• 대한미생물학회지
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• 제35권1호
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• pp.77-85
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• 2000

Streptococcus mutans is the most important causative bacteria of dental caries among the oral bacteria. Lactococcus lactis 1370 was isolated from the oral cavity of child. The effect of Lactococcus lactis 1370 on the formation of artificial plaque by Streptococcus mutans was studied. 1. The insoluble substances and bacteria were much more attached on the wall of disposable cuvette in the culture of Streptococcus mutans than in the combined culture of Streptococcus mutans and Lactococcus lactis 1370. 2. The mean weight of produced artificial plaque on the wires in the beaker was 131.7 mg in the culture of Streptococcus mutans only, whereas being reduced to 6.4 mg in the combined culture of Streptococcus mutans and Lactococcus lactis 1370 (p<0.05). The viable cell didn't show the significant difference between them after culturing. 3. When Streptococcus mutans was cultured in the media containing culture supernatant of Lactococcus lactis 1370 cultured in M17 broth containing 0.5% yeast extract and 5% sucrose, the mean weight of produced artificial plaque was 8.0 mg on the wires, whereas being 125.4 mg in the media without culture supernatant of Lactococcus lactis 1370 (p<0.05). The viable cell didn't show the significant difference between them after culturing. 4. When Streptococcus mutans was cultured in the media containing soluble polymer produced by Lactococcus lactis 1370, the mean weight of produced artificial plaque was significantly reduced compared with being cultured in the media without soluble polymer (p<0.05). The viable cell didn't show the significant difference between them after culturing. 5. The soluble polymer produced by Lactococcus lactis 1370 was glucan. 6. The glucan produced by Lactococcus lactis 1370 was water-soluble glucan containing ${\alpha}$-1,6-glucose linkage as the main linkage. These results suggest that the artificial plaque formed by Streptococcus mutans is inhibited by water-soluble glucan produced by Lactococcus lactis 1370.

• 한국축산식품학회지
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• 제31권6호
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• pp.851-857
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• 2011

This study was conducted to develop a real time reverse transcriptase-PCR (RT-PCR) method for the detection of viable Enterobacteriaceae in milk using primers based on the genes of ribosomal proteins S11 and S13 and to determine effects of heating and subsequent treatments on the threshold cycle (Ct) of the real time RT-PCR. Total RNA was isolated from 17 strains of bacteria including 11 strains of Enterobacteriaceae suspended in milk using a modified Tri reagent method. SYBR Green Master Mix was added to the RNA and the mixture was subjected to the real time RT-PCR. The Cts of eleven type strains of the Enterobacteriaceae in milk ($10^7$ cells) in the real time RT-PCR ranged from 21.5 to 24.6. However, the Cts of Pseudomonas fluorescens, Acinetobacter calcoaceticus, and three gram-positive bacteria were more than 40. The real time RT-PCR detected as low as $10^3$ cells in agarose gel electrophoresis. The Cts increased from 22.0 to 34.2 when milk samples contaminated with Escherichia coli ($10^7$ cells/mL) were heated at $65^{\circ}C$ for 30 min. In addition, subsequent incubation at $37^{\circ}C$ for 6 and 24 h increased the Cts further up to 36.2 and 37.2, respectively. Addition of RNase A to the bacterial suspension obtained from the heated milk and subsequent incubation at $37^{\circ}C$ for 1 h increased the Cts to more than 40. The results of this study suggests that pretreatment of bacterial cells heated in milk with RNase A before RNA extraction might enhance the ability to differentiate between viable and dead bacteria using real time RT-PCR.

• Preventive Nutrition and Food Science
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• 제9권1호
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• pp.7-13
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• 2004

Lactic acid fermentation of prickly pear extract (PPE) was performed by Lactobacillus rhamnosus LS, Lactobacillus bulgaricus, and Lactobacillus brevis. The PPE was pasteurized to eliminate indigenous microorganisms as well as to dissolve the partially insoluble pulp. The PPE fermented without yeast extract by L. rhamnosus LS exhibited 0.57％ acidity and 3.5${\times}$10$^{8}$ CFU/mL bacteria count. With the addition of 0.2% edible yeast extract the PPE fermented by L. rhamnosus LS exhibited 1.15％ acidity,2.7${\times}$10$^{9}$ CFU/mL bacteria count and 95.0% retention of red color. When 5％ fructose syrup was added, the PPE fermented by L. rhamnosus LS had 1.09% acidity, 6.5${\times}$10$^{8}$ CFU/mL, and 97.7% retention of red color. With 1∼3% (w/v) concentrations of starter, the PPE fermented by L. bulgaricus and L. brevis showed 0.97% and 0.65% acidities, respectively. The viable cell counts from L. rhamnosus LS fermentation were higher compared with those of other LAB. During cold storage at 4$^{\circ}C$, the viable cell count was well maintained for 3 weeks, but then rapidly decreased. The red pigment was highly stable during cold storage for 4 weeks. The pasteurized PPE fortified with 5% fructose syrup, 0.2% yeast extract, and 0.05% CaCO$_3$ was successfully fermented by inoculating with 3% LAB and incubating at 3$0^{\circ}C$ for 2 days. Both viable cell counts and the red color of the fermented PPE were well maintained during cold storage for 3 weeks.

• 미생물학회지
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• 제42권2호
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• pp.116-124
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• 2006

직접 생균수 측정법(DVC)과 평판계수법(PC)을 이용하여 소나무림과 상수리나무림 토양에 분포하는 세균군집의 정량적 평가를 실시한 결과, DVC법에 의해 계수된 생균수에 대해 평판법에 의해 계수된 생균수 1% 미만으로 나타났다. 이상의 결과로부터 산림토양 내에 평판배양법으로는 배양이 곤란한 난배양성(viable but non culturable; VBNC) 세균이 99% 이상 존재해 있는 것으로 판단되었다. 이들 VBNC 세균의 군집구조 해석을 위하여 토양으로부터 직접 DNA를 추출하고 16S rDNA-ARDRA 분석을 통하여 계통학적 특성을 검토하였다. 소나무림과 삼수리나무림 토양으로부터 각각 111 clones, 108 clones을 획득하고 HaeIII 절편양상에 따라 30 groups과 26 groups의 ARDRA group으로 분류하였다. 각 ARDRA group으로부터 대표 clone을 선발하여 16S rDNA 염기서 열을 결정한 결과, 소나무림 토양의 경우 ${\alpha}$-proteobacteria (12 clones), ${\gamma}$-proteobacteria (3 clones), ${\delta}$-proteobncteria (1clone), Flexibacter/Cytophaga (1 clone), Actinobacteria (4 clones), Acidobacteria (4 clones), 그리고 Planctomycetes (5 clone)의 7개의 계통군이 확인되었으며, 상수리나무림 토양에서는 ${\alpha}$-proteobacteria (4 clones), ${\gamma}$-proteobacteria (2 clones), Actinobacteria (10 clones), Acidobacteria (8 clones), Planctomycetes (1 clone), 그리고 Verrucomicrobia (1clone)로 6개의 다양한 계통군이 확인되었다. 이상, 소나무림과 상수리나무림 토양 내에 존재하는 99% 이상의 VBNC 세균군집의 대부분은 미배양성 혹은 미동정균으로 계통학적으로 다양한 미지의 미생물로 구성되어 있음이 확인되었다.

• 한국식품저장유통학회지
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• 제16권6호
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• pp.965-970
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• 2009

유통중인 계란의병원성균 오염상태를 시판되는 multiplex polymerase chain reaction (mPCR) kit로 검사한 결과, 총 90개의 검사시료 가운데 한 가지 이상의 미생물이 검출된 계란시료는 30개로, 검사시료의 미생물 오염도는 약 33.3% 수준이었다. 미생물별 검출 빈도는 B. cereus가 17개 시료(18.9%)에서 검출되어 가장 높은 빈도를 보였으며 Y. enterocolitica는 16개(17.8%), L. monocytogenes 15개(16.7%), St. aureus 12개(13.3%), E. coli O157:H7 4개(4.4%)의 검출빈도를 보였다. 한편, 선택배지를 이용한 viable cell count 방법에서는 27개의 시료에서 미생물이 검출되어 검사시료의 미생물 오염도는 약 30.0% 수준이였으며, 미생물별 검출 빈도는 B. cereus가 17개 시료(18.9%)에서 14개(15.6%) 시료로, Y. enterocolitica는 16개(17.8%)에서 12개(13.3%)로, L. monocytogenes는 15개(16.7%)에서 13개(14.4%)로, St. aureus는 12개(13.3%)에서 10개(11.1%)로 감소하였으며, E. coli O157:H7은 두 가지 검출방법간의 차이가 나타나지 않았다. 검출 대상 미생물 9종 가운데 Campylobacter jejuni, Vibrio parahaemolyticus, Salmonella spp., 및 Shigella spp.는 두 방법 모두에서 검출되지 않았다. 이상의 결과에서 살펴본 바와 같이 PCR을 이용한 병원성 미생물의 검출 방법은 viable cell count 방법보다 감도가 높음을 알 수 있었다.

• Animal Bioscience
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• 제35권2호
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• pp.281-289
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• 2022

Objective: The aim of this study was to characterize the exopolysaccharides (EPS)-producing lactic acid bacteria from Taiwanese ropy fermented milk (TRFM) for developing a clean label low-fat fermented milk. Methods: Potential isolates from TRFM were selected based on the Gram staining test and observation of turbid suspension in the culture broth. Random amplified polymorphic DNA-polymerase chain reaction, 16S rRNA gene sequencing, and API CHL 50 test were used for strain identification. After evaluation of EPS concentration, target strains were introduced to low-fat milk fermentation for 24 h. Fermentation characters were checked: pH value, acidity, viable count, syneresis, and viscosity. Sensory evaluation of fermented products was carried out by 30 volunteers, while the storage test was performed for 21 days at 4℃. Results: Two EPS-producing strains (APL15 and APL16) were isolated from TRFM and identified as Lactococcus (Lc.) lactis subsp. cremoris. Their EPS concentrations in glucose and lactose media were higher than other published strains of Lc. lactis subsp. cremoris. Low-fat fermented milk separately prepared with APL15 and APL16 reached pH 4.3 and acidity 0.8% with a viable count of 9 log colony-forming units/mL. The physical properties of both products were superior to the control yogurt, showing significant improvements in syneresis and viscosity (p<0.05). Our low-fat products had appropriate sensory scores in appearance and texture according to sensory evaluation. Although decreasing viable cells of strains during the 21-day storage test, low-fat fermented milk made by APL15 exhibited stable physicochemical properties, including pH value, acidity, syneresis and sufficient viable cells throughout the storage period. Conclusion: This study demonstrated that Lc. lactis subsp. cremoris APL15 isolated from TRFM had good fermentation abilities to produce low-fat fermented milk. These data indicate that EPS-producing lactic acid bacteria have great potential to act as natural food stabilizers for low-fat fermented milk.

• Preventive Nutrition and Food Science
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• 제9권3호
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• pp.206-212
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• 2004

Prickly pear extract (PPE) was fermented by Lactobacillus rhamnosus LS at 3$0^{\circ}C$ for 2 days. To improve the physicochemical properties of fermented PPE, it was fortified with food polysaccharides (0.2 %) or seaweed calcium before lactic acid fermentation. The viable cell counts, flow behavior, titratable acidity and color stability of fermented PPE were evaluated during 4 weeks of cold storage. Addition of xanthan gum or glucomannan increased the apparent viscosity and acid production, viable cell counts and red color of PPE were also well maintained during the cold storage. However, fermenting PPE with gellan gum resulted in a decrease in relative absorbance, indicating lower color stability. In particular, PPE fortified with carrageenan or alginic acid showed reduced acid production and lower viable cell counts. Addition of seaweed calcium at a 0.1 % level had positive effects on color stability, and helped maintain viable cell counts of 4.1 ${\times}$ 10$^{9}$ CFU/mL. This study demonstrated that xanthan gum could be used as a good thickening agent and stabilizer for retaining viable cell counts and red color during the cold storage in PPE fermented by lactic acid bacteria.

• 한국수산과학회지
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• 제55권4호
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• pp.431-436
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• 2022

For the safety assessment of microbiological and chemical hazards in katsuobushi, fifteen samples of katsuobushi were purchased from supermarkets. The contamination levels of total viable bacteria, coliforms, Escherichia coli, and nine pathogenic bacteria [Staphylococcus aureus, Salmonella spp., Listeria monocytogenes, Bacillus cereus, Vibrio parahaemolyticus, Clostridium perfringens, Enterohemorrhagic E. coli (EHEC), Yersinia enterocolitica and Campylobacter jejuni/coli] were quantitatively or qualitatively assessed. Additionally, the heavy metals (total and methyl mercury) content, radioactivity (¹³¹ I, ¹³⁴ Cs⁺ and ¹³⁷ Cs) were quantitatively assessed. Microbial and chemical analyses were performed using standard methods in Korean food code. The contamination level of total viable bacteria was 2.70 (1.18-4.42) log CFU/g. Coliforms, E. coli and S. aureus were not detected in any samples. Other eight pathogenic bacteria were negative in all samples. The contamination levels of total and methyl mercury were 0.366 (0.227-0.481) and 0.120 (0.002-0.241) mg/kg, respectively. In addition, radioactivity was not detected in any samples. The results will be helpful in revitalizing domestic use and boosting exports of katsuobushi because the microbiological and chemical safety of katsuobushi has been assured. Furthermore, the results may be used as a basis for performing chemical and microbial risk assessments of katsuobushi.

• Journal of Animal Science and Technology
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• 제46권5호
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• pp.841-848
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• 2004

살균된 우유에 오염된 Salmonella의 신속한 검출 방법을 공중 보건의 보호를 위하여 중요하다. RT-PCR 방법은 생균과 사균을 분별하여 검출할 수 있는 분자유전학적 방법이다. 본 연구의 RT-PCR 방법은 Salmonella의 type 1 fImbriae의 단량체 단백질을 암호화하는 fimA 유전자의 mRNA를 주형으로 하여 DNA를 증폭하는 방법으로서 활력이 있는 Salmonella를 검출할수 있기 위하여 개발되었다. RNA 시료를 RQRNase-free DNase로 처리하여 오염된 DNA를 파괴하여 RT-PCR 반응에서 주형으로서 DNA가 합성되는 것을 방지할 수 있었다. 이 RT-PCR 방법은 Salmonella 7균주를 검출하였으나 Escherichia coli, Shigella sonnei, Citrobacter freundii, 및 Klebsiella pneumoniae 는 검출하지 않았다. 우유 내에서 열처리된 $10^7/ml과 10^6/ml$ 세균수의 Salmonella는 검출되 었으나 $10^5{\sim}100/ml$는 검출되지 않았다. RT-PCR를 검출할 수 있는 활력이 있는 Salmonella의 최소 세균수는 100/ml이었다.

• Food Science and Biotechnology
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• 제14권4호
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• pp.547-550
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• 2005

Fourteen out of 87 strains of lactic acid bacteria isolated tram Kimchi were found to be resistant against the action of artificial gastric and bile juices. In particular, lactobacilli KM 3, 7, 28, and 37 showed strong resistance and their viable cell counts at the initial stage remained the same even after 3 hours of cultivation in an artificial gastric juice. However, the survival rates of KM 14, 28, and 64 were found to be significantly enhanced in artificial bile juice. Based on the paper disc method, it was evident that isolated lactic acid bacteria showed antibacterial effect against Listeria monocytogenes, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Vibrio vulnificus, and Salmonella typhimurium. The isolated lactic acid bacteria were identified as Lactobacillus plantarum and Leuconostoc mesenteroides.