• Korean Chemical Engineering Research
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• v.61 no.2
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• pp.247-257
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• 2023

Precise counting of cell number stands in important position within clinical and research laboratories. Conventional methods such as hemocytometer, migration/invasion assay, or automated cell counters have limited in analytical time, cost, and accuracy., which needs an alternative way with time-efficient in-situ approach to broaden the application avenue. Here, we present simple coding-based cell counting method using image analysis tool, freely available image software (ImageJ). Firstly, we encapsulated RFP-expressing bacteria in a droplet using microfluidic device and automatically performed fluorescence image-based analysis for the quantification of cell numbers. Also, time-lapse images were captured for tracking the change of cell numbers in a droplet containing different concentrations of antibiotics. This study confirms that our approach is approximately 15 times faster and provides more accurate number of cells in a droplet than the external analysis program method. We envision that it can be used to the development of high-throughput image-based cell counting analysis.

• Korean Journal of Food Science and Technology
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• v.33 no.4
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• pp.461-468
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• 2001

The effectiveness and safety of Enzyme Food, a group of dietary supplements designated by Korean Food Law, were evaluated and the possibility of HACCP (Hazard Analysis Critical Control Point) application was investigated. Chemical composition, enzyme activities and the degree of bacterial contamination in 12 samples of different brands sold in Korean market were measured. The chemical composition of the selected products varied and inconsistent to those claimed in the label description. It is known that effectiveness of Enzyme Food depends on enzyme activity, but enzyme activities of ${\alpha}-amylase$ varied from $1,793\;{\mu}g/min$ g to $159\;{\mu}g/min$ g and those of ${\beta}-amylase$ ranged from $171\;{\mu}g/min$ g to $11\;{\mu}g/min$ g. The protease activities varied from $27.57\;{\mu}g/min$ g to $0.18\;{\mu}g/min$ g. In coli-form bacterial test, positive reactions were appeared in the 50 % of the samples. Numbers of bacteria ranged from $1.3\;{\times}\;10^5\;to\;1.2\;{\times}\;10^9$. Five CCPs were identified; heating, inoculation, cultivation, drying and granulation. Consideration of HACCP system indicated that the pretreatment of raw material, checking of bacterial contamination and stability of enzyme activity during fermentation process were important factors for the quality of Enzyme Foods.

• Korean Journal of Soil Science and Fertilizer
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• v.50 no.3
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• pp.135-149
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• 2017

In order to obtain promising rice growth-promoting microbial strains that can be used as substitutes for chemical fertilizers, 172 bacterial strains were isolated from rice roots grown in Korean and Russian soils. Out of them, the strains KR076, KR083, KR181 and RRj228 showed plant growth-promoting activities on maize seedlings. Bacillus megaterium KR076 and Bacillus sp. KR083 showed both nitrogen-fixing and plant growth-promoting activities, while Rhizobium sp. KR181 and Pseudomonas sp. RRj228 appeared to support only plant growth-promotion, but not $N_2$ fixation. Especially, RRj228 showed high growth promoting activity at low concentrations. Inoculation studies with KR083 and RRj228 revealed a high affinity to the Japonica rice variety such as Junambyeo than the Korean Tongil type variety such as Arumbyeo. Both KR083 and RRj228 strains showed rhizoplane and/or endophytic colonization in Japonica and Tongil types rice when soaked with the bacterial suspension of $1.1{\times}10^5cfu\;ml^{-1}$ for six and twelve hours. However, the total bacterial cell numbers were higher in the roots of Japonica variety than in the Tongil type. In inoculation trials with Daesanbyeo rice variety, the seedlings inoculated with KR181 and RRj228 at the rate of $2.0{\times}10^6cfu\;ml^{-1}$ showed yield increment of 35% and 33% (p < 0.01), respectively, so that they contributed to the replacement of chemical fertilizer at half doses of N, $P_2O_5$, and $K_2O$ in pots. In Junambyeo rice seedlings, the strain RRj228, when inoculated with a cell suspension of $1.8{\times}10^6cfu\;ml^{-1}$, promoted 3.4% higher yield at 70% dose than at a full dose level of N $110kg\;ha^{-1}$ in field. These results suggest that the rhizobacteria KR181 and RRj228 are prospective strains for enhancing rice performance.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.12 no.1
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• pp.19-26
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• 1979

To evaluate the sanitary conditions of four species of commercial shellfish (Crassostrea gigas, Tapes japonica, Mytilus coruscus and Anadara granosa), the psychrotrophic coliforms and the bacterial flora were monthly examined from January to December, 1978. Viable cell counts of the samples submitted were $1.3\times10^3\~2.5\times10^6/g$ and the numbers of coliform and E. coli were above $0\~5.5\times10^2/g\;and\;2\~9.2\times10^2/100g$ respectively. Among the 373 coliform bacteria isolated from the samples submitted, 298 strains were identified as coliform by IMViC test, $44^{\circ}C$ culture and gelatin liquefaction test. E. coli type I and K. aerogenes type I were the most predominant ones among the identified coliform as 138 strains and 71 strains respectively. The psychrotrophic coliform growing at $5^{\circ}C$ were 186 $strains(50\%)$ among 373 strains of isolated coliform. Vibrio and Pseudomonas were the most predominant groups. Among 453 strains of isolated bacteria, Vibrio was 240 $strains(53\%)$ and Pseudomonas was 91 $strains(20\%)$. Moraxella, Acinetobacter, Enterobacteriaceae and Flavobacterium-Cytophage were scantly detected. In seasonal variation of general bacterial flora, a lot of Vibrio appeared from May to October, did Moraxella during the winter season and the other genuses showed no seasonal change.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.4
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• pp.615-620
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• 2013

To extend the shelf-life of salted Chinese cabbages, electrolyzed water (EW) was used to wash raw Chinese cabbages before the salting process (to control microbial growth), and different storage temperatures evaluated (0, 4, and $10^{\circ}C$). A tap water washing group (TW) was used for comparison. The initial total bacterial population was 5.36 log CFU/g in the TW treatment and 3.50 log CFU/g in the EW treatment. The EW treatment decreased bacterial numbers by approximately 2 log CFU/g compared to the TW treatment and kept this initial population number for 32 days at $0^{\circ}C$. The salinity had no difference during storage. In general, several factors (pH, reducing sugars, hardness, and transmittance) decreased over time, and decreased slowly with EW treatment and $0^{\circ}C$ storage. Overall, the salted Chinese cabbages with EW treatment showed lower bacterial populations compared to TW treatment, and when stored at $0^{\circ}C$, delayed decreases in quality.

• Korean Journal of Environmental Biology
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• v.25 no.3
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• pp.273-278
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• 2007

The cell numbers of heterotrophic bacteria inhabiting the surface and bottom sea water harvested from the 5 stations in the marine ranching ground of Tongyeong coastal waters in $2003{\sim}2007$ were examined, and species composition of the heterotrophic bacterial population and dominant species were analyzed as well. Sea water samples collected in summer season contained much higher number of heterotrophic bacteria than those harvested in winter, spring and autumn seasons due to the higher sea water temperature. However the cell number of heterotrophic bacteria did not show a significant dependence on the location of the sampling stations. The cell number of heterotrophic bacteria in the surface sea water harvested in October 2003 and in September 2004 was not discernibly different from that in the bottom sea water and sometimes the former was even fewer than the latter because of the typhoon and localized torrential downpour. The number of heterotrophic bacteria decreased every year. The main bacterial species were Pseudomonas fluorescens TY1, Pseudomonas stutzeri TY2, Acinetobacter lwoffii TY3, Sphingomonas paucimobilis TY4, Burkholderia mallei TY5, Pasteurella haemolytica TY6, Pasteurella multocida TY7, Comamonas acidovorans TY8, Actinobacillus ureae TY9 and Chryseobacterium indologenes TY10. P. fluorescens TY1 and A. lwoffii TY3 were found to be the dominant species.

• Korean Journal of Microbiology
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• v.38 no.1
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• pp.31-37
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• 2002

For elucidating the correlation between the eubacterial community structure and environmental parameters in Nammae Reservoir located in Kyungsan, Kyungbuk, the bacterial community structure and their structure affecting environmental parameters were analyzed using Fluorescent In Situ Hybridization (FISH) monthly over year. $\alpha$ . $\beta$ . $\gamma$-subclasses of Proteobacteria and Cytophaga-Flavobacterium (CF) group known as dominant bacterial group in freshwater were detected in 3 stations over year. The ratio of each subclass to total bacteria was determined; $\alpha$.$\beta$ . $\gamma$-subclasses and CF group varied in the range of 4.0~29.2%, 1.7~25.8%, 1.8~12.8%, 4.9~36.3%, respectively and there was no substantial differences between stations. In terms of the correlation between each group specific bacteria and environmental parameters such as temperature, SS, pH, DOC, NH$_4$-N, NO$_3$-N, PO$_4$-P, standing crops of algae, the results were as follows: 1) total bacterial numbers correlated positively with temperature, SS and DOC, 2) Eubacteria positively with DOC and Chl-$\alpha$, 3)${\gamma}$-subclass positively with DOC, and 4) CF group positively with standing crops of chlorophyceae, 5) whereas $\beta$-subclass bacteria correlated negatively with standing crop of cyanobacteria and that of total algae.

• Korean Journal of Microbiology
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• v.36 no.4
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• pp.292-298
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• 2000

For scrutinizing the changes of aggregated bacteria in Lake Paldang, the FISH method was applied by using the rhodamine labeled probes, and total bacteria, chloropyll a concentrations and nutrients concentrations were measured. The aggregates were collected with sediment traps. The T-N, T-P, chlorophyll a concentrations of aggregates were higher 5-15 times, 81-140 and 49-66 times than water samples, respectively. Also, the bacterial numbers of aggregates were 200 times higher than those of water smaples. The ratios of each groups of water sample were 2.1-7.4% for $\alpha$-group, 4.5-8.3% for $\beta$-group, 2.1-7.4% for $\gamma$-group, 2.1-6.1% for Cytophaga-Flavobacterium group and 0.1-2.5% for 'other'group, respectively. While, in aggregates, the ratios of $\alpha$-, $\beta$-, $\gamma$- and Cytophaga-Flavobacterium groups were very small and most abundant group was 'other' bacteria. With these results, the aggregated bacteria in Lake Paldang had a particular group composition of bacteria.

• Toxicological Research
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• v.35 no.1
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• pp.45-63
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• 2019

In view of the growing industrial use of Bacterial cellulose (BC), and taking into account that it might become airborne and be inhaled after industrial processing, assessing its potential pulmonary toxic effects assumes high relevance. In this work, the murine model was used to assess the effects of exposure to respirable BC nanofibrils (nBC), obtained by disintegration of BC produced by Komagataeibacter hansenii. Murine bone marrow-derived macrophages ($BMM{\Phi}$) were treated with different doses of nBC (0.02 and 0.2 mg/mL, respectively 1 and $10{\mu}g$ of fibrils) in absence or presence of 0.2% Carboxymethyl Cellulose (nBCMC). Furthermore, mice were instilled intratracheally with nBC or nBCMC at different concentrations and at different time-points and analyzed up to 6 months after treatments. Microcrystaline $Avicel-plus^{(R)}$ CM 2159, a plant-derived cellulose, was used for comparison. Markers of cellular damage (lactate dehydrogenase release and total protein) and oxidative stress (hydrogen peroxidase, reduced glutathione, lipid peroxidation and glutathione peroxidase activity) as well presence of inflammatory cells were evaluated in brochoalveolar lavage (BAL) fluids. Histological analysis of lungs, heart and liver tissues was also performed. BAL analysis showed that exposure to nBCMC or CMC did not induce major alterations in the assessed markers of cell damage, oxidative stress or inflammatory cell numbers in BAL fluid over time, even following cumulative treatments. $Avicel-plus^{(R)}$ CM 2159 significantly increased LDH release, detected 3 months after 4 weekly administrations. However, histological results revealed a chronic inflammatory response and tissue alterations, being hypertrophy of pulmonary arteries (observed 3 months after nBCMC treatment) of particular concern. These histological alterations remained after 6 months in animals treated with nBC, possibly due to foreign body reaction and the organism's inability to remove the fibers. Overall, despite being a safe and biocompatible biomaterial, BC-derived nanofibrils inhalation may lead to lung pathology and pose significant health risks.

• Journal of Marine Life Science
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• v.1 no.1
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• pp.18-24
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• 2016

Since April 2012, doctor fish in the breeding tank and in the quarantine tank in Hanwha Aquaplanet Yeosu Aquarium have been dying, accompanied by diffuse bleeding around the mouth, in the chin, and at the bottom of the abdomen. In this study, the cause of death would be examined through the bacteriological study of doctor fish and the rearing water quality in the aquarium. The water quality and the bacterial counts of the rearing water in the exhibit tank and in the quarantine tank were analyzed once a week, starting from August to November 2014. Water quality was measured based on the following data: temperature was in the range of 24.5~26.8℃, pH at 6.77~7.94, DO at 6.15~8.61 ppm, ammonia at 0~0.93 ppm, nitrite at 0.009~0.075 ppm, and nitrate at 1.1~40.9 ppm. Studies revealed that the differences in these water quality factors were not related to the death of doctor fish. Bacterial counts in the rearing waters of Garra rufa slightly increased to 10³~10⁴ CFU/ml, just before the death of the doctor fish. Twelve strains of bacteria were isolated from the dead fish and rearing waters. The isolates were identified as Aeromonas veronii, Citrobacter freundii, Pseudorhodoferax aquiterrae, Shewanella putrefaciens, and Vibrio anguillarum on the basis of 16S rRNA gene sequences. The most dominant species was C. freundii, which showed medium sensitivity to florfenicol and norfloxacin, and was resistant to amoxacillin, doxycycline, oxytetracycline, tetracycline, and trimethoprim. Ten isolates were confirmed to be pathogenic to the doctor fish. Doctor fish infected with C. freundii and S. putrefaciens showed high mortality in the experimental groups. These results indicate that the variation in bacterial numbers in the rearing water was related to the death of doctor fish. C. freundii and S. putrefaciens were directly implicated in causing the death of doctor fish in the aquarium.