To investigate the population dynamics and survival of Genus Vibrio, population densities of aerobic saprophytic bacteria and Vibrio groups were measured 4 times in the intertidal waters of the Yellow Sea near Kunsan from November, 1997 to June, 1998. The distribution of heterotrophic bacteria during the survey periods by plate count and direct count method ranged from 1.2$\pm$0.6$\times$10$^3$~2.0$\pm$1.5$\times$10$^4$CFU ml$^1$and from 6.0$\pm$4.0$\times$10$^{5}$ ~1.9$\pm$1.5$\times$10$^{7}$ cells ml$^1$, respectively. Vibrio groups were distributed in the range of 1$\times$10 and 6$\pm$2.2$\times$10$^2$CFU ml$^1$. The proportion of Vibrio groups to total heterotrophic bacteria was between 0.1 and 6% during the survey periods. A total of 51 isolates was obtained from TCBS agar plates and identified to species level by Biolog Identification System$^{TM}$. As a result, dominant genera were V, mediterranei, V aitguillarum, tr metschnikovii, and V. parahaemolyticus, and isolates were clustered into 26 groups based on the relatedness of average linkage clustering method at 70% level. As for the susceptibility of 51 isolates to 7 kinds of antibacterial agents (gentamicin, ampicillin, chlorarnphenicol, streptomycin, kanamycin, tetracycline, carbenicillin), 96% of isolates showed high resistance to more than one antibiotics and 65% of isolates contained a plasmid, of which size was observed greater than 12 kb, The number of cells of 3 tested strains (V. anguillarum, V. vulnificus, and V. metschnikovii) in filtered aged seawater decreased by approximately 1 to 5 orders of magnitude during 30-d incubation. In most cases, the numbers of cells decreased rapidly until day 3, then decreased slowly by day 30. The number of cells incubated at 15$^{\circ}C$ showed higher survival than those at 4$^{\circ}C$ and $25^{\circ}C$. These results may be considered for the basic supporting data in the risk assessment of vibriosis in summer.r.
The Study was conducted to develope the low temperature tolerant methane-producing bacteria(LTTB) and to increase the efficiency of anaerobic fermentation for the treatment of agricultural and livestock wastes at low temperature. The samples were collected from muddy soil, water logged sediment, organic layer and anaerobic sludge at three latitudes, $34.8{\sim}37.4\;^{\circ}N(Korea)$, $41.4\;^{\circ}N(USA)$ and $54.5{\sim}56.9\;^{\circ}N(Canada)$. They were used for determination of the methanogenesis rates for isolation and identification of the LTTB. The methanogenesis rate of smaples at low temperature were higher in the cellulose medium than methanol medium. The methanogenesis rate in the samples of subarctic region were $15{\sim}19$ moles/ml during 30 days at low temperature($8\;^{\circ}C$), whereas not detected in the samples of temperate region. The methanogenesis rate in the enrichment culture of subarctic samples were inhibited by the $40\;{\mu}g/ml$ of streptomycin + vancomycin or ampicillin + oleandomycin which were not effect to the methanogens. An inhabitation of high temperature tolerant methane producing bacteria was identified in the samples of temperate region, whereas that of the LTTB growing at $8{\sim}13^{\circ}C$ was identified in the subarctic region.
The Journal of the Korean Society for Microbiology
/
v.17
no.1
/
pp.21-34
/
1982
One hundred and twenty-one strains each of Escherichia coli isolated from stools of 60 patients who received various antimicrobial drugs in hospital for more than one week and apparently healthy 60 students who have no history of taking antimicrobial drugs during recent one month, were tested for their resistance to 13 antimicrobial drugs. The frequency of resistance strains was highest to tetracycline with 69.2%, and followed by streptomycin(Sm), sulfisomidine(Su), chloramphenicol(Cm), ampicillin(Ap), and carbenicillin(Cb) in the decreasing order, ranging from 61.2% to 39.3%. Strains resistant to kanamycin(Km), cephaloridine(Cr), and trimethoprim(Tp) occupied about one-fourth of strains, and only four strains were resistant either one or more of nalidixic acid, gentamicin and amikacin, and no strain was resistant to rifampicin. The frequency of resistant strains to Cm, Ap, Km, Cr, and Cb was much higher among patient isolates than student strains, but strains resistant to the other drugs showed almost the same frequencies between patient and student isolates. There was a marked difference in average minimum inhibitory concentrations of between resistant and susceptible strains, suggesting that the resistance to drugs is the plasmid origin. Seventy-six percent of strains were resistant to one to 10 drugs tested, and no much difference was observed between strains from patients and students. However, strains resistant to four or more drugs were much more frequently found among patient isolates than student strains, with the increasing tendency of multiply resistant strains among patient isolates following the increase in the number of resistant drugs. The transfer of drug resistance by conjugation was tested and 98 strains(67.5%) among 145 which were resistant to two or more drugs were found to transfer their drug resistance to E. coli. Among 74 strains resistant to 7 or more drugs, all except one transferred the resistance, and the number of strains with transferable resistance decreased, as the number of resistant drugs decrease. A R plasmid from randomly selected p13 strain was tested for the incompatibility group, and the plasmid was classified into Inc F II. R plasmM DNA bands were identified by polyacrylamide gel electrophoresis.
Thirty five Salmonella enterica serovar Typhimurium strains were isolated from diarrheic patients and pigs in Gyeongbuk area from 2003 to 2004. All 35 strains (17 strains from diarrheic patients and 18 from pigs) were resistant to more than one drug and most of strains isolated from pigs were resistant to ampicillin, ohloram-phenicol, streptomycin, sulfamethoxazole-trimethoprime, tetracyclin and nalidixic acid. Each isolate was also screened or the presence of class I, II and III integron gene cassettes. Among 35 strains,3 out of 17 strains isolated from diarrheic patients, carried dhfrX-orfF-aadA2 integron gene cassette and among 18 strains isolated from diseased pigs, 11 strains carried dhfrX-orfF-aadA2 integron gene cassette and 1 strain carried aadA2 integron only. But any class II and class II integron gene cassette were not detected in 35 strains. Thirty five strains were divided by five pulsotypes. Thirty one strains out of thirty five were pulsotype A. Among the remaining 4 strains, one each strain belonged to pulsotype B, C, D and pulsotype E. This data of pulsotypes showed that the widespread of pulsotype A, Salmonella enterica serovar Typhimurium in human and pigs in Gyeongbuk area may have been caused by the dissemination of a few epidemic strains in this area. Thirteen strains contain dhfrX-orfF-aadA2 integron gene cassette showed pulsotype A and one strain contains dhfrX-orfF-aadA2 integron gene cassette showed pulsotype B. One strain contains aadA2 integron showed pulsotype E. But fifteen strains do not contain any integron showed pulsotype A.
Koh, Won-Jung;Kwon, O Jung;Kang, Eun Hae;Suh, Gee Young;Chung, Man Pyo;Kim, Hojoong;Chung, Myung Jin;Kim, Tae Sung;Lee, Kyung Soo;Lee, Nam Yong;Park, Young Kil;Bai, Gill Han
Tuberculosis and Respiratory Diseases
/
v.57
no.3
/
pp.234-241
/
2004
Background : There has been a gradual increase in the number of newly diagnosed cases of Mycobacterium avium complex (MAC) pulmonary disease. However, the optimal therapeutic regimen for the disease has not yet established and there is no report about the treatment outcome of MAC pulmonary disease in Korea. This study examined the effect of clarithromycin-based regimen in patients with pulmonary MAC disease without a HIV infection. Materials and Methods : Fifty-six patients with pulmonary MAC disease were diagnosed according to the American Thoracic Society criteria from January 2000 to December 2003 at this hospital. Of these patients, 15 were treated with clarithromycin, rifampin, and ethambutol for more than 6 months, together with streptomycin initially (first 6 months) in 8 patients. Results : Six months after the treatment, the sputum cultures converted from positive to negative in 8 patients (53%) and the radiological findings improved in 10 (67%). At 12 months 4 patients (44%) achieved sputum negative conversion and 6 patients out of 9 patients (67%) who were treated for more than 12 months showed radiological improvement. Overall, the sputum findings converted to negative in nine patients (60%) who underwent medical treatment. A pulmonary resection was successfully performed in one patient. Only one patient discontinued the treatment due to side effects such as gastrointestinal intolerance and optic neuritis. Conclusion : A combined regimen containing clarithromycin is relatively safe and tolerable even in the elderly outpatients. However, the results of this combined chemotherapy were unsatisfactory and new companion drugs for MAC pulmonary disease are needed. A resection may be considered for localized disease.
Mallory, Larry M.;Yuk, Chang-Soo;Liang, Li-Nuo;Alexander, Martin
Korean Journal of Soil Science and Fertilizer
/
v.16
no.4
/
pp.358-367
/
1983
Antibiotic-resistant strains of Salmonella typhimurium and Klebsiella pneumoniae died readily after their addition to raw sewage, but they grew in sterilized sewage. The decline was not a result of antibiotic stresses, and because the bacteria were able to survive in large numbers for at least 15 days in solutions containing no organic nutrients, it was not a result of competition. Toxin production, bacteriophages, and Bdellovibrio did not cause the disappearance of the two bacterial species. A decline was also evident if the sewage was first passed through a $3-{\mu}m$ filter or treated with cycloheximide or cycloheximide plus nystatin, but protozoa developed under these conditions. Little or no decline occurred if the sewage was filtered and treated with the eucaryotic inhibitors before adding S. typhimurium or K. pneumoniae, and protozoa were not detected. S. typhimurium increased in abundance if cycloheximide, streptomycin, and erythromycin or large amounts of glucose were added to sewage. Tetrahymena thermophilus did not significantly reduce the population of S. typhimurium in buffer when the density of the bacterium was about $10^4/ml$. However, when more than $10^8$ Enterobacter agglomerans cells per ml were added to the buffer, T. thermophilus reduced the abundance of E. agglomerans and S. typhimurium to $10^6$ and 10/ml, respectively. The density of S. typhimurium was further decreased by a second increment of E. agglomerans cells. The disappearance of S. typhimurium and K. pneumoniae from sewage thus is the result of predation by protozoa. It is proposed that predators will eliminate a prey species from a natural environment when an alternate prey is present at concentrations above the threshold number for active feeing by the predator and when the rate of growth of the prey is less than the rate of predation.
Kim, Yong-Nam;Yang, Kyu-Ho;Oh, Jong-Suk;Chung, Jin
Journal of the korean academy of Pediatric Dentistry
/
v.27
no.2
/
pp.361-369
/
2000
Enterococcus is a normal inhabitant of the human oral cavity, the vagina, and the gastrointestinal tract. Four isolates of Enterococcus in this study were identified as E. durans. These bacteria were characterized and the interaction of these bacteria with the important oral bacteria as like S. mutans and S. oralis was studied as follows. 1. The carbohydrate fermentation test and biochemical test showed similar results in 4 isolates. 2. The susceptibility test against erythromycin, penicillin, tobramycin, ampicillin, teicoplanin, ciprofloxacin, vancomycin, gentamicin, kanamycin, and streptomycin showed to be susceptible in all four isolates. 3. The optical density of absorbance at 550 nm was 1.405 in the culture of S. mutans in disposable cuvette, whereas being 0.855, 0.867, 0.797, and 1.083 in the combined culture of S. mutans and each E. durans. 4. The mean weight of produced artificial plaque on the wires in the beaker was $1566{\pm}103mg$ in culture of S. mutans only, whereas being reduced to $44{\pm}5mg,\;41{\pm}12mg,\;34{\pm}7mg,\;and\;38{\pm}12mg$ in the combined culture of S. mutans and each E. durans. The viable cells were $2.0\times10^9$ per ml in the culture of S. mutans wheras being $2.0\times10^7\;to\;6.0\times10^7$ per ml in the combined culture S. mutans and E. durans. 5. The viable cells were $2.1\times10^8$ per ml in the culture of S. oralis, wheras being $1.4\times10^7\;to\;7.0\times10^7$ per ml in the combined culture of S. oralis and E. durans. 6. Plasmid of about 60 kb was isolated in three isolates of E. durans. These results suggested that E. durans isolated from the oral cavity inhibited the replication of S. mutans and formation of artificial plaque, while inhibiting the replication of S. oralis a little.
The Journal of the Korean Society for Microbiology
/
v.14
no.1
/
pp.27-37
/
1979
Ninety-five strains of Shigella, 70 of Salmonella paratyphi A, and 230 of Salmonella typhi were tested for their resistance to drugs. Also studied was the inhibition and elimination of drug resistance. All except one strain of Shigella consisted of 79 Sh. flexneri and 16 Sh. sonnei were multiply resistant to chloramphenicol, tetracycline, streptomycin, and splfisomidine. Among them, 70 strains were resistant to ampicillin and carbenicillin, 80 to trimethoprim-sulfamethoxazole, 22 to nalidixic acid, and one to kanamycin, but strain resistant to gentamicin, cephaloridine, and rifampin was not encountered. All strains of S. paratyphi A and S. typhi were susceptible to drugs tested, except sulfisomidine and rifampin, for which all S. paratyphi A were slightly resistant to sulfisomidine and the majority of S. paratyphi A and S. typhi were slightly resistant to rifampin. Approximately 80% of multiply drug-resistant Shigella transferred their resistance to E. coli by conjugation, and the resistance was considered to be mediated by R plasmids. The frequency of transfer of drug resistance varied by donor strains and recipients, but not by selecting drugs. Resistance to nalidixic acid was not transferred by conjugation to the recipients. Drug-resistant Shigella strains successively subcultured in nutrient agar stabs contained clones resistant to drugs and those susceptible to drugs, but the ratio of resistant and susceptible clones varied by strains. The multiply drug-resistant S. typhi and Shigella strains were found to not lose completely their drug resistance by subculture in media. Acriflavine has some effect on the elimination of drug resistance mediated by R plasmids, but the effect varied markedly by strains. Atabrine has no effect among strains tested. The combination of drugs increased the drug actions in majority of cases with synergistic or additive effects.
The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.
Yoo Min Ho;Jeong Joon Beom;Kim Eun Heui;Lee Hyoung Ho;Jeong Hun Do
Korean Journal of Fisheries and Aquatic Sciences
/
v.35
no.2
/
pp.115-121
/
2002
To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.
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