• Title/Summary/Keyword: ${\beta}$-lactam

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Purification and Characterization of β-Lactamase Secreted from Bacillus sp. J105 Strain having β-Lectam Antibiotics Resistance. ((β-lactam계 항생물질 저항성을 지닌 Bacillus sp. J105 균주로부터 분비되는 베타 락탐 분해효소의 정제 및 특성)

  • Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Min-Jeong;Lee, Young-Choon;Lee, Jai-Heon;Joo, Woo-Hong;Choi, Yung-Hyun;Lim, Hak-Seob;Kim, Jeong-In;Seo, Kwon-Il;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.6
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    • pp.845-851
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    • 2008
  • ${\beta}-Lactamase$, secreted from Bacillus sp. J105 strain was purified to a single band on SDS-PAGE by ammonium sulfate precipitation, ion exchange column chromatography and gel-filtration. The molecular weight of the purified enzyme was 31 kDa on SDS-PAGE and its isoelectric point was 7.35. Optimal pH and temperature for enzymatic reaction were 5 and $40^{\circ}C$, respectively. As a result of total amino acid composition analysis of the purified enzyme, Gly and Ala were occupied 14.1 and 13.3 mole %, respectively. Km and Vmax value of purified enzyme were 1.33 mM and 0.36 mM/ml using ampicillin as a substrate, respectively.

PERIODONTOPATHIC BACTERIA AND ANTIBIOTIC RESISTANCE GENES OF ORAL BIOFILMS IN CHILDREN (어린이 치면세균막에서 치주질환원인균과 항생제 내성유전자의 출현율)

  • Kim, Seon-Mi;Choi, Nam-Ki;Cho, Seong-Hoon;Lee, Seok-Woo;Lim, Hoi-Jeong;Lim, Hoi-Soon;Kang, Mi-Sun;Oh, Jong-Suk
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.2
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    • pp.170-178
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    • 2011
  • The purpose of this study was to assess the prevalence of periodontopathic bacteria and resistance determinants from oral biofilm of children. Subgingival dental plaque was isolated from 87 healthy children, and PCR was performed to determine the presence of 5 periodontal pathogens including P. gingivalis, T. forsythia, T. denticola, F. nucleatum, A. actinomycetemcomitans, and nine resistance genes including tet(Q), tet(M), ermF, aacA-aphD, cfxA, $bla_{SHV}$, $bla_{TEM}$, vanA, mecA. 1. The prevalence of F. nucleatum, T. forsythia. and P. gingivalis was 95.4%, 55.2%, and 40.2%, respectively. In addition. the prevalence of A. actinomycetemc omitans was 5.7%, while T. denticola was 3.4%. 2. In analysis of antibiotic resistance determinants. cfxA, $bla_{TEM}$ and tet(M) were detected in all the samples tested. It was also found that the prevalence of tet(Q) showing tetracycline resistance. $bla_{SHV}$ associated with resistance to ${\beta}$-lactams, ermF exhibiting erythromycin resistance, and, vanA resulting vancomycin resistance was 88.5%, 29.9% 87.4%, and 48.5%, respectively. The aacA-aphD gene showing resistance to aminoglycosides and mecA gene harboring methicillin resistance exhibited the lowest prevalence with 9.2%. 3. In a correlation analysis between periodontopathic pathogens and antibiotic resistance determinants, it was found that there was a significant correlation between T. forsythia and $bla_{SHV}$. Also, P. gingivalis and vanA showed a correlation. Finally, tet(Q) and ermF showed a significant correlation (phi: 0.514) while mecA and vanA also showed a correlation(phi: 0.25).

Analysis of Initial Choice Antibiotics Efficacy in Diabetic Foot Infection (당뇨병성 족부 감염 환자의 초기 선택 항생제 효율성)

  • Lee, Doo-Hyung;Han, Seung-Hwan;Park, Min-Jung
    • Journal of Korean Foot and Ankle Society
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    • v.13 no.2
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    • pp.146-149
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    • 2009
  • Purpose: Foot infections are common complications in patients with diabetes. The patients are usually immune-compromised; therefore the pathogens could be resistant to narrow spectrum antibiotics. Those drugs, however, are categorized as specially managed antibiotics, and access are difficult without confirming of the pathogens. Our aim was to analyze the common pathogens in diabetic foot infection and figure out the proper antibiotics. Materials and Methods: We studied 68 patients treated with diabetic foot infection. The pathogens which caused the infection and their sensitivity to initial antibiotics were analyzed. We also investigated the change of the antibiotics after the confirming of the culture result and average time to get the result. Results: Among the 68 patients, 56 (82%) received cephalosporin and beta-lactam antibiotics. Only 12 (18%) who were confirmed the drug resistant pathogens from previous culture, were treated with broad spectrum antibiotics such as vancomycin and tazoperan. Average culture study time was 6 days. Methicillin-resistant staphylococcus aureus (MRSA) was cultured in 19 patients (28%), Methicillin-resistant coagulase negative staphylococcus (MRCNS) in 11 patietns (17%), pseudomonas in 11 patients (17%). Total 44 (65%) including 3 of other antibiotics resistant pathogen needed broad spectrum antibiotics. Thirty two patients (47%) were resistant to initial antibiotics.irt follow up culture, 2 MRSA and 2 MRCNS were found. The antibiotics resistant pathogens were confirmed in 48 (71%) patients at last. Conclusion: Diabetic patients with foot infection need proper antibiotics from initial treatment. The proper broad spectrum antibiotics should assigned to the patients from the first time without the confirming of the culture results.

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Studies on the Penicillinase Produced by a Streptomyces sp. (Part I). Optimal Conditions for the Penicillinase Production by Streptomyces sp. YS-40. (Streptomyces sp.가 생산하는 Penicillinase 에 관한 연구 (제1보) Streptomyces sp. YS-40에 의한 Penicillinase의 생산조건)

  • 도재호;김상달;이동의
    • Microbiology and Biotechnology Letters
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    • v.10 no.3
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    • pp.177-184
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    • 1982
  • Studies were carried out to investigate the optimal culture conditions for the production of penicillinase using a strain of Streptomyces sp. isolated from soil, YS-40. Among the carbon and nitrogen sources, glucose and L-asparagine increased the peniciilinase production. The addition of M $n^{++}$, $Ca^{++}$ and L $i^{+}$ increased the enzyme production, but depressed by F $e^{+++}$, F $e^{++}$, $Mg^{++}$, Z $n^{++}$, A $g^{++}$, $Ba^{++}$ and S $n^{++}$. L-Leucine slightly increased the enzyme production but L-histidine, L-methionine depressed. Among the vitamins riboflavine, i-inositol, hesperidine, niacin-amide, biotin, folic acid, DL-$\alpha$-lipoic acid increased the enzyme formation. The addition of cephradine, cephalexin, ampicillin, cloxacillin more increased the enzyme formation than that of other$\beta$-lactam antibiotics and antibiotics. Optimal pH and temperature on the enzyme formation was pH 7.0 and 28$^{\circ}C$ respectively Amount of the enzyme production reached at maximum with incubation for 3 days on the optimal condition.

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High Resolution Whole Genome Multilocus Sequence Typing (wgMLST) Schemes for Salmonella enterica Weltevreden Epidemiologic Investigations

  • Tadee, Pakpoom;Tadee, Phacharaporn;Hitchings, Matthew D.;Pascoe, Ben;Sheppard, Samuel K.;Patchanee, Prapas
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.162-170
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    • 2018
  • Non-typhoidal Salmonella is one of the main pathogens causing food-borne illness in humans, with up to 20% of cases resulting from consumption of pork products. Over the gastroenteritis signs, multidrug resistant Salmonella has arisen. In this study, pan-susceptible phenotypic strains of Salmonella enterica serotype Weltevreden recovered from pig production chain in Chiang Mai, Thailand during 2012-2014 were chosen for analysis. The aim of this study was to use whole genome sequencing (WGS) data with an emphasis on antimicrobial resistance gene investigation to assess their pathogenic potential and genetic diversity determination based on whole genome Multilocus Sequence Typing (wgMLST) to expand epidemiological knowledge and to provide additional guidance for disease control. Analyis using ResFinder 3.0 for WGS database tracing found that one of pan-susceptible phenotypic strain carried five classes of resistance genes: aminoglycoside, beta-lactam, phenicol, sulfonamide, and tetracycline associated genes. Twenty four and 36 loci differences were detected by core genome Multilocus Sequence Typing (cgMLST) and pan genome Multilocus Sequence Typing (pgMLST), respectively, in two matching strains (44/13 vs A543057 and A543056 vs 204/13) initially assigned by conventional MLST and Pulsed-field Gel Electrophoresis (PFGE). One hundread percent discriminant ability can be achieved using the wgMLST technique. WGS is currently the ultimate molecular technique for various in-depth studies. As the findings stated above, a new of "gold standard typing method era" for routine works in genome study is being set.

The neuroprotective mechanism of ampicillin in a mouse model of transient forebrain ischemia

  • Lee, Kyung-Eon;Cho, Kyung-Ok;Choi, Yun-Sik;Kim, Seong Yun
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.2
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    • pp.185-192
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    • 2016
  • Ampicillin, a ${\beta}$-lactam antibiotic, dose-dependently protects neurons against ischemic brain injury. The present study was performed to investigate the neuroprotective mechanism of ampicillin in a mouse model of transient global forebrain ischemia. Male C57BL/6 mice were anesthetized with halothane and subjected to bilateral common carotid artery occlusion for 40 min. Before transient forebrain ischemia, ampicillin (200 mg/kg, intraperitoneally [i.p.]) or penicillin G (6,000 U/kg or 20,000 U/kg, i.p.) was administered daily for 5 days. The pretreatment with ampicillin but not with penicillin G significantly attenuated neuronal damage in the hippocampal CA1 subfield. Mechanistically, the increased activity of matrix metalloproteinases (MMPs) following forebrain ischemia was also attenuated by ampicillin treatment. In addition, the ampicillin treatment reversed increased immunoreactivities to glial fibrillary acidic protein and isolectin B4, markers of astrocytes and microglia, respectively. Furthermore, the ampicillin treatment significantly increased the level of glutamate transporter-1, and dihydrokainic acid (DHK, 10 mg/kg, i.p.), an inhibitor of glutamate transporter-1 (GLT-1), reversed the neuroprotective effect of ampicillin. Taken together, these data indicate that ampicillin provides neuroprotection against ischemia-reperfusion brain injury, possibly through inducing the GLT-1 protein and inhibiting the activity of MMP in the mouse hippocampus.

Antibacterial Effect of Ipyo-san against Methicillin-Resistant Staphylococcus aureus (입효산(立效散)의 Methicillin-Resistant Staphylococcus aureus에 대한 항균활성에 관한 연구)

  • Yoon, Jae-Hong;Choi, Yeun-Ju;Jeong, Seung-Hyun;Shin, Gil-Cho
    • The Journal of Internal Korean Medicine
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    • v.34 no.3
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    • pp.278-288
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    • 2013
  • Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.

Identification and staphylococcal cassette chromosome mec (SCCmec) type of methicillin-resistant coagulase-negative staphylococci isolated from chickens (닭에서 분리된 methicillin-resistant coagulase-negative staphylococci의 동정 및 staphylococcal cassette chromosome mec (SCCmec) type)

  • Kong, Shin-Koog;Yook, Sim-Yong;Lee, Geon-Taek;Kim, So-Yeon;Hong, Young-Un;Jung, Yoon-Taek;Lee, Jung-Hwa;Kim, Hee-Jeong;Hwang, Soo-Myung;Chang, Kyung-Soo
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.233-240
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    • 2010
  • Methicillin-resistant coagulase-negative staphylococci (MRCNS) were isolated from the respiratory sites of chickens in 4 farms and slaughter house located in Chungnam provinces. Isolation of coagulase-negative staphylococci (CNS) was positive for 61 (26.6%) of the 229 chickens tested, and isolation of MRCNS was positive for 17 (27.9%) of the isolated CNS. A total of 17 MRCNS isolates were selected and subjected to identification. Of the 17 MRCNS isolates selected, 6 were identified as Staphylococcus cohnii, 2 as S. saprophyticus, 3 as S. simulans, 3 as S. lentus, 2 as S. carnosus, and 1 as S. xylosus. The MRCNS isolates were resistant to many beta-lactam antibiotics, and some isolates were also resistant to macrolide and aminoglycoside antibiotics. The mecA gene was detected in some isolates of each MRCNS strains. The mecA-positive isolates were classified into five staphylococcal cassette chromosome mec (SCCmec). SCCmec types I to IV were detected in isolates from chickens.

A Case of Group A Streptococcal Pneumonia with Empyema and Pericardial Effusion (폐농양과 심막삼출이 동반된 A군 연구균에 의한 폐렴 1례)

  • Chun, Yoon Hong;Lee, Soo Yong;Choi, Sang Lim;Jeong, Dae Chul;Chung, Seung Yeon;Kang, Jin Han
    • Pediatric Infection and Vaccine
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    • v.11 no.2
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    • pp.202-207
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    • 2004
  • Group A streptococcus, also known as Streptococcus pyogenes, is a common bacterial pathogens of the upper respiratory tract and skin infections in children, but this organism is a less common cause of pneumonia, pericarditis. However, pneumonia that is caused by Streptococcus pyogenes, may be rapidly progressive course with developing severe consequences. It may be focal but often is bilateral and diffuse involvement of lung. Empyema is commonly developed, and pleurocentesis often yields thin, watery fluid that continues to flow out when a chest tube is inserted. Antimicrobial resistance to the ${\beta}$-lactam antibiotics has not been reported against group A streptococci, whereas increasing resistance to the macrolides seems to be directly related to the consumption of specific antimicrobial agent use in the community. Clindamycin resistance is uncommon but does occur. We experienced one case of group A streptoccoccal pneumonia with empyema and pericardial effusion, and treated successfully with amoxicillin-clavulanate, clindamycin and roxithromycin.

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Detection of Methicillin-Resistant Staphylococcus aureus by In Vitro Enzymatic Amplification of MecA and FemA Gene (메티실린 내성 황색 포도상 구균에서 mecA, femA 유전자의 임상적 의의)

  • Park, Jung-Eun;Kim, Taek-Sun;Park, Su-Sung;Kim, Eun-Ryoung;Kim, Il-Su;Ann, Il-Young;Kim, Young-Jin;Kim, Jae-Jong;Kang, Sung-Ok;Park, Han-Ho
    • Pediatric Infection and Vaccine
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    • v.3 no.2
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    • pp.133-138
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    • 1996
  • Purpose : In the treatment of MRSA infection, rapid detection of MRSA is extremely important. The mecA gene codes the new drug resistant polypeptides called PBP2' which mediates the clinically relevant resistance to all beta-lactam antibiotics. The identical mecA gene has been found in coagulase-negative staphylococcus with the methicillin-resistant phenotype. On the other hand, the femA gene was absent from coagulase negative staphylococcus strains with the methicillin resistant phenotype. This study is aimed at early detection and definite diagnosis of MRSA. Methods : A total of 24 MRSA strains were studied. All strains were tested for antimicrobial susceptibility and purified DNA. We amplified both mecA and femA genes by PCR in 24 strains. Results : In MRSA all the 16 strains (100%) carried femA gene and 11 strains (68.7%) carried mecA gene. In contrast, in methicillin sensitive staphylococcus all the 8 strains (100%) carried femA and only 3 strains (37.5%) were detected mecA. Conclusions : As results, there are difference in the phenotype and genotype of methicillin resistance by PCR of mecA and femA. Such disparities between methicillin resistance and the presence of mecA gene suggest the presence of control gene of the mecA.

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