• Food Science of Animal Resources
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• v.44 no.5
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• pp.1080-1095
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• 2024

In contemporary society, the increasing number of pet-owning households has significantly heightened interest in companion animal health, expanding the probiotics market aimed at enhancing pet well-being. Consequently, research into the gut microbiota of companion animals has gained momentum, however, ethical and societal challenges associated with experiments on intelligent and pain-sensitive animals necessitate alternative research methodologies to reduce reliance on live animal testing. To address this need, the Fermenter for Intestinal Microbiota Model (FIMM) is being investigated as an in vitro tool designed to replicate gastrointestinal conditions of living animals, offering a means to study gut microbiota while minimizing animal experimentation. The FIMM system explored interactions between intestinal microbiota and probiotics within a simulated gut environment. Two strains of commercial probiotic bacteria, Enterococcus faecium IDCC 2102 and Bifidobacterium lactis IDCC 4301, along with a newly isolated strain from domestic dogs, Lactobacillus acidophilus SLAM AK001, were introduced into the FIMM system with gut microbiota from a beagle model. Findings highlight the system's capacity to mirror and modulate the gut environment, evidenced by an increase in beneficial bacteria like Lactobacillus and Faecalibacterium and a decrease in the pathogen Clostridium. The study also verified the system's ability to facilitate accurate interactions between probiotics and commensal bacteria, demonstrated by the production of short-chain fatty acids and bacterial metabolites, including amino acids and gamma-aminobutyric acid precursors. Thus, the results advocate for FIMM as an in vitro system that authentically simulates the intestinal environment, presenting a viable alternative for examining gut microbiota and metabolites in companion animals.

• Journal of Food Hygiene and Safety
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• v.39 no.4
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• pp.312-321
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• 2024

Tomato is a widely distributed, cultivated, and commercialized vegetable crop. Recently, an increasing trend has been observed in the consumption of transgenic crops with enhanced functional components. However, consumer concerns regarding genotoxicity have been increasing. This study examined the genotoxicity of transgenic tomato (LTT) using the CRISPR/Cas9 system through a bacterial reverse mutation assay, chromosomal aberration assay, and mammalian micronucleus test. In the bacterial reverse mutation assay, LTT did not induce mutagenicity in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537, or Escherichia coli WP2uvrA, irrespective of the presence or absence of S9. LTT did not cause clastogenic or aneugenic chromosomal abnormalities during metaphase in CHL cells. Moreover, LTT did not increase the frequency of micronucleated polychromatic erythrocytes in the polychromatic erythrocytes. These findings can be used as a foundation to assess the genotoxicity of transgenic crops using the CRISPR/Cas9 system in the future.

• Applied Biological Chemistry
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• v.11
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• pp.1-27
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• 1969

More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.602-609
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• 2015

In this study, for modernization of Korean traditional fermented milk, Tarak was made using four kinds of commercial Makgeolli based on the ancient cookbook Suwoonjabbang. Samples of Tarak were periodically collected during 24 h of fermentation at $37^{\circ}C$. After fermentation, changes in pH, titration acidity, and viscosity were analyzed. Fermentation metabolites, including organic acids and free sugars, were analyzed by HPLC. Numbers of yeast and lactic acid bacteria during 24 h of fermentation were measured. The pH of Tarak significantly decreased (P<0.01), whereas its acidity significantly increased (P<0.01) during fermentation. The viscosity increased during 8~24 h of fermentation until curd was separated in Tarak. The level of ethanol increased from 0.37~0.52 mg/mL to 0.51~0.71 mg/mL during 24 h of fermentation. Lactic acid and lactose were the major organic acid and free sugar in Tarak, respectively. The number of lactic acid bacteria increased from 5.23~6.25 log CFU/mL to 9.87~10.41 log CFU/mL at the beginning during 24 h of fermentation. The number of yeast increased from 5.14~6.47 log CFU/mL to 6.99~7.73 at the beginning during 24 h of fermentation at $37^{\circ}C$. The major strains of Tarak were Pediococcus acidilactici, Lactobacillus fermentun, Lactobacillus curvatus, and Saccharomyces cerevisiae. Therefore, we concluded that Tarak was a fermented milk by both lactic acid bacteria and yeast, which was similar to koumiss or kefir.

• Clinical and Experimental Pediatrics
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• v.48 no.3
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• pp.260-265
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• 2005

Purpose : During epidemics, influenza induces a high mortality and morbidity, and when influenza is prevalent, it is revealed by increased pneumonia, hospitalization due to influenza-like illness, and mortality in community. We aimed at the isolation of influenza virus and prevalence period in Busan from 2000 to 2002. Methods : For 3 years from 2000 to 2002, we analyzed the patterns of influenza virus, the occurrence distribution of influenza by age and sex and the prevalence period after cultivating the examined materials from throat smears and snivel, collected from patients in St. Benedict Hospital Pediatrics Department, from 10 monitoring hospitals, and from 16 public health centers. Results : For three years, a total of 209 strains of influenza virus were isolated. In 2000, there were A/sydney/05/97(H3N2)-like, A/Beijing/262/95(H1N1)-like and B/Harbin/07/94-like. In 2001, there were A/Panama/2007/99(H3N2)-like and A/Newcaledonia/20/99(H1N1)-like. In 2002, there were A/Panama/2007/99(H3N2)-like, A/Newcaledonia/20/99(H1N1)-like, B/Beijing/243/97, B/Honkong/22/2001 and B/Sichuam/379/99. The occurrence distribution by sexes were 14 males and 25 females in 2000, 23 males and 33 females in 2001, 57 males and 57 females in 2002. As for the occurrence distribution by ages, 0-10 years made up 48.4 percent in 2000, 11-20 years 33.93 percent in 2001, and below 10 years was 64.91 percent in 2002. As for the occurrence distribution by month, the rate was once high in January and somewhat high in April and by June, when there happened to be various viruses, though there was a low rate in 2000. On the other hand, the virus was concentrated in February and March in 2001. And in 2002, it happened high twice, in March and November. Conclusion : Influenza virus revealed frequent antigenic changes and infect children, especially those below 10 years of age from late fall to early spring. So we should consider appropriate prevention in children.

• Korean Journal of Environmental Agriculture
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• v.27 no.2
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• pp.156-162
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• 2008

Salmonellosis is a major bacterial zoonosis that causes a variety of disease syndromes, self-limited enteritis to fatal infection in animals and food-borne infection and typhoid fever in humans. Recently, the emergence of multidrug resistant strains of Salmonella spp. causes more serious problems in environment and public health. The present study was investigated the antibacterial effect of Houttuynia cordata ethanol extract(HCEE) for murine salmonellosis. In the cytotoxic effect of HCEE on RAW 264.7 cells, there was no detectable effect with any concentrations between 25 and 100 ${\mu}g/ml$ after 8 h incubation. The bacteriocidal effect of HCEE was not showed on a Salmonella enterica serovar Typhimurium(S. typhimurium). HCEE makes morphological change of the RAW 264.7 cells, and there was significant decreased bacterial uptake and intracellular replication within Salmonella infected cells. And further nitric oxide(NO) production of Salmonella infected RAW 264.7 cells with HCEE was decreased comparing to RAW 264.7 cells without HCEE until 8 h post infection. Oral administration of HCEE showed a therapeutic effect for S. typhimurium infected BALB/c mice. The mortality of HCEE treated mouse was 80% until 12 days, while that of HCEE untreated mouse was 100 % until 8 days after lethal dose of S. typhimurium infection. These data suggested that HCEE has a potency treatment for intracellular replicative pathogen including salmonellosis, brucellosis, tuberculosis, listeriosis etc., and the application of HCEE makes new strategies for safety medicine development without antibiotic resistance bacterial appearance and residue problem in food and solves the public health problem from antibiotic mis- and over use.

• The Korean Journal of Physiology
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• v.23 no.2
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• pp.253-261
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• 1989

Responses of atrial natriuretic peptide (ANP), aldosterone and renin release to acute volume expansion were compared in normotensive Wistar and spontaneously hypertensive rat (SHR) fed low or high-sodium diet (2 or 25 mmol Na/100 g diet). Experimental diets were fed for 6 weeks from 7-week-old and the growth rate was similar in all groups. In the morning of the experiment, catheters were inserted under ether anesthesia in femoral artery for pressure recording and blood collection, femoral vein for saline infusion, and bladder for urine collection. Then, the rats were placed in restraining cages. When the rats were recovered from anesthesia and the arterial pressure became stabilized, control urine and blood samples were collected. Then, 0.9% saline was infused for 30 min for volume expansion (3% BW). Arterial pressure was significantly higher in the high-sodium SHR but there was no difference between the two groups of Wistar rats. Control plasma levels of Na, K, ANP, renin activity, and hematocrit were not different among the 4 groups. However, plasma aldosterone level was significantly higher in the low-sodium groups. Wistar low-sodium rats showed approximately two times higher plasma aldosterone level than the SHR counterpart. Volume expansion produced a marked increase in plasma ANP level, especially in the high-sodium groups. The low-sodium groups of both strains showed approximately two-fold increase in plasma ANP level. Following a volume expansion plasma aldosterone level and renin activity decreased in all groups. There was a significant logarithmic positive correlation between plasma renin activity and aldosterone concentration. The low-sodium rats produced a greater increase in aldosterone release by small increase in plasma renin than did the high-sodium rats. The low- and high-sodium rats produced a similar degree of diuresis and natriuresis after volume expansion. However, SHR produced a greater natriuresis than did the Wistar rats. The above results indicate that regulatory mechanisms of ANP, aldosterone and renin release are different between the normotensive and hypertensive rats, and between the low- and high-sodium groups.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.3
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• pp.325-331
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• 2012

The dual inoculation response of soybean with rhizobium and mycorrhiza was examined in pot vermiculite and field soils. In order to select a symbiotically compatible mycorrhiza with Bradyrhizobium japonicum, a highly germinating spore among 60 strains from 32 upland soils in southern part of Korea was obtained in Acaulospora sp., Gigaspora sp. and Glomus sp., respectively. As a result of dual inoculation of Glycin max cv. Dajangkong and Eunhakong both with $1{\times}10^8$cells of B. japonicum YCK 213 and 10 spores of each mycorrhiza in vermiculite pot, only Glomus sp. treatment together with the rhizobium showed significant increase ($P{\leqq}0.05$) both in shoot dry wt and nodule mass of not Eunhakong but Dajangkong. In red-yellow soils with pH 5.2($1:5H_2O$) and 203 mg of Lancaster P per kg of soil, in which $10^3$ cells of B. japonicum and $10{\pm}0.2$ spores of mycorrhizae per gram of soil were naturalized, grain yield of G. max cv. Dajangkong was increased to 3.9% by dual inoculation both of $4.8{\times}10^6$cells of B. japonicum and 10 spores of mycorrhizae per two seeds under condition applied with 30 kg $P_2O_5$ and 34 kg $K_2O$ per hectare compared to conventionally fertilized plot (2.75 MT $ha^{-1}$) added with 30 kg N $ha^{-1}$. However, there was not significant.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.172-178
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• 2013

The strains were added to the feed in the concentration of $10^3$, $10^5$, and $10^7$ CFU/kg and 2% of fishes were given the feed twice a day (8 AM and 5 PM) for 12 weeks. In result of the nonspecific immune response study to examine Respiratory burst activity, Lysozyme activity and Phagocytosis activity every two weeks until the end of the study, all test samples showed greater activities than control samples and improved immune activity with Bacillus sp. IS-2. The mortality test performed by artificial infection using Streptococcus iniae, a pathogenic bacterium, after the completion of this study also showed over 55% greater survival rate in all test samples. In result of performing PCR using the universal primer to verify that the probiotic stays in the intestines of the fishes, all test samples showed PCR product of 1,465 bp. Based on the above findings, it was concluded that Bacillus sp. IS-2 in the feed improved farmed flatfish's immune system and resistance against diseases as the probiotics. Also, the physiological indicators discovered by this study would be useful for identifying the mechanisms of probiotics.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.256-262
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• 2015

In order to study the effect of the receptor protein (SeaR), which is isolated from Saccharopolyspora erythraea, we introduced the SeaR gene to Streptomyces virginiae as host strains. An effective transformation procedure for S. virginiae was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP, and $ermEp^{\ast}$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. virginiae by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Transformants of S. virginiae containing the SeaR gene were confirmed by PCR and transcriptional expression of the SeaR gene in the transformants was analyzed by RT-PCR, respectively. And, we examined the production time of virginiamycins in the culture media of both the transformants and the wild type. The production time of virginiamycins in the wild type and transformants was the same. When 100 ng/ml of synthetic $VB-C_6$ was added to the state of 6 or 8 hour cultivation of wild type and transformants, respectively, the virginiamycins production was induced, meaning that the virginiamycins production in the wild type was detected 2 h early than transformants. From these results, SeaR expression was also affected to virginiamycins production in transformants derived from S. virginiae. In this study, we showed that the SeaR protein worked as a repressor in transformants.