• Korean Journal of Food Science and Technology
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• v.43 no.5
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• pp.611-617
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• 2011

This study investigated the germination and reduction of microbial population in domestic (radish, Chinese cabbage, and vitamin) and imported (radish and red cabbage) sprout seeds by heat treatment (40, 50, 60, and 70$^{\circ}C$ for 15 min or 30 min). The germination ratio (define the ratio) was 45-97% at 24 h after treatment <60$^{\circ}C$ and was decreased at 70$^{\circ}C$. In domestic radish seed, total aerobic bacteria were decrease by approximately 1.71 log CFU/g after heat treatment at 70$^{\circ}C$ for 30 min and viable coliforms were decreased to under the detection limit at temperatures over 60$^{\circ}C$. Decrease of total aerobic bacteria and coliforms in domestic Chinese cabbage seed was 1.23-1.34 and 2.77 log CFU/g, respectively, after heat treatment over 60$^{\circ}C$. In domestic vitamin seed, total aerobic bacteria were decreased by about 0.3 log CFU/g at 70$^{\circ}C$ for 15 min. In imported radish seed, total aerobic bacteria were decreased 2.12-2.30 log CFU/g after heat treatment over 60$^{\circ}C$. Total aerobic bacteria in imported red cabbage seed were reduced by 0.66-0.84 log CFU/g after heat treatment over 40$^{\circ}C$ and coliforms were undetectable. In case of Bacillus cereus, there was no significant difference by heat treatment in any sample. Staphylococcus aureus and Salmonella sp. were not detected at the detection limit in any tested seeds at any temperature.

• Molecular & Cellular Toxicology
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• v.2 no.1
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• pp.11-14
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• 2006

In this study we attempted to develop a novel genomic method to selectively isolate target functional microbial genomes from environmental samples. For this purpose, stable isotope probing (SIP) was applied in selectively isolating organic pollutant-assimilating populations. When soil microbes were fed with $^{13}C-labeled $ biphenyl, biphenyl-utilizing cells were incorporated with the heavy carbon isotope. The heavy DNA portion was successfully separated by CsCl equilibrium density gradient. And the diversity in the heavy DNA was sufficiently reduced, being suitable for the current DNA microarray techniques to detect biphenyl-utilizing populations in the soil. In addition, we proposed a new way to get more genetic information by combining this SIP method with selective metagenomic approach. The increased selective power of these new DNA isolation methods will be expected to provide a good quality of new genetic information, which, in turn, will result in development of a variety of biomarkers that may be used in assessing ecotoxicology issues including the impacts of organic hazards, and antibiotic-resistant pathogens on human and ecological systems.

• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1257-1267
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• 2023

Although rational genetic engineering is nowadays the favored method for microbial strain improvement, building up mutant libraries based on directed evolution for improvement is still in many cases the better option. In this regard, the demand for precise and efficient screening methods for mutants with high performance has stimulated the development of biosensor-based high-throughput screening strategies. Genetically encoded biosensors provide powerful tools to couple the desired phenotype to a detectable signal, such as fluorescence and growth rate. Herein, we review recent advances in engineering several classes of biosensors and their applications in directed evolution. Furthermore, we compare and discuss the screening advantages and limitations of two-component biosensors, transcription-factor-based biosensors, and RNA-based biosensors. Engineering these biosensors has focused mainly on modifying the expression level or structure of the biosensor components to optimize the dynamic range, specificity, and detection range. Finally, the applications of biosensors in the evolution of proteins, metabolic pathways, and genome-scale metabolic networks are described. This review provides potential guidance in the design of biosensors and their applications in improving the bioproduction of microbial cell factories through directed evolution.

• 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.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.1-6
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• 1999

A piezoelectric (PZ) biosensor system detecting Salmonella spp. was developed. The system consisted of an oscillator, a frequency counter and an antibody-immobilized quartz crystal. An anti-Salmonella antibody was immobilized on one gold. surface of the quartz crystal with protein A. Salmonella detection was made by measuring resonant frequency shift owing to a mass change by specific binding of microbial cells to the gold surface of the PZ crystal. The PZ antibody sensor was operated optimally at 0.1M phosphate buffer, pH 7.2 and $35^{\circ}C$. The sensor was quite specific to Salmonella spp. The obtained frequency shift was correlated with the Salmonella concentration in the range of $10^5{\sim}10^6\;CFU/mL$. The frequency shift increased further by addition of polystyrene beads. The Salmonella detection which is indicated by a steady-state microbial adsorption to the quartz crystal was accomplished within 50min.

• KSBB Journal
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• v.29 no.5
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• pp.361-371
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• 2014

Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants of biologic manufacturing processes. Conventional methods for mycoplasma testing, direct culture method and indirect indicator cell culture method, are lengthy, costly and less sensitive to noncultivable species. In this report, we describe a new TaqMan probe-based real-time PCR method for rapid and quantitative detection of mycoplasma contamination during manufacture of biologics. Universal mycoplasma primers were used for mycoplasma PCR and mycoplasma DNA was quantified by use of a specific TaqMan probe. Specificity, sensitivity, and robustness of the real-time PCR method was validated according to the European Pharmacopoeia. The validation results met required criteria to justify its use as a replacement for the culture method. The established real-time PCR assay was successfully applied to the detection of mycoplasma from human keratinocyte and mesenchymal stem cell as well as Vero cell lines artificially infected with mycoplasma. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of mycoplasma contamination during manufacture of biologics.

• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.465-470
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• 2016

Sample preparation methods were evaluated for effectiveness in detecting foodborne pathogens from sprout seeds. The methods included: Rinse.-Test portions were rinsed with 0.1% peptone water, and the pellet after centrifugation was inoculated into pre-enrichment media; and Sprouting.-Seed samples were sprouted before pre-enrichment and sprouted seeds were inoculated into pre-enrichment media. In rinse method, E. coli was isolated from 13 of 280 sample units. In sprouting method, E. coli was isolated from 12 of 135 sample units. E. coli O157:H7, Salmonella spp., and L. monocytogenes were not detected in any of the samples. In the trials for recovering Salmonella enterica from artificially contaminated alfalfa seeds, the soak, rinse, and sprouting methods were evaluated. The detection rates of S. enterica were statistically different according to the amount of the sample tested and selective medium type (P < 0.05).

• Journal of Biosystems Engineering
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• v.39 no.3
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• pp.215-226
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• 2014

Recently, spectroscopy has emerged as a potential tool for quality evaluation of numerous food and agricultural products because it provides information regarding both spectral distribution and image features of the sample (i.e., hyperspectral imaging). Spectroscopic techniques reveal hidden information regarding the sample and do so in a non-destructive manner. This review describes the various approaches of spectroscopic modalities, especially hyperspectroscopy and vibrational spectroscopies (i.e., Raman spectroscopy and Fourier transform near infrared spectroscopy) combined with chemometrics for the non-destructive assessment of contaminations and defects in agro-food products.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.129-138
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• 1999

If rumen bacteria can be manipulated to utilize nutrients (i.e., ammonia and plant cell wall carbohydrates) more completely and efficiently, the need for protein supplementation can be reduced or eliminated and the digestion of fiber in forage or agricultural residue-based diets could be enhanced. However, these approaches require a complete and accurate description of the rumen community, as well as methods for the rapid and accurate detection of microbial density, diversity, phylogeny, and gene expression. Molecular ecology techniques based on small subunit (SSU) rRNA sequences, nucleic acid probes and the polymerase chain reaction (PCR) can potentially provide a complete description of the microbial ecology of the rumen of ruminant animals. The development of these molecular tools will result in greater insights into community structure and activity of gut microbial ecosystems in relation to functional interactions between different bacteria, spatial and temporal relationships between different microorganisms and between microorganisms and reed panicles. Molecular approaches based on SSU rRNA serve to evaluate the presence of specific sequences in the community and provide a link between knowledge obtained from pure cultures and the microbial populations they represent in the rumen. The successful development and application of these methods promises to provide opportunities to link distribution and identity of gastrointestinal microbes in their natural environment with their genetic potential and in situ activities. The use of approaches for assessing pupulation dynamics as well as for assessing community functionality will result in an increased understanding and a complete description of the gastrointestinal communities of production animals fed under different dietary regimes, and lead to new strategies for improving animal growth.

• Food Science of Animal Resources
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• v.35 no.5
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• pp.674-682
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• 2015

The objective of this study was to evaluate the risk of illness from Campylobacter spp. on ham. To identify the hazards of Campylobacter spp. on ham, the general characteristics and microbial criteria for Campylobacter spp., and campylobacteriosis outbreaks were investigated. In the exposure assessment, the prevalence of Campylobacter spp. on ham was evaluated, and the probabilistic distributions for the temperature of ham surfaces in retail markets and home refrigerators were prepared. In addition, the raw data from the Korea National Health and Nutrition Examination Survey (KNHNES) 2012 were used to estimate the consumption amount and frequency of ham. In the hazard characterization, the Beta-Poisson model for Campylobacter spp. infection was used. For risk characterization, a simulation model was developed using the collected data, and the risk of Campylobacter spp. on ham was estimated with @RISK. The Campylobacter spp. cell counts on ham samples were below the detection limit (<0.70 Log CFU/g). The daily consumption of ham was 23.93 g per person, and the consumption frequency was 11.57%. The simulated mean value of the initial contamination level of Campylobacter spp. on ham was −3.95 Log CFU/g, and the mean value of ham for probable risk per person per day was 2.20×10⁻¹². It is considered that the risk of foodborne illness for Campylobacter spp. was low. Furthermore, these results indicate that the microbial risk assessment of Campylobacter spp. in this study should be useful in providing scientific evidence to set up the criteria of Campylobacter spp..