• Title/Summary/Keyword: 16S-rRNA

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A Microbiological Analysis and Hazard Factor Evaluation of Food Utensils and Fixtures of Food Service Operations in Schools (학교 집단급식소 내 식기류 및 집기류의 미생물학적 분석 및 위해요인 평가)

  • Park, Sung-Jun;Hong, Sung-Ho;Lee, Anne Ha-Young;Kim, Cheol-Ju;Kim, Su-Jin;Kim, Sung-Kyoon;Ko, Gwang-Pyo
    • Journal of Environmental Health Sciences
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    • v.37 no.5
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    • pp.376-386
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    • 2011
  • Objectives: The aim of this study was to evaluate the microbial hazards posed by food utensils and fixtures in food service operations at selected middle and high schools located in Seoul, Korea. Methods: We collected 200 samples of utensils and fixtures including cups, spoons/chopsticks, food trays and tables from five different schools in Seoul. Target microorganisms of this study were divided into two groups: total bacterial count and total coliform as indicators of microbial contamination and Bacillus cereus and Staphylococcus aureus as pathogens of food poisoning. We used selective media to quantify microbial concentration and 16S rRNA PCR assay for qualitative analysis. In addition, intensive interviews with nutritionists were conducted and observations were made to identify factors that may affect microbial contamination. Logistic regression analysis was employed to examine the relationship between the microbial concentration and operation characteristics of each operation. Results: The level of microbial concentration in school B and C were significantly lower than in school A, D and E (p<0.05). Some samples from school A, D and E showed over 3.4 log CFU/100 $cm^2$ (total bacterial count) and 1.0 log CFU/100 $cm^2$ (total coliform), which requires immediate hygienic action. The number of customers per staff member, periodicity of hygiene education for staff and daily operation time of sterilizers were also found to be important factors related with the microbial contamination of food service operations. Conclusions: These results suggested that not only a HACCP (Hazard Analysis and Critical Control Point) approach, but also efforts to assess internal risk factors within operations be needed to reduce the microbial contamination of food utensils and fixtures. This study is expected to provide preliminary data for assessing microbial hazards in food service operations.

Probiotic Potential of Plant-Derived Lactic Acid Bacteria with Antihypertensive Activity (항고혈압 활성을 가진 식물유래 젖산균의 생균제 특성)

  • Lee, Ye-Ram;Son, Young-Jun;Park, Soo-Yun;Jang, Eun-Young;Yoo, Ji-Yeon;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.25 no.6
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    • pp.789-798
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    • 2016
  • Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

Lung Microbiome Analysis in Steroid-Naïve Asthma Patients by Using Whole Sputum

  • Jung, Jae-Woo;Choi, Jae-Chol;Shin, Jong-Wook;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung Whui;Park, Heung-Woo;Cho, Sang-Heon;Kim, Kijeong;Kang, Hye-Ryun
    • Tuberculosis and Respiratory Diseases
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    • v.79 no.3
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    • pp.165-178
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    • 2016
  • Background: Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid $na{\ddot{i}}ve$ asthmatics. Methods: Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-$na{\ddot{i}}ve$ asthma patients were analyzed through T-RFLP analysis. Results: In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae, and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls. Conclusion: Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls.

Microbial and Physicochemical Properties of Liquid Egg during Cold Storage (액란의 냉장저장 중 미생물 및 이화학적 성상)

  • Kang, Geun-Ho;Cho, Soo-Hyun;Seong, Pil-Nam;Park, Beom-Young;Ham, Jun-Sang;Jeong, Seok-Geun;Kim, Dong-Hun;Chae, Hyun-Seok
    • Food Science of Animal Resources
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    • v.31 no.4
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    • pp.557-562
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    • 2011
  • The study was performed to investigate microbial and physicochemical properties of domestic liquid eggs during cold storage. The liquid eggs used in the experiment were whole liquid, liquid egg yolks, and liquid egg whites. All samples were analyzed in summer and winter. The aerobic microorganisms were 1,270-83,300 CFU/g from non-sterilized liquid eggs produced in summer and their numbers increased from those produced in winter (ND, ~4,330 CFU/g). Total coliforms were not observed in non-sterilized whole liquid and non-sterilized liquid egg yolk regardless of season. Total coliforms from nonsterilized products were not detected in liquid egg whites during cold storage. Salmonella sp. was not observed in any of the liquid egg products. However, Pseudomonas sp., Pseudomonas geezennei, Pseudomonas otitidis, and Pseudomonas aeruginosa were identified by 16S rRNA from non-sterilized whole liquid eggs produced in summer. The pH and viscosity of whole liquid eggs and liquid egg whites were not different between the sterilized and non-sterilized treatments during cold storage. These results suggest that managing cross-contamination is necessary when non-sterilized liquid eggs are processed in summer.

Antioxidant Activity of Lactic Acid Bacteria Isolated from Korean Traditional Food Kimchi (한국전통식품 김치로부터 분리한 유산균주의 항산화 활성)

  • Kim, Da-Young;Kim, Hong Seok;Yoo, Jung Sik;Cho, Yoon Ah;Kim, Cheol-Hyun
    • Journal of Dairy Science and Biotechnology
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    • v.38 no.2
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    • pp.89-98
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    • 2020
  • The purpose of this study was to investigate the probiotic properties of lactic acid bacteria (LAB) isolated from a Korean traditional food kimchi. Gram staining was performed by Macrogen (Macrogen, Inc.) for identification of the LAB. Five strains of LAB were identified, including DKGF9 (Lactobacillus plantarum), DKGF1 (L. paracasei ), DKGF8 (L. casei ), DK207 (L. casei ), and DK211 (L. casei ). The biological activities of the isolated strains were assessed. The results showed that heat resistance of the strains was similar to or higher than the commercial strain L. acidophilus LA-5. Indirect testing of the ability of the strains to attach to the mucin layer revealed that DKGF9, DKGF1, and DKGF8 have high binding affinities for the mucous layer. All strains showed antimicrobial activity similar to or higher than the commercial strain LA-5. In proteolysis experiments, the diameters of proteolysis zones of the five strains increased in the period of 24-72 h, with DKGF1 exhibiting the largest zone diameter. Three strains were selected based on their antioxidant activities. Among the five isolated strains, L. paracasei DKGF1 showed potential probiotic activity, and thus, it may be useful for the development of health-promoting products.

Isolation and Characterization of Bacillus spp. with High-Level Productivity of Poly-γ-Glutamic Acid (Poly-γ-Glutamic Acid 고생성 Bacillus spp. 균주의 분리 및 발효특성)

  • Sim, SangHyeob;Park, Hong-Jin;Oh, HyeonHwa;Jeong, Do-Youn;Song, Geun-Seoup;Kim, Young-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.9
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    • pp.1114-1121
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    • 2017
  • Bacillus strains not producing harmful components were isolated from Korean traditional soybean products. Extracellular enzyme activities (amylase, protease, cellulase, and xylanase) of isolated Bacillus strains were measured, and Bacillus strains with high protease activity were selected. The selected 15 strains were identified as Bacillus amyloliquefaciens (10), Bacillus methylotrophicus (1), Bacillus velezensis (1), and Bacillus subtilis (3). Among them, B. subtilis JBG17019, B. amyloliquefaciens JBD17076, and B. amyloliquefaciens JBD17109 showed antimicrobial activities against food-borne microorganisms. The production abilities of glutamate, glutamine, and poly-${\gamma}$-glutamic acid (${\gamma}$-PGA) of the selected Bacillus strains were measured to analyze fermentation characteristics related to glutamic acid metabolism. The factor for multivariate was analyzed by the principal components analysis (PCA) method between fermentation characteristics and ${\gamma}$-PGA production. The three principal components were classified according to the PCA method: PC1 [enzyme activity (amylase, cellulase, and xylanase)], PC2 (${\gamma}$-PGA), and PC3 (protease, glutamate, and glutamine). As a result, B. amyloliquefaciens JBD17076 and B. subtilis JBG17019 strains were evaluated as having excellent enzyme activity and ${\gamma}$-PGA production.

Diversity of bacterial community during ensiling and subsequent exposure to air in whole-plant maize silage

  • Hu, Zongfu;Chang, Jie;Yu, Jianhua;Li, Shuguo;Niu, Huaxin
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.9
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    • pp.1464-1473
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    • 2018
  • Objective: To describe in-depth sequencing, the bacterial community diversity and its succession during ensiling of whole-plant maize and subsequent exposure to air. Methods: The microbial community dynamics of fermented whole-plant maize for 60 days (sampled on day 5, 10, 20, 40, 60) and subsequent aerobic exposure (sampled on day 63 after exposure to air for 3 days) were explored using Illumina Miseq sequence platform. Results: A total of 227,220 effective reads were obtained. At the genus level, there were 12 genera with relative abundance >1%, Lactobacillus, Klebsiella, Sporolactobacillus, Norank-c-cyanobacteria, Pantoea, Pediococcus, Rahnella, Sphingomonas, Serratia, Chryseobacterium, Sphingobacterium, and Lactococcus. Lactobacillus consistently dominated the bacterial communities with relative abundance from 49.56% to 64.17% during the ensiling process. Klebsiella was also an important succession bacterium with a decrease tendency from 15.20% to 6.41% during the ensiling process. The genus Sporolactobacillus appeared in late-ensiling stages with 7.70% abundance on day 40 and 5.32% on day 60. After aerobic exposure, the Lactobacillus decreased its abundance from 63.2% on day 60 to 45.03% on d 63, and Klebsiella from 5.51% to 5.64%, while Sporolactobacillus greatly increased its abundance to 28.15%. These bacterial genera belong to 5 phyla: Firmicutes (relative abundance: 56.38% to 78.43%) was dominant, others were Proteobacteria, Bacteroidetes, Cyanobacteria, and Actinobacteria. The bacterial communities clearly clustered into early-ensiling (d 5), medium-ensiling (d 10, d 20), late-ensiling (d 40, d 60), and aerobic exposure (d 63) clusters, with early- and late-ensiling communities more like each other than to the aerobic exposure communities. Conclusion: High-throughput sequencing based on 16S rRNA genes proved to be a useful method to explore bacterial communities of silage. The results indicated that the bacterial communities varied during fermentation and more dramatically during aerobic exposure. The study is valuable for understanding the mechanism of population change and the relationship between bacteria and ensilage characteristics.

Comparison of bacterial communities in leachate from decomposing bovine carcasses

  • Yang, Seung Hak;Ahn, Hee Kwon;Kim, Bong Soo;Chang, Sun Sik;Chung, Ki Yong;Lee, Eun Mi;Ki, Kwang Seok;Kwon, Eung Gi
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1660-1666
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    • 2017
  • Objective: Burial is associated with environmental effects such as the contamination of ground or surface water with biological materials generated during the decomposition process. Therefore, bacterial communities in leachates originating from the decomposing bovine carcasses were investigated. Methods: To understand the process of bovine (Hanwoo) carcass decomposition, we simulated burial using a lab-scale reactor with a volume of $5.15m^3$. Leachate samples from 3 carcasses were collected using a peristaltic pump once a month for a period of 5 months, and bacterial communities in samples were identified by pyrosequencing of the 16S rRNA gene. Results: We obtained a total of 110,442 reads from the triplicate samples of various sampling time points (total of 15 samples), and found that the phylum Firmicutes was dominant at most sampling times. Differences in the bacterial communities at the various time points were observed among the triplicate samples. The bacterial communities sampled at 4 months showed the most different compositions. The genera Pseudomonas and Psychrobacter in the phylum Proteobacteria were dominant in all of the samples obtained after 3 months. Bacillaceae, Clostridium, and Clostridiales were found to be predominant after 4 months in the leachate from one carcass, whereas Planococcaceae was found to be a dominant in samples obtained at the first and second months from the other two carcasses. The results showed that potentially pathogenic microbes such as Clostridium derived from bovine leachate could dominate the soil environment of a burial site. Conclusion: Our results indicated that the composition of bacterial communities in leachates of a decomposing bovine shifted continuously during the experimental period, with significant changes detected after 4 months of burial.

Microbial Floral Dynamics of Chinese Traditional Soybean Paste (Doujiang) and Commercial Soybean Paste

  • Gao, Xiuzhi;Liu, Hui;Yi, Xinxin;Liu, Yiqian;Wang, Xiaodong;Xu, Wensheng;Tong, Qigen;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1717-1725
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    • 2013
  • Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

  • Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.152-164
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    • 2015
  • Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.