima Nakai (china), Illicium verum, Junci medulla, Rhus javanica L., Salvia miltiorrhiza Bunge and Syzygium aromaticum has strong antimicrobial activities against different food spoilage and pathogenic bacteria and fungi. Determination of MIC and MBC/MFC further showed that the extract of Syzygium aromaticum has MIC of 1.25 mg/mL and MBC/MFC of 1.25~5.00 mg/mL against Listeria monocytogenes, Bacillus subtilis, Staphylococcus epidermidis, Staphylococcus aureus, Salmonella typhimurium, Proteus vulgaris, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Saccharomyces cerevisiae. And, the extract of Junci medulla, Rhus javanica L. and Salvia miltiorrhiza Bunge showed strong antibacterial activities with MIC of 0.08~0.63 mg/mL and MBC/MFC of 0.08~2.50 mg/mL against the tested bacteria except E. coli and P. aeruginosa. In a while, the results of hemolytic activity of 68 different herbal extracts against human red blood cells showed that the extract of Angelica tenuissima Nakai has hemolytic activity at 0.5 mg/mL concentration. Therefore, Illicium verum, Junci medulla, Rhus javanica L., Salvia miltiorrhiza Bunge and Syzygium aromaticum were finally selected for natural antimicrobial resources. Further research on active substances and the mode of action of the selected herbal medicine is necessary.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.172-178
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• 2013

The strains were added to the feed in the concentration of $10^3$, $10^5$, and $10^7$ CFU/kg and 2% of fishes were given the feed twice a day (8 AM and 5 PM) for 12 weeks. In result of the nonspecific immune response study to examine Respiratory burst activity, Lysozyme activity and Phagocytosis activity every two weeks until the end of the study, all test samples showed greater activities than control samples and improved immune activity with Bacillus sp. IS-2. The mortality test performed by artificial infection using Streptococcus iniae, a pathogenic bacterium, after the completion of this study also showed over 55% greater survival rate in all test samples. In result of performing PCR using the universal primer to verify that the probiotic stays in the intestines of the fishes, all test samples showed PCR product of 1,465 bp. Based on the above findings, it was concluded that Bacillus sp. IS-2 in the feed improved farmed flatfish's immune system and resistance against diseases as the probiotics. Also, the physiological indicators discovered by this study would be useful for identifying the mechanisms of probiotics.

• Korean Journal of Microbiology
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• v.46 no.4
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• pp.334-340
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• 2010

It is well known that smoking as well as drinking is a factor of stomatopathy, however there are few investigations about comparison of oral flora between smokers and non-smokers. In this study, we isolated the oral flora of 30 smokers and 30 non-smokers and cultured them on blood agar plates. The isolated pathogenic microorganisms were tested for antibiotic susceptibility and resistance using the Kirby-Bauer antibiotic testing method. Each colony was stained using the Gram staining method and was identified by an automatic identifier, known as the VITEK system. We isolated 41 colonies from smokers' oral cavity, and they were sorted as 63% of Gram-positive cocci, 29% of Gram-negative cocci, 3% of Gram-positive bacilli, and 5% of Gram-negative bacilli by gram staining, whereas 38 colonies were isolated from non-smoters' oral cavity, and their proportions were 55% of Gram-positive cocci, 26% of Gram-negative cocci, 3% of Gram-positive bacilli, and 16% of Gram-negative bacilli. The VITEK system revealed specific distribution of bacteria species that Streptococcus mutans (6/41), Gemella morillorum (6/41), Streptococcus oralis (2/41), Streptococcus pneumoniae (1/41), Staphylococcus aureus (3/41), Streptococcus anginosus (1/41), Streptococcus intermedius (1/41), Streptococcus uberis (1/41), and Streptococcus sanguinis (1/41) in smokers oral cavity whereas Streptococcus sanguinis (8/38), Staphylococcus aureus (1/38), Staphylococcus auricularis (1/38), Streptococcus uberis (1/38), Streptococcus intermedius (1/38), Streptococcus mutans (1/38), and Streptococcus oralis (1/38) in those of non-smokers'. Three cases of Staphylococcus aureus from smokers produced Beta-lactamase and were identified methicillin-resistance Staphylococcus aureus (MRSA). However one case of Staphylococcus aureus from non-smoker did not produce Beta-lactamase and was sensitive to methicillin. In conclusion, the distribution of oral flora was different between smokers' and non-smokers' oral cavity, especially Gemella morillorum and MRSA were predominantly found in smoker's oral cavity. These results are useful in the treatment and prevention of patients with stomatopathy caused by smoking.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.140-147
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• 2016

A process for manufacturing virally-safe porcine bone hydroxyapatite (HA) has been developed to serve as advanced xenograft material for dental applications. Porcine bone pieces were defatted with successive treatments of 30% hydrogen peroxide and 80% ethyl alcohol. The defatted porcine bone pieces were heat-treated in an oxygen atmosphere box furnace at $1,300^{\circ}C$ to remove collagen and organic compounds. The bone pieces were ground with a grinder and then the bone powder was sterilized by gamma irradiation. Morphological characteristics such as SEM (Scanning Electron Microscopy) and TEM (Transmission Electron Microscopy) images of the resulting porcine bone HA (THE Graft$^{(R)}$) were similar to those of a commercial bovine bone HA (Bio-Oss$^{(R)}$). In order to evaluate the efficacy of $1,300^{\circ}C$ heat treatment and gamma irradiation at a dose of 25 kGy for the inactivation of porcine viruses during the manufacture of porcine bone HA, a variety of experimental porcine viruses including transmissible gastroenteritis virus (TGEV), pseudorabies virus (PRV), porcine rotavirus (PRoV), and porcine parvovirus (PPV) were chosen. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the $1,300^{\circ}C$ heat treatment. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.81$$ for PRV, $${\geq_-}6.28$$ for PRoV, and $${\geq_-}5.21$$ for PPV. Gamma irradiation was also very effective at inactivating the viruses. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the gamma irradiation. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.87$$ for PRV, $${\geq_-}6.05$$ for PRoV, and $${\geq_-}4.89$$ for PPV. The cumulative log reduction factors achieved using the two different virus inactivation processes were $${\geq_-}9.30$$ for TGEV, $${\geq_-}11.68$$ for PRV, $${\geq_-}12.33$$ for PRoV, and $${\geq_-}10.10$$ for PPV. These results indicate that the manufacturing process for porcine bone HA from porcine-bone material has sufficient virus-reducing capacity to achieve a high margin of virus safety.

• Journal of Food Hygiene and Safety
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• v.38 no.3
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• pp.123-130
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• 2023

Salmonella spp. are prevalent foodborne pathogens that are infective at relatively low concentrations, thus posing a serious health threat, especially to young children and the elderly. In several countries, the management and regulation of Salmonella spp. in food, including seafood, adhere to a negative detection standard. The risk of infection is particularly high when seafood is consumed raw, which underscores the importance of timely detection of pathogenic microorganisms, such as Salmonella. Accordingly, this study aimed to develop a combined pre-treatment and detection method that includes pre-culture and DNA extraction in order to detect five species of Salmonella at concentrations below 10 CFU/mL in seafood. The effectiveness of the proposed method was assessed in terms of the composition of the enrichment (pre-culture) medium, minimum incubation time, and minimum cell concentration for pathogen detection. Furthermore, a practical DNA extraction method capable of effectively handling high salt conditions was tested and found to be successful. Through polymerase chain reaction, Salmonella spp. Were detected and positively identified in shellfish samples at cell concentrations below 10 CFU/g. Thus, the proposed method, combining sample pre-treatment and cell culture with DNA extraction, was shown to be an effective strategy for detecting low cellular concentrations of harmful bacteria. The proposed methodology is suitable as an economical and practical in situ pre-treatment for effective detection of Salmonella spp. in seafood.