• Laboraroty Animal Research
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• v.34 no.4
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• pp.279-287
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• 2018

Placenta specific 8 (PLAC8, also known as ONZIN) is a multi-functional protein that is highly expressed in the intestine, lung, spleen, and innate immune cells, and is involved in various diseases, including cancers, obesity, and innate immune deficiency. Here, we generated a Plac8 knockout mouse using the CRISPR/Cas9 system. The Cas9 mRNA and two single guide RNAs targeting a region near the translation start codon at Plac8 exon 2 were microinjected into mouse zygotes. This successfully eliminated the conventional translation start site, as confirmed by Sanger sequencing and PCR genotyping analysis. Unlike the previous Plac8 deficient models displaying increased adipose tissue and body weights, our male Plac8 knockout mice showed rather lower body weight than sex-matched littermate controls, though the only difference between these two mouse models is genetic context. Differently from the previously constructed embryonic stem cell-derived Plac8 knockout mouse that contains a neomycin resistance cassette, this knockout mouse model is free from a negative selection marker or other external insertions, which will be useful in future studies aimed at elucidating the multi-functional and physiological roles of PLAC8 in various diseases, without interference from exogenous foreign DNA.

• Laboraroty Animal Research
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• v.34 no.4
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• pp.257-263
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• 2018

Trefoil factor 1 (TFF1, also known as pS2) is strongly expressed in the gastrointestinal mucosa and plays a critical role in the differentiation of gastric glands. Since approximately 50% of all human gastric cancers are associated with decreased TFF1 expression, it is considered a tumor suppressor gene. Tff1 deficiency in mice results in histological changes in the antral and pyloric gastric mucosa, with severe hyperplasia and dysplasia of epithelial cells, resulting in the development of antropyloric adenoma. Here, we generated Tff1-knockout (KO) mice, without a neomycin resistant ($Neo^R$) cassette, using the clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRSIPR/Cas9) system. Though our Tff1-KO mice showed phenotypes very similar to the previous embryonic stem (ES)-cell-based KO mice, they differed from the previous reports in that a reduction in body weight was observed in males. These results demonstrate that these newly established Tff1-KO mice are useful tools for investigating genetic and environmental factors influencing gastric cancer, without the effects of artificial gene insertion. Furthermore, these findings suggest a novel hypothesis that Tff1 expression influences gender differences.

• Animal Bioscience
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• v.35 no.1
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• pp.138-146
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• 2022

Objective: The objective of the current study was to investigate the influences of conventional (CO) and deep litter (DE) systems on antimicrobial resistance in fecal Escherichia coli (E. coli). Methods: A cross-sectional study was carried out to detect antimicrobial resistance to E. coli in swine fecal samples in CO and DE systems located in western and northeastern Thailand. Individual rectal swab samples were taken only from healthy pigs. A total of 215 individual and healthy pigs were randomly selected for isolation and antimicrobial susceptibility test of E. coli by the disc diffusion method. The test panel included amoxicillin (AMX), colistin, doxycycline (DOX), enrofloxacin, gentamicin (GEN), kanamycin, neomycin (NEO), and trimethoprim-sulfamethoxazole (SXT). Results: There were significant (p<0.05) lower resistance levels for GEN, NEO, and SXT in the DE farms compared to those in the CO farms. There was a lower number of antimicrobial resistance agents (p<0.001) in the DE farms compared to those in the CO farms. This result was consistent with those in western (p<0.01) and northeastern (p<0.01) Thailand. Overall, antibiograms of AMX-SXT and AMX-DOX-SXT were found in the CO (19.09% and 20.91%, respectively) and the DE (16.19% and 24.76%, respectively) farms. No antimicrobial resistance (5.71%) was found and AMX (13.33%) resistant pigs in the DE farms, whereas the pattern of AMX-GEN-SXT (6.36%) and AMX-DOX-GEN-SXT (11.82%) resistant pigs was found in the CO farms. Conclusion: The DE system for pig farming was superior to conventional pig farming by lowering the resistance level of fecal E. coli to GEN, NEO, and SXT, with decreasing the number of antimicrobial resistance agents and inducing a small proportion of pigs to be free from antimicrobial resistance.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.265-273
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• 2023

Background: The intestinal microbiota is an important regulator of bone health. In previous studies we have shown that intestinal microbiota dysbiosis, induced by treatment with broad spectrum antibiotics (ABX) followed by natural repopulation, results in gut barrier dysfunction and bone loss. We have also shown that treatment with probiotics or a gut barrier enhancer can inhibit dysbiosis-induced bone loss. The overall goal of this project was to test the effect of Korean Red Ginseng (KRG) extract on bone and gut health using antibiotics (ABX) dysbiosis-induced bone loss model in mice. Methods: Adult male mice (Balb/C, 12-week old) were administered broad spectrum antibiotics (ampicillin and neomycin) for 2 weeks followed by 4 weeks of natural repopulation. During this 4-week period, mice were treated with vehicle (water) or KRG extract. Other controls included mice that did not receive either antibiotics or KRG extract and mice that received only KRG extract. At the end of the experiments, we assessed various parameters to assess bone, microbiota and in vivo intestinal permeability. Results: Consistent with our previous results, post-ABX- dysbiosis led to significant bone loss. Importantly, this was associated with a decrease in gut microbiota alpha diversity and an increase in intestinal permeability. All these effects including bone loss were prevented by KRG extract treatment. Furthermore, our studies identified multiple genera including Lactobacillus and rc4-4 as well as Alistipes finegoldii to be potentially linked to the effect of KRG extract on gut-bone axis. Conclusion: Together, our results demonstrate that KRG extract regulates the gut-bone axis and is effective at preventing dysbiosis-induced bone loss in mice.

• Microbiology and Biotechnology Letters
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• v.50 no.4
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• pp.522-532
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• 2022

Cheonggukjang is a traditional fermented food in Korea, which is known to exert beneficial effects on health. In this study, we evaluated the effects of cheonggukjang fermented by Bacillus subtilis SCGB 574 (B574) on high fat diet (HFD)-deteriorated large intestinal health. Rats were fed with HFD or HFD supplemented with 10.1% cheonggukjang (B574). Fecal microbiota was analyzed based on 16S rRNA gene sequences, and the fecal and serum metabolome were measured using GC-MS. Our results showed that SCGB574 intake significantly reduced body weight, restored tight junction components, and ameliorated inflammatory cell infiltration. SCGB574 also shifted gut microbiota by increasing the abundance of short chain fatty acid producers such as Alistipes and Flintibacter, although it decreased the abundance of Lactobacillus. Serum and fecal metabolome analyses showed significantly different metabolic profiles between the groups. The top five metabolites increased by SCGB574 were i) arginine biosynthesis, ii) alanine, aspartate, and glutamate metabolism; iii) starch and sucrose metabolism; iv) neomycin, kanamycin, and gentamicin biosynthesis; and v) galactose metabolism. These results showed that cheonggukjang fermented by SCGB574 ameliorates adverse effects of HFD through improving intestinal health.

• Journal of Food Hygiene and Safety
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• v.13 no.3
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• pp.305-312
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• 1998

In order to investigate the species, serobiological characteristics and antibiotic sensitivity of Pseudomonas spp, we isolated Pseudomonas spp from 57 spring waters around Seoul area for spring, summer and autumn and identified Pseudomonas spp by biochemical characteristics and serological method. And also we tested the antibiotic sensitivity test by discdiffusion method. Of 57 spring waters tested, Pseudomonas spp were isolated from 33 spring waters(57.9%). Isolation rate of Pseudomonas spp in spring season was 28.1%, summer 21.1% and autumn 28.1%. Only 1 spring water was detected Pseudomonas spp in all seasons and 9 (15.8%) were detected for 2 seasons and 13 (22.8%) were for only 1 season. Isolation rate of Pseudomonas spp at Mt. Cheonggye was 50% and followed by Mt. Bookhan 35.7%, Mt. Daemo 33.3%, Mt. Dobong 29.6%, Mt. Surak 25.9%, Mt. Woomyun 22.2% and Mt. Bulam 7.4%. Of 44 Pseudomonas spp, 22 strains (50%) were identified by Ps. putida, Ps. aeruginosa, Ps. fluorescens and Ps. mendocina were identified 6 strains (13.6%), respectively. 4 strains (9.1%) were identified by Ps. aureofaciens. Of 6 Ps. aeruginosa, serotype A was 2 strains, B, E, G, and K was 1 strain, respectively. Of 44 Pseudomonas spp, resistance rate to amoxicillin was 90.9% and followed by chloramphenicol 84.1%, tetracycline 84.1%, carbenicillin 81.8%, nalidixic acid 68.2%, neomycin 38.6%, streptomycin 31.8%, gentamicin 4.6%, kanamycin 4.6% and colistin 2.3%. Ps. aeruginosa was more sensitive to carbenicillin than other Pseudomonas spp isolated from spring waters in Seoul area but more resistant to kanamycin, and Ps. aureofaciens was no resistant to streptomycin. Among multiple drug resistance, resistance to 5 drugs was 31.8%, 4 drugs 15.9%, 7 drugs 13.6%, 1 drug and 2 drugs 4.6%, and 8 drugs 2.3%, respectively. The multiple resistance patterns detected highestly were NA-CB-C-TE-AMC (18.2%), NA-CB-N-C-TE-AMC (13.6%), CBC-TE-AMC (11.4%) and NA-CB-N-C-TE-AMC-S (9.1%).

• The Journal of the Korean Society for Microbiology
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• v.9 no.1
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• pp.7-11
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• 1974

The authors identified fifty-eight Shigella cultures among 1644 cultures and specimens of enteric pathogens collected from all over the country in 1973. Fifty-one out of fifty-eight cultures belonged to Shigella flexneri and the rest to Shigella sonnei. None of cultures belonging to either subgroup A or C was detected in 1973. Of fifty-one cultures of Shigella flexneri twenty-six cultures were $B_{2a}$, which were isolated in Seoul area and Kwangwon-Do. The rest were $B_{3a}$ which were isolated in Jeonla-bug-Do and Kangwon-Do. It would not be possible to understand that there might not have been the cases or carriers of Shigella in the areas where the organisms were not isolated in 1973 and that there might not have been any other serotypes existing in the country, although there was a quite disparity found in the distribution between different areas and in the detection of the serotypes as shown in Table 1. Concerning the biochemical properties there were only two cultures showing positive arginine decarboxylase test among $B_{2a}$, and there were three cultures of trehalose negative cultures, one of rhamnose positive culture and one of glycerol positive culture observed, which were considered to be unusual. All the Shigella cultures were sensitive to nitrofurantoin, cephalosporin and ampicillin, and resistant to colistin, bacitracin and neomycin. Majority of them showed sensitive results to gentamycin, and the majority of Shigella $B_{3a}$ appeared to be sensitive to chloramphenicol, tetracycline, oxytetracycline and doxycycline, but the majority of $B_{2a}$ and Shigella sonnei were observed resistant to those antibiotics by means of the In-Vitro tests.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.1
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• pp.71-76
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• 1998

To know the antibiotic specificity of a Dinoflagellate, Cochlodinium polykrikoides, we investigated the survival time of C. polykrikoides against several concentrations of antibiotics and judged the selective specificity of antibiotics based on the $LT_50$ ($50\%$ of lethal time). The result showed that C. polykrikoides was sensitive to tetracycline and chloramphenicol, and resistant to polymixin-B, ampicillin, penicillin-G, dihydrostreptomycin, and neomycin. In the case of sensitive antibiotics to C. polykrikoides, tetracycline and chloramphenicol, the safety concentrations of both antibiotics were determined and the antibiotic specificity based or the plotted survival curve was analyzed. Before antibiotic treatment, we tested the antibiotic susceptibility of the contaminated bacterial population in tile culture of C. polykrikoides, and decided the proper kinds of antibiotics and concentrations before percoll-centrifugation. By percoll-centrifugation, we reduced bacteria, removed fungi, collected the algal pellet, and made axonic culture by antibiotic cascade procedure based on the result of antibiotic susceptibility test. We observed that axonic C. polykrikoides culture entered the logarthmic phase of growth when cell density was over 740 cells/ml and propagated to 5,800 cells/ml maximally. Divisions per day, k value of C. polykrikoides represented a good index for growth at the low density of cells. There was a highest k value shift before reaching to the logarithmic phase. We suggested that the preceeding highest k value shift stage is a good indicator for accurate broadcasting for red. tide blooming in the field, and the stage is also a good time for controlling red tide blooming in the filed, either.

• The Journal of the Korean Society for Microbiology
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• v.7 no.1
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• pp.9-15
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• 1972

The authors identified 45 Shigella cultures among 63 suspectable cultures obtained from the cases showing dysentery or the like symptoms in various parts of the country during the period from February to September, 1971. Of 45 cultures, 36 cultures belonged to Subgroup B, 7 cultures to Subgroup C and 2 cultures to Subgroup D. There was none of cultures belonging to Subgroup A in 1971, although the authors detected one culture of that among 41 identified Shigella in 1967. Of 36 cultures belonging to Subgroup B, 27 cultures were $B_{2a}$ and 3 cultures of each $B_{3c},\;B_{4a}$ and $B_y$ were serotyped, respectively. Of 7 cultures beloning to Subgroup C, one was $C_{11}$ and other six cultures were $C_{15}$ and all S. boydii were isolated in Seoul area. It might be possible to suggest that there was a tendency of decreasing prevalence of S. dysenteriae and of increasing prevalence of S. boydii in Korea, although the number of cultures. tested were very much limited to conclude, if it was compared with the results obtained by the authors in 1967 that the fourty-one identified Shigella were composed of one culture of S. dysenteriae, 36 cultures of S. flexneri, 4 cultures of S. sonnei and none of S. boydii. According to the results obtained from the biochemical tests, the positive ratio of Indol tests in Subgroup B was 31/36, which could be higher than 9/36 observed in 1961, but which was nearly correlated with the result published by Ewing and his colleagues in U.S.A. The positive rates of both sorbitol and raffinose were lower than that observed by Ewing and his colleagues in S. flexneri. Regarding with the sensitivity of Shigella cultures to the antibiotics being widely used in the country, 46.2% were sensitive to chloramphenicol, which was very much higher than that observed by Park, and 85.8% were sensitive to ampicillin, which would be the drug of choice according to the results from the In Vitro tests.

• Korean Journal of Food Science and Technology
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• v.14 no.4
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• pp.291-300
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• 1982

In order to develop effective and simple sanitation method for the extention of shelf-life of fresh poultry meat, the effect of sanitizers, sanitation methods and packaging materials on the extention of shelf-life of poultry meats was observed at the $4^{\circ}C$ and room temp$(10{\sim}20^{\circ}C)$. The results are summarized as follows: 1. The autochonous skin microflora of poultry, before processing, were believed to be removed or killed during the scalding and plucking, and exposed dermal tissue was contaminated by microorganisms from the subsequent stages of processing. 2. In the final stage of poultry processing, total viable counts of microorganisms and coliforms were averaged to $3.5{\times}10^4/cm^2$ and $400/cm^2$, respectively. 3. The refrigerated shelf-life of fresh whole poultry carcasses at $3\;to\;4^{\circ}C$ was extended to 7 to 16 days compared to control with the various treatments of some sanitizers by dipping freshly chilled carcasses for 5 min or spraying 1 liter of sanitizers per carcasses. In the case of storage at $10\;to\;15^{\circ}C$, the shelf-life of poultry carcasses was extended to one to two days by the sanitation treatments compared to control. 4. Spraying sanitation was more effective than dipping sanitation, and 5 minutes dipping and one liter spraying per carcass were enough for effective sanitation of poultry carcasses in most sanitizers. 5. The packaging with an oxygen impermeable polyvinylidene chloride extended the shelf-life to 10 days and 5 days with polyethylene compared to control. When poultry carcasses were sanitized by continuous spraying with one liter of 30 ppm of chlorine and another one liter of 5% of potassium sorbate, packaged with polyvinylidene chlorlde were extended to about 30 days compared to control.