• Title/Summary/Keyword: Agar dilution method

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Studies on the Antifungal Antibiotics Produced by a Streptomyces sp. (Part 4) The Occurrence of Tetraene Substance and Its Physiological Properties (Streptomyces sp. 가 생산하는 항진균성 항생물질에 관한 연구 (제4보) Tetraene계 항진균성 항생물질의 생성및 그의 성장)

  • Ko, Young-Hee;Bae, Moo
    • Microbiology and Biotechnology Letters
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    • v.10 no.3
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    • pp.211-215
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    • 1982
  • Streptomyces griseorubiginosus var. soyoensis previously identified, produced two kinds of antifungal antibiotics, trans-cinnamamide and another new substance. The latter was identified to be a new substance of tetraene family by establishment of UV, IR, NMR, mass spectra and chemical reactions and rotatively named as Tetraene KM-A. Through an antimicrobial activity test using serial agar dilution method, Tetraene KM-A showed strong growth inhibitory activity against fungi and yeasts, but not against procaryotes tested. The inhibitory action of Tetraene KM-A on fungi was remarkably ineffective when some of sterols were added to the cultural media. $LD_{50}$ of the Tetrene KM-A to mice and rats by intravenous injection were 84.3 and 90.4 mg/kg respectively. $LD_{50}$ to mice by oral feeding was 1503mg/kg.

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Distribution of Pseudomonas-Derived Cephalosporinase and Metallo-β-Lactamases in Carbapenem-Resistant Pseudomonas aeruginosa Isolates from Korea

  • Cho, Hye Hyun;Kwon, Gye Cheol;Kim, Semi;Koo, Sun Hoe
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1154-1162
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    • 2015
  • The emergence of carbapenem resistance among Pseudomonas aeruginosa is an increasing problem in many parts of the world. In particular, metallo-$\beta$-lactamases (MBLs) and AmpC $\beta$lactamases are responsible for high-level resistance to carbapenem and cephalosporin. We studied the diversity and frequency of $\beta$-lactamases and characterized chromosomal AmpC $\beta$lactamase from carbapenem-resistant P. aeruginosa isolates. Sixty-one carbapenem-resistant P. aeruginosa isolates were collected from patients in a tertiary hospital in Daejeon, Korea, from January 2011 to June 2014. Minimum inhibitory concentrations (MICs) of four antimicrobial agents were determined using the agar-dilution method. Polymerase chain reaction and sequencing were used to identify the various $\beta$-lactamase genes, class 1 integrons, and chromosomally encoded and plasmid-mediated ampC genes. In addition, the epidemiological relationship was investigated by multilocus sequence typing. Among 61 carbapenem-resistant P. aeruginosa isolates, 25 isolates (41.0%) were MBL producers. Additionally, 30 isolates producing PDC (Pseudomonas-derived cephalosporinase)-2 were highly resistant to ceftazidime (MIC50 = $256{\mu}g/ml$) and cefepime (MIC50 = $256{\mu}g/ml$). Of all the PDC variants, 25 isolates harboring MBL genes showed high levels of cephalosporin and carbapenem resistance, whereas 36 isolates that did not harbor MBL genes revealed relatively low-level resistance (ceftazidime, p < 0.001; cefepime, p < 0.001; imipenem, p = 0.003; meropenem, p < 0.001). The coexistence of MBLs and AmpC $\beta$-lactamases suggests that these may be important contributing factors for cephalosporin and carbapenem resistance. Therefore, efficient detection and intervention to control drug resistance are necessary to prevent the emergence of P. aeruginosa possessing this combination of $\beta$-lactamases.

Studies on Salmonella Isolated from Cattle (우(牛) 유래(由來)의 Salmonella속균(屬菌)에 대하여)

  • Jung, Suk-chan;Choi, Won-pil
    • Korean Journal of Veterinary Research
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    • v.26 no.1
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    • pp.79-85
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    • 1986
  • This paper deals with the isolation of Salmonella on 3 herds during the period from May 1984 to May 1985. Isolated Salmonella were examined for serotypes, adhesive fimbriae, antibiotic susceptibility and detection of R plasmid. The results were as follows; Of total 1505 cattle, 24 Salmonella were isolated from 18 cattle (1.2%) and their serotypes were S. enteritidis (9 strains), S. derby(4 strains), S. infantis (1 strain), $C_1$ group(8 strains), $C_2$ group(1 strain) and untypable(1 strain). Twelve strains of Salmonella were isolated from 227 cattle with diarrhea and their serotypes were S. enteritidis(4 strains), S. derby(3 strains), S. infantis(1 strain), $C_1$ group(3 strains) and untypable(1 strain). The isolation rate of Salmonella in cattle varied from 1.6 to 0.9% in 3 herds, it was higher in summer and autumn and lower with the age. Of total 24 strains, 23 were adhesive type 1 fimbriae. Antibiotic susceptibility test of Salmonella isolated was performed by the agar dilution method, using 9 antibiotics as follows: ampicillin(Am), chloramphenicol(Cm), gentamicin(Gm), kanamycin(Km), nalidixic acid(Na), rifampicin(Rf), streptomycin(Sm), sulfadimethoxine(Su) and tetracycline(Tc). All the strains tested were susceptible to Am, Cm, Gm, Km, Na, Rf and Tc. Of total 24 strains, 23(95.8%) were resistant to Su and 14(58.3%) to Sm. Of the 23 resistant Salmonella strains, all the strains were found to carry R plasmid. Among them, two strains which had the R plasmid conferring SmSu resistance was transferable at $25^{\circ}C$ only.

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Antimicrobial resistance and distribution of tetracycline resistance determinants in Escherichia coli isolated from aquatic birds (수생조류에서 분리한 대장균의 항균제 내성 및 Tetracycline 내성인자의 분포)

  • Cho, Jae-Keun;Lee, Sang-Min;Kim, Ki-Seuk
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.295-303
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    • 2008
  • One hundred and sixty nine Escherichia (E.) coli strains isolated from fecal samples of aquatic birds in Geumho river basin and Dalseong park were tested by agar dilution method to determine their susceptibility patterns to 14 antimicrobial agents. The distribution of tetracycline resistance determinants (tetA, tetB, tetC, tetD and tetE) were also examined by PCR in 76 tetracycline-resistant ($TC^r$) E. coli isolates. The high resistance was observed in tetracycline, cephalothin and ampicillin (45.0~36.7%). Resistance of E. coli isolates derived from Dalseong park to tetracycline, cephalothin, ampicillin and streptomycin (65.7~44.8%) were significantly higher than those isolated from Geumho river basin (31.4~14.7%). About seventy percent (70.4%) of the strains isolated were resistant to one or more drugs tested. Thirty (39.5%) of 76 $TC^r$ E. coli isolates which were resistant to one or more drugs transferred all or a part of their resistance patterns to the recipient strain of E.coli J53 by conjugation. All of $TC^r$ E. coli isolates contained at least one or more of 5 tet genes examined. The most common genes found in these isolates were tetA (60.6%) and followed by tetB (7.9%) and tetC (1.3%). However, tetD and tetE were not found in any of the isolates tested. Twenty one (27.6%) of $TC^r$ E. coli isolates had two determinants, tetA/tetB (20 strains), tetA/tetC (1 strain). And two strains (2.6%) contained three determinants (tetA/tetB/tetC).

Establishment of a linear regression equation for quantification of beta-hemolytic Escherichia coli in different media and survival of hemolytic Escherichia coli after blending with three different media

  • Kim, Jae Cheol;Pluske, John R.;Yoo, Jaehong;Heo, Jung Min
    • Korean Journal of Agricultural Science
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    • v.41 no.2
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    • pp.135-139
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    • 2014
  • Pathogenic E. coli associated post-weaning diarrhea (PWD) and edema disease are common diseases in commercially-housed weanling pigs. An enterotoxigenic E. coli (ETEC) oral challenge model has been used to mimic the physiological responses observed in commercial conditions. However, an oral challenge procedure has two major limitations: (1) the ETEC cell density is unknown at the point of oral inoculation, and (2) blending ETEC with traditional TSB (trypticase soy broth) is not palatable and hence decreases acceptability by piglets. Therefore, the purposes of this study were to (1) establish a regression equation that can be used for estimation of ETEC concentration in dilution media using the spectrophotometric measurement of cell density; and (2) examine survival of ETEC after blending either with TSB, sweetener or dextrose. A strain of ETEC (serogroup beta-hemolytic E. coli O149; K91; F4; toxins LT, STa, STb) was grown in TSB for 3.5 hours, centrifuged, the supernatant was discarded, and the ETEC pellet was then blended either with TSB (100 mL), sweetener (60 mL TSB + 40 mL fruit flavored concentrate), or dextrose (50 mL TSB + 50 mL dextrose; 0.5g/mL dextrose). Cell density was measured using the colorimetric method and also plated on a 5% sheep blood agar for counting of ETEC colony forming units at 0, 5, 35, 65 and 125 min after blending. The optical density at 600 nm explained 83% of ETEC colony forming units, indicating that the established linear equation (y= 6E+08x - 4E+07, P<0.004) can be used for robust quantification of ETEC cell density in TSB, sweetener and dextrose media. When ETEC was blended with sweetener and dextrose, survival of ETEC was decreased by 45% and 72% within 5 min post-blending. Therefore, further research is required to find out the suitable medium that has potential to improve palatability without compromising survival of ETEC.

In Vitro Anti-Helicobacter pylori Activity of Ethanol Extract of Sohamhyoongtang and Coptidis Rhizoma Total Alkaloids (소함흉탕 에탄올 추출물 및 황련 알칼로이드의 헬리코박터 파이로리에 대한 항균활성)

  • Lee, BaWool;Choi, MyungSook;Yim, DongSool;Choi, SungSook
    • Korean Journal of Pharmacognosy
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    • v.45 no.2
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    • pp.168-173
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    • 2014
  • The aim of this study was to evaluate the anti-helicobacter activity of the ethanol extract of Sohamhyoongtang (Coptidis Rhizoma, Pinelliae Tuber and Trichosanthis Semen) and Coptidis Rhizoma total alkaloids, which is one of the components of Sohamhyoongtang. Crude ethanol extract of Sohamhyoongtang (ESHHT) and Coptidis Rhizoma total alkaloids (CRTA) were used for this experiment. Five different types of H. pylori (including H. pylori 26695) were used as test strain. To determine anti-helicobacter activity, minimum inhibitory concentration (MIC) was determined by agar dilution method. The effect of ESHHT and CRTA on the gene expression of H. pylori was investigated by quantitative realtime-PCR (qRT-PCR). MICs of ESHHT against five H. pylori strains were $250{\sim}500{\mu}g/ml$ and MICs of CRTA against five H. pylori strains were $50{\sim}200{\mu}g/ml$. Four representative virulence genes of H. pylori, cagA, ureA, ureB and ureI were tested as target genes for qRT-PCR. According to the qRT-PCR results, both ESHHT and CRTA markedly repressed the expression of cagA gene of H. pylori 26695 (6.91 and 20 folds respecively). These results showed that the ESHHT and CRTA demonstrated antihelicobacter properties, suggesting their potential use in gastritis or duodenal ulcer.

Sensitivity of Pathogens of Bovine Udder Origin to Antibiotics (젖소 유방원(乳房源) 병원세균(病原細菌)의 항생물질(抗生物質)에 대(對)한 감수성(感受性))

  • Chung, Jong Sig;Cho, Sung Whan;Cho, Yoong Jun;Park, Cheong Kyu
    • Korean Journal of Veterinary Research
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    • v.19 no.1
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    • pp.75-83
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    • 1979
  • The sensitivities of 270 pathogens (124 Streptococci, 118 Staphylococci, 10 Corynebacterium pyogenes and 18 Escherichia coli) isolated from clinical or subclinical cases of bovine mastitis during lactation to 11 antibiotics were determined by the agar plate dilution method. All cultures of Streptococci were inhibited at $1.56{\mu}g/ml$ of ampicillin and 1.56 units/ml of penicillin G. Most of the cultures were inhibited at $3.125{\mu}g/ml$ of leukomycin, but were resistant to kanamycin, streptomycin, chloramphenicol, erythromycin and colistin at concentration of $12.5{\mu}g/ml$. More than 93% of the Staphlococcal cultures were sensitive to kanamycin, leukomycin, ampicillin, tetracycline, ledermycin and minocycline at concentrations of $12.5{\mu}g/ml$ or less, and sensitive to penicillin at concentration of 3.125 units/ml, but for more than 71% of the cultures to chloramphenicol, erythromycin and colistin the concentrations required to inhibit growth were 100 ${\mu}g/ml$ or higher. All 10 cultures of Corynebacterium pyogenes were inhibited by leucomycin, ampicillin and minocycline at concentration of $1.56{\mu}g/ml$ and by penicillin G at concentration of 0.78 units/ml, but all the cultures required at least $400{\mu}g/ml$ or higher of streptomycin, erythromycin and colistin for inhibition. More than 83% of E. coli cultures were sensitive to erythromycin and minocycline at concentration of $12.5{\mu}g/ml$, but resistant to leucomycin and chloramphenicol at concentration of $100{\mu}g/ml$.

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Antimicrobial Susceptibility of Escherichia coli Isolated from Fish Farms on the Southern Coast of Korea (남해안 어류양식장에서 분리된 Escherichia coli에 대한 항균제 감수성)

  • Son, Kwang-Tae;Oh, Eun-Gyoung;Park, Kun-Ba-Wui;Kwon, Ji-Young;Lee, Hee-Jung;Lee, Tae-Seek;Kim, Ji-Hoe
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.4
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    • pp.322-328
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    • 2009
  • Three-hundred and sixteen Escherichia coli strains from seawater, and a variety of farmed fishes, including oliver flounder (Paralichthys olivaceus), black rock fish (Sebastes schlegeli), red sea bream (Pagrus major) and sea bass (Lateolabrax japonicus) between May to October in 2004, were tested by agar dilution method to determine their susceptibility patterns to 17 antimicrobial agents. Overall, 92.1% of Escherichia coli isolates from samples showed antimicrobial resistance to at least one antimicrobial agent and the multiple resistance was seen in 173 isolates (54.7%). The resistance of E. coli isolates to tetracycline (74.1%) was highest, followed by cephalothin (69.9%), doxycycline (66.5%), streptomycin (47.2%), ampicillin (46.2%), cefazolin (31.6%), enrofloxacin (31.0%). norfloxacin (28.2%). The most frequent resistance pattern was TE-D-CF-CIP-ENO-NOR-AM-S-C-SXT-AmC-CZ (14.7%), followed by CF (6.2%), TE (5.1%), TE-CF (4.5%) in 177 isolates from fishes and TE-D-CF (7.2%) followed by TE-D-CF-S (5.8%), CF and TE-D-S (3.6%) in 139 isolates from seawater.

A Rapid Detection of Methicillin-Resistant Staphylococci by Polymerase Chain Reaction (Polymerase Chain Reaction을 이용한 Methicillin-resistant Staphylococci의 신속 검출)

  • 박진숙;박영진
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.306-311
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    • 2002
  • PCR of the mecA gene for the rapid detection of methicillin-resistant staphylococci was perfomed and compared with the antibiotic sensitivity test. A total of 43 strains of staphylococi from clinical specimens were used in this study. An antibiotic sensitivity test by the agar dilution method of NCCLS (The National Commitee for Clinical Laboratory Standard) was performed for the strains. Among them, 39 isolates were methicillin-resistant (MRS), and 4 isolates were methicillin-susceptible (MSS). With the exception for one strain (Staphylococcus cohnii, HRC2-4), all MRS strains amplified the expected 533 bp fragments of the mecA gene by PCR, However, one strain (Staphylococcus aureus, HSA1-10) that was classified as a sensitive strain by the antibiotic sensitivity test was mecA positive by PCR. All 35 methicillin-resistant Staphylococcus aureus (MRSA) strains were mecA positive, but overall, concordance between the results of the mecA PCR and antibiotic sensitivity test was 95.6%.

Antifungal activity of N-[1-(benzotriazol-1-yl)aryl]arylamine derivatives and quntitative structure-activity relationships(QSAR) (N-[1-(benzotriazol-1-yl)aryl]arylamine 유도체의 항균성과 정량적 구조활성 관계(QSAR))

  • Sung, Nack-Do;Kim, Kyoung-Hoon;Choi, Woo-Young;Kim, Hong-Ki
    • Applied Biological Chemistry
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    • v.35 no.1
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    • pp.14-22
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    • 1992
  • A series of new N-[1-(benzotriazol-1-yl)aryl]arylamine derivatives were synthesized and their antifungal activities $(pI_{50})$ in vitro against Pyricularia oryzae, Fusarium oxysporum f. sp. sesami, Valsa ceratosperma and Botrytis cinerea were dertermined by the agar medium dilution method. From the results of the quantitative structure-activity relationships $(QSAR_S)$ analysis, $hydrophobicity({\pi})$, $electronic({\Sigma\sigma})$ and molar $refractivity({\Sigma}M_R)$ parameter of X & Y-substituents on the phenyl group were also shown to be important factor in determining the variation in the antifungal activity. 4-Bromo group substituents (1d & 2b) were the most effective compounds and the $half-life(T_{1/2})$ on the hydrolysis of X(1) at netural pH was about 1.5 day. Molecular orbital(MO) functions of substrate compound, linear free energy relationships$(LFER_S)$ on the antifungal reactivity arid the results of molecular design were also discussed.

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