• Title/Summary/Keyword: Microbiology Quality

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Immunochromatographic Strip Assay for Detection of Cronobacter sakazakii in Pure Culture

  • Song, Xinjie;Shukla, Shruti;Lee, Gibaek;Kim, Myunghee
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1855-1862
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    • 2016
  • Cronobacter sakazakii (C. sakazakii) is a foodborne pathogen, posing a high risk of disease to infants and immunocompromised individuals. In order to develop a quick, easy, and sensitive assay for detecting C. sakazakii, a rabbit anti-C. sakazakii immunoglobulin G (IgG) was developed using sonicated cell protein from C. sakazakii. The developed anti-C. sakazakii (IgG) was of good quality and purity, as well as species-specific. The developed rabbit anti-C. sakazakii IgG was attached to the surface of a sulforhodamine B-encapsulated liposome to form an immunoliposome. A test strip was then prepared by coating goat anti-rabbit IgG onto the control line and rabbit anti-C. sakazakii IgG onto the test line, respectively, of a plastic-backed nitrocellulose membrane. A purple color signal both on the test line and the control line indicated the presence of C. sakazakii in the sample, whereas purple color only on the control line indicated the absence of C. sakazakii in the sample. This immunochromatographic strip assay could produce results in 15 min with a limit of detection of $10^7CFU/ml$ in C. sakazakii culture. The immunochromatographic strip assay also showed very good specificity without cross-reactivity with other tested Cronobacter species. Based on these results, the developed immunochromatographic strip assay is efficient for the detection of C. sakazakii and has high potential for on-site detection.

Polyphasic Microbial Analysis of Traditional Korean Jeung-Pyun Sourdough Fermented with Makgeolli

  • Lim, Sae Bom;Tingirikari, Jagan Mohan Rao;Kwon, Ye Won;Li, Ling;Kim, Grace E.;Han, Nam Soo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.226-233
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    • 2017
  • Jeung-pyun, a fermented rice cake, is prepared by fermenting rice sourdough using makgeolli, a traditional Korean rice wine, in the presence of yeast and lactic acid bacteria (LAB). The goal of this study was to conduct biochemical and microbial analyses of five different rice sourdoughs, each fermented with a different commercial makgeolli, using culture-dependent and culture-independent approaches. All sourdough samples fermented with different makgeolli for 6.5 h showed different profiles in pH, total titratable acidity, organic acid concentration, and microbial growth. LAB belonging to different genera were identified based on colony morphology on modified MRS and sourdough bacteria agar medium. PCR-denaturing gradient gel electrophoresis analyses of the five sourdoughs showed different bands corresponding to LAB and yeast. 16S/26S rRNA gene sequence analyses of the samples confirmed that the predominant LAB in the five fermented rice doughs was Lactobacillus plantarum, Lb. pentosus, and Lb. brevis. Various other Lactobacillus spp. and Saccharomyces cerevisiae were common in all five fermented samples. This study provides comprehensive and comparative information on the microflora involved in fermentation of rice sourdough and signifies the need to develop effective starters to enrich the quality of jeung-pyun.

Combined Treatment on the Inactivation of Naturally Existing Bacteria and Escherichia coli O157:H7 Inoculated on Fresh-Cut Kale

  • Kang, Ji Hoon;Song, Kyung Bin
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.219-225
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    • 2017
  • An aqueous chlorine dioxide ($ClO_2$) treatment combined with highly activated calcium oxide (CaO) and mild heat was tested for inactivating naturally existing bacteria and Escherichia coli O157:H7 inoculated on fresh-cut kale. Kale samples were treated with different concentrations of $ClO_2$ (10, 30, and 50 ppm), CaO (0.01%, 0.05%, 0.1%, and 0.2%), and mild heat ($25^{\circ}C$, $45^{\circ}C$, $55^{\circ}C$, and $65^{\circ}C$) as well with combinations of 30 or 50 ppm $ClO_2$ and 0.2% CaO at $55^{\circ}C$ for 3 min. An increasing concentration of $ClO_2$ and CaO significantly reduced the microbial population compared with the control. In addition, mild heating at $55^{\circ}C$ elicited greater microbial reduction than the other temperatures. A combined treatment of 50 ppm $ClO_2$ and 0.2% CaO at $55^{\circ}C$ reduced the population of naturally existing bacteria on kale by 3.10 log colony forming units (CFU)/g, and the counts of E. coli O157:H7 were below the detection limit (1 log CFU/g). In addition, no significant differences in the Hunter color values were evident in any treatment during storage. Therefore, a combined treatment of $ClO_2$ and active CaO at $55^{\circ}C$ can be an effective sanitizing method to improve the microbiological safety of fresh-cut kale without affecting its quality.

Insights into the Gut Microbiota of Freshwater Shrimp and Its Associations with the Surrounding Microbiota and Environmental Factors

  • Zhao, Yanting;Duan, Cuilan;Zhang, Xu-xiang;Chen, Huangen;Ren, Hongqiang;Yin, Ying;Ye, Lin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.946-956
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    • 2018
  • The gut microbiota of aquatic animals plays a crucial role in host health through nutrient acquisition and outcompetition of pathogens. In this study, on the basis of the high-throughput sequencing of 16S rRNA gene amplicons, we examined the bacterial communities in the gut of freshwater shrimp (Macrobrachium nipponense) and in their living environments (sediment and pond water) and analyzed the effects of abiotic and biotic factors on the shrimp gut bacterial communities. High bacterial heterogeneity was observed in the freshwater shrimp gut samples, and the result indicated that both the surrounding bacterial community and water quality factors (particularly dissolved oxygen and temperature) could affect the shrimp gut bacterial community. Despite the observed heterogeneity, 57 genera, constituting 38-99% of the total genera in each of the 40 shrimp gut samples, were identified as the main bacterial population in the gut of M. nipponense. In addition, a high diversity and abundance of lactic acid bacteria (26 genera), which could play significant roles in the digestion process in shrimp, were observed in the shrimp gut samples. Overall, this study provides insights into the gut bacterial communities of freshwater shrimp and basic information for shrimp farming regarding the application of probiotics and disease prevention.

Microbial Community Structure of Korean Cabbage Kimchi and Ingredients with Denaturing Gradient Gel Electrophoresis

  • Hong, Sung Wook;Choi, Yun-Jeong;Lee, Hae-Won;Yang, Ji-Hee;Lee, Mi-Ai
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1057-1062
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    • 2016
  • Kimchi is a traditional Korean fermented vegetable food, the production of which involves brining of Korean cabbage, blending with various other ingredients (red pepper powder, garlic, ginger, salt-pickled seafood, etc.), and fermentation. Recently, kimchi has also become popular in the Western world because of its unique taste and beneficial properties such as antioxidant and antimutagenic activities, which are derived from the various raw materials and secondary metabolites of the fermentative microorganisms used during production. Despite these useful activities, analysis of the microbial community present in kimchi has received relatively little attention. The objective of this study was to evaluate the bacterial community structure from the raw materials, additives, and final kimchi product using the culture-independent method. Specifically, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the 16S rRNA partial sequences of the microflora. One primer set for bacteria, 341FGC-518R, reliably produced amplicons from kimchi and its raw materials, and these bands were clearly separated on a 35-65% denaturing gradient gel. Overall, 117 16S rRNA fragments were identified by PCR-DGGE analysis. Pediococcus pentosaceus, Leuconostoc citreum, Leuconostoc gelidum, and Leuconostoc mesenteroides were the dominant bacteria in kimchi. The other strains identified were Tetragenococcus, Pseudomonas, Weissella, and uncultured bacterium. Comprehensive analysis of these microorganisms could provide a more detailed understanding of the biologically active components of kimchi and help improve its quality. PCR-DGGE analysis can be successfully applied to a fermented food to detect unculturable or other species.

Evaluation of Yeast Diversity During Wine Fermentations with Direct Inoculation and pied de cuve Method at an Industrial Scale

  • Li, Erhu;Liu, Chuanhe;Liu, Yanlin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.960-966
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    • 2012
  • The diversity and composition of yeast populations may greatly impact wine quality. This study investigated the yeast microbiota in two different types of wine fermentations: direct inoculation of a commercial starter versus pied de cuve method at an industrial scale. The pied de cuve fermentation entailed growth of the commercial inoculum used in the direct inoculation fermentation for further inoculation of additional fermentations. Yeast isolates were collected from different stages of wine fermentation and identified to the species level using Wallersterin Laboratory nutrient (WLN) agar followed by analysis of the 26S rDNA D1/D2 domain. Genetic characteristics of the Saccharomyces cerevisiae strains were assessed by a rapid PCR-based method, relying on the amplification of interdelta sequences. A total of 412 yeast colonies were obtained from all fermentations and eight different WL morphotypes were observed. Non-Saccharomyces yeast mainly appeared in the grape must and at the early stages of wine fermentation. S. cerevisiae was the dominant yeast species using both fermentation techniques. Seven distinguishing interdelta sequence patterns were found among S. cerevisiae strains, and the inoculated commercial starter, AWRI 796, dominated all stages in both direct inoculation and pied de cuve fermentations. This study revealed that S. cerevisiae was the dominant species and an inoculated starter could dominate fermentations with the pied de cuve method under controlled conditions.

Useful and Effective Diagnosis and Evaluation Tools for Eenvironmental Change in Increased Mill Water System Closure

  • Linda R. Robertson;Lee, Byung-Tae;Kim, Tae-Joon
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.33 no.5
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    • pp.1-11
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    • 2001
  • In the past, abundant and clean water was available for paper mills'use. However, the growth of population and industry made water less available nowadays. Also, environmental regulation limits wastewater discharge, which affects mill operation cost. Therefore, paper mills are under pressure to use more recycled water and mill system closure. As a result, chemical and physical parameters of water are changing and new environment if being created for microorganisms in paper mill system as well. The more soluble or suspended organic materials are increased as more water is recycled and less or scarce dissolved oxygen is available, depending on the degree of recycled water usage. Microorganism flora ill paper mill system will be a1so shifted according to the environmental change of mill system. Anaerobic bacteria, including sulfate reducing bacteria (SRB), will be dominant in the system as very low or almost no oxygen available in the system. Nevertheless, it is common in domestic paper mills that employ the same and old biocides as a means of microbial control, and microbiological control is often less recognized or even neglected. The right biocide selection for increased reductive environment of mills is critical for operation and estimated loss from paper quality defects such as sheet break, holes due to microbiological cause is tremendous compared to the microbiological control cost. It is imperative to investigate and diagnosis the environmental change of mills for right control of cumbersome microorganisms. Several useful diagnosis tools, including new technology employing OFM(Optical Fouling Monitor) in situ, are illustrated.

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Physicochemical Properties and Bacterial Communities of Meongge (Halocynthia roretzi) Jeotgal Prepared with 3 Different Types of Salts

  • Kim, Jeong A;Yao, Zhuang;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.527-537
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    • 2019
  • Three types of meongge (Halocynthia roretzi) jeotgal (MJ) were prepared with 3 different types of salts (12%, w/v): purified salt (PS), solar salt aged for 3 years (SS), and bamboo salt that had been recrystalized 3 times (BS). One set of MJ was fermented with starters, Bacillus subtilis JS2 and Tetragenococcus halophilus BS1-37 (each 6 log CFU/g), and another set without starters for 42 days at $10^{\circ}C$. The LAB count of the SSMJ (non-starter) was highest at day 28 (2.30 log CFU/g). The pH of the PSMJ and SSMJ was 5.72-5.77 at day 0, and 5.40-5.50 at day 42. BSMJ showed higher pH and lower titratable acidities than other samples. Amino-type nitrogen (ANN) increased continuously, and SSMJ showed higher values than other samples from day 14. Bacterial species of non-starter MJ were examined by culture independent method. Clone libraries of 16S rRNA genes were constructed in Escherichia coli from total DNA from non-starter MJ samples at day 0, 14, and 28. Thirty clones per each sample were randomly selected and DNA sequences were analyzed. Variovorax sp., uncultured bacterium, and Acidovorax sp. were the most dominant group at day 0, 14, and 28, respectively. Lactobacillus sakei and Streptococcus sp. were the next dominant group in SSMJ at day 28. A Streptococcus sp. was detected from PSMJ at day 28. Sensory evaluation for MJ samples at day 28 showed that SSMJ got higher overall acceptability scores. These results showed that solar salt can cause desirable changes in the microbial community of fermented foods, thereby positively affecting their overall quality.

Nano-Encapsulation of Plant Growth-Promoting Rhizobacteria and Their Metabolites Using Alginate-Silica Nanoparticles and Carbon Nanotube Improves UCB1 Pistachio Micropropagation

  • Pour, Mojde Moradi;Saberi-Riseh, Roohallah;Mohammadinejad, Reza;Hosseini, Ahmad
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1096-1103
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    • 2019
  • UCB-1 is the commercial rootstock of pistachio. Reproduction of this rootstock by tissue culture is limited by low levels of proliferation rate. Therefore, any compound that improves the proliferation rate and the quality of the shoots can be used in the process of commercial reproduction of this rootstock. Use of plant growth-promoting bacteria is one of the best ideas. Given the beneficial effects of nanoparticles in enhancement of the growth in plant tissue cultures, the aim of the present study was to investigate the effects of nanoencapsulation of plant growth-promoting rhizobacteria (using silica nanoparticles and carbon nanotubes) and their metabolites in improving UCB1 pistachio micropropagation. The experiment was conducted in a completely randomized design with three replications. Before planting, treatments on the DKW medium were added. The results showed that the use of Pseudomonas fluorescens VUPF5 and Bacillus subtilis VRU1 nanocapsules significantly enhanced the root length and proliferation. The nanoformulation of the VUPF5 metabolite led to the highest root length (6.26 cm) and the largest shoot (3.34 cm). Inoculation of explants with the formulation of the metabolites (both bacterial strains) significantly elevated the average shoot length and the fresh weight of plant compared to the control. The explants were dried completely using both bacterial strains directly and with capsule coating after the three days.

Semi-Rational Screening of Probiotics from the Fecal Flora of Healthy Adults against DSS-Induced Colitis Mice by Enhancing Anti-Inflammatory Activity and Modulating the Gut Microbiota

  • Wang, Weiwei;Xing, Wentao;Wei, Sichen;Gao, Qiaoying;Wei, Xinliang;Shi, Liang;Kong, Yu;Su, Zhenhua
    • Journal of Microbiology and Biotechnology
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    • v.29 no.9
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    • pp.1478-1487
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    • 2019
  • Ulcerative colitis (UC), a chronic inflammatory bowel disease, substantially impacts patients' health-related quality of life. In this study, an effective strategy for discovering high-efficiency probiotics has been developed. First, in order to survive in the conditions of the stomach and intestine, high bile salt-resistant and strong acid-resistant strains were screened out from the fecal flora of healthy adults. Next, the probiotic candidates were rescreened by examining the induction ability of IL-10 (anti-inflammatory factor) production in dextran sodium sulfate (DSS)-induced colitis mice, and Lactobacillus sakei 07 (L07) was identified and selected as probiotic P. In the end, fourteen bifidobacterium strains isolated from stools of healthy males were examined for their antimicrobial activity. Bifidobacterium bifidum B10 (73.75% inhibition rate) was selected as probiotic B. Moreover, the colonic IL-6 and $TNF-{\alpha}$ expression of the DSS-induced colitis mice treated with L. sakei 07 (L07) - B. bifidum B10 combination (PB) significantly decreased and the IL-10 expression was up-regulated by PB compared to the DSS group. Furthermore, Bacteroidetes and Actinobacteria decreased and Firmicutes increased in the DSS group mice, significantly. More interestingly, the intestinal flora biodiversity of DSS colitis mice was increased by PB. Of those, the level of B. bifidum increased significantly. The Bacteriodetes/Firmicutes (B/F) ratio increased, and the concentration of homocysteine and LPS in plasma was down-regulated by PB in the DSS-induced colitis mice. Upon administration of PB, the intestinal permeability of the the DSS-induced colitis mice was decreased by approximately 2.01-fold. This method is expected to be used in high-throughput screening of the probiotics against colitis. In addition, the L. sakei 07 - B. bifidum B10 combination holds potential in UC remission by immunomodulatory and gut microbiota modulation.