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Lipopolysaccharide-Deficient Mutants of <i>Salmonella enterica</i>Have Increased Sensitivity to Catechins

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Advances in Microbiology 2013年第5期3卷 399-402页

作者： Miho Yoshii Akira Okamoto michio ohta Department of Bacteriology Nagoya University Graduate School of Medicine Nagoya Japan

Antimicrobial activity is one of the well-known biological characteristics of catechins, the main extract of green tea leaves. It is thought that catechins intercalate into the bacterial cell membrane and damage the lipid bilayer. However, the association between catechins and lipopolysaccharides, which consist of an O side chain, core oligosaccharide, and lipid A, has not been previously investigated. In this study, we evaluated the catechin sensitivity of Salmonella enterica mutants that lack the O side chain and have core oligosaccharides of different lengths. These rough mutants were more sensitive to catechins than a bacterial strain with intact lipopolysaccharide. We conclude that the O side chain and core oligosaccharide play an important role in protecting Gram-negative bacteria against the antimicrobial activity of catechins.

关键词： Antimicrobial Activity Catechins Lipopolysaccharide Gram-Negative Bacteria

The Two-Component Sensor Protein CovS Affects Penicilling Susceptibility by Modulation of Cell-Wall Synthesis in Streptococcus pyogenes

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Journal of Biosciences and Medicines 2015年第3期3卷 50-55页

作者： Masaaki Minami Syun Syun Torii michio ohta Department of Bacteriology Graduate School of Medical Sciences Nagoya City University Nagoya Japan School of Nursing Sugiyama Jyogakuen University Nagoya Japan

In Streptococcus pyogenes, we have described the two-component signal transduction sensor and regulatory systems, CovR/S affect the antimicrobial susceptibility including penicillin G before. But the mechanism how two-component sensor protein CovS modulates penicillin G susceptibility has not been elucidated till date. This study aimed to determine how the CovS affected penicillin G susceptibility in Streptococcus pyogenes by northern blot analysis. At first, we investigated the covS mRNA expression under penicillin G induction. We found that the decrease of covS mRNA expression under Penicillin G stimulation. Next we investigated the expression of cell wall synthesis gene, pbp2a and glmM with use of covS knockout mutants from emm1 Streptococcus pyogenes strain 1529. We found that the cell-wall synthesis gene expression of the ?covS mutant strain were lower than that of the wild-type strain. Furthermore the expression of glmM mRNA gene was lower than the expression of pbp2a mRNA gene in the ?covS mutant strain. The covS-complemented strain almost restored the mRNA expression compared to covS mutant strain. The two-component sensor protein CovS affects the susceptibility to penicillin G in Streptococcus pyogenes by modulation of cell-wall synthesis.

关键词： Streptococcus Pyogenes CovS pbp2a glmM

Comparative Study of Urosepsis-Associated Escherichia coli in Tertiary Care University Hospital in the Central Region of Japan from 2008 to 2011

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Journal of Biosciences and Medicines 2016年第6期4卷 18-23页

作者： Masaaki Minami Naoki Wakiyama Minoru Ohhashi Yukio Wakimoto michio ohta Department of Bacteriology Graduate School of Medical Sciences Nagoya City University Nagoya Japan Department of Clinical Investigation Nagoya City University Hospital Nagoya Japan School of Nursing Sugiyama Jyogakuen University Nagoya Japan

Escherichia coli infection is important cause of morbidity and mortality. Urosepsis is most commonly caused by Escherichia coli. It is generally reported to have low mortality rates and favorable outcomes compared with sepsis induced in other organ/ system or tissues. This study was conducted to find out the clinical characteristics of urosepsis-associated Escherichia coli isolates at tertiary care university hospital in the central region of Japan from 2008 to 2011. Escherichia coli was identified by standard laboratory procedure. Antimicrobial susceptibility testing was performed by micro dilution assay according to CLSI recommendation. We analyzed the relationship between medical records appended to clinical species and bacterial data by using a statistical method. Of one hundred five Escherichia coli, fifty-three were from urosepsis and fifty-two were from other disease. The ratio of male to female from urosepsis tended to be lower than that from others. Forty-two isolates were from outpatient and sixty-three were from inpatient. The ratio of inpatient to outpatient from urosepsis was significant lower than that from others. With respect to age, the numbers of elder patients were higher than those of others. Most of the Escherichia coli isolates were from the emergency room followed by urology, and general medicine. The ratio of urosepsis to no urosepsis at urology department was significant higher than that at other departments. With respect to antimicrobial susceptible pattern, the ratio of urosepsis to no urosepsis about only aztreonam resistant was significant higher than that about other antibiotics. We need enough attention to be paid to urosepsis, especially female and elder patients.

关键词： Escherichia coli Urosepsis Antimicrobial Resistance Susceptibility Epidemiology

Trend of Invasive Streptococcus agalactiae at Tertiary Care Hospital in Japan

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Journal of Biosciences and Medicines 2016年第3期4卷 15-20页

作者： Masaaki Minami Hideki Nishiyama Shinobu Ikegami Takuya Hattori Makoto Kawashima michio ohta Department of Bacteriology Graduate School of Medical Sciences Nagoya City University Nagoya Japan Department of Clinical Laboratory Japanese Red Cross Nagoya Daiichi Hospital Nagoya Japan Department of Clinical Laboratory Japanese Red Cross Nagoya Daini Hospital Nagoya Japan School of Nursing Sugiyama Jyogakuen University Nagoya Japan

Streptococcus agalactiae causes various severe infectious diseases such as sepsis, meningitis, and streptococcal toxic shock-like syndrome. Though Streptococcus agalactiae infection has been increasing recently, the comprehensive characteristic investigation of invasive Streptococcus agalactiae isolated in tertiary care hospitals has not been nearly performed in Japan. In this study, we investigated the clinical characteristics and antimicrobial susceptible patterns of 88 Streptococcus agalactiae isolated at two tertiary care hospitals during 2009-2015 in Japan. There was no significant differences between genders in our study. Two-third Streptococcus agalactiae were isolated from over age 60. Total mortality rate was 19% and invasive Streptococcus agalactiae-associated death cases have occurred every year after 2011. All Streptococcus agalactiae were completely susceptible toampicillin. Total non-susceptible rates of erythromycin, minocycline, levofloxacin and trimethoprim-sulfamethoxazole in this study were approximately 30%, 44%, 37%, and 7%, respectively. Our results suggest the need for continuous antimicrobial susceptibility survey of Streptococcus agalactiae.

关键词： Streptococcus agalactiae Susceptibility Antimicrobial Resistance Epidemiology

Clinical Characteristics of Invasive Methicillin-Resistant Staphylococcus aureus at General Hospital in the Central Region of Japan from July 2014 to June 2015

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Journal of Biosciences and Medicines 2015年第9期3卷 9-14页

作者： Masaaki Minami Ryoko Sakakibara Taichi Imura Hideo Morita Naoto Kanemaki michio ohta Department of Bacteriology Graduate School of Medical Sciences Nagoya City University Nagoya Japan Department of Clinical Investigation Daido Hospital Nagoya Japan Department of Gastroenterology Daido Hospital Nagoya Japan School of Nursing Sugiyama Jyogakuen University Nagoya Japan

Methicillin-resistant Staphylococcus aureus is crucial pathogen caused severe invasive infection disease. This study was conducted to find out the prevalence and antimicrobial susceptibility pattern of invasive Methicillin-resistant Staphylococcus aureus isolates at general hospital in the central region of Japan from July 2014 to June 2015. Methicillin-resistant Staphylococcus aureus was identified by standard laboratory procedure. Antimicrobial susceptibility testing was performed by micro dilution assay according to CLSI recommendation. Invasive Methicillin-resistant Staphylococcus aureus disease was defined as isolation of bacteria from a normally sterile body site. One hundred seventy-one methicillin-resistant Staphylococcus aureus were isolated among which 95 (55.6%) were from inpatient and 76 (44.4%) were from outpatient. The age incidence of (0 - 1) years, (1 - 10) years, (11 - 40) years, (41 - 60) years and >60 years age groups were 18 (10.5%), 41 (24.0%), 15 (8.8%), 5 (2.9%), and 92 (53.8%) respectively. There was significant difference of age distribution between invasive and noninvasive disease in 0 - 1 years group and 11 - 40 years age group. Positive samples were received mostly from the pediatrics (56/32.7%), respiratory medicine (25/14.6%) and general medicine (25/14.6%). We also found the significant differences of department between invasive and noninvasive disease in pediatrics, dermatology, and surgery. Arbekacin, teicoplanin, and vancomycin were the most active antibiotics with 100% susceptible rates in our study. Our study revealed that erythromycin and gentamicin were more antimicrobial effective in invasive disease than in noninvasive disease significantly. Methicillin-resistant Staphylococcus aureus infection spreads worldwide easily and inadequate use of antibiotics contributes to uptake of their new antimicrobial resistance. Continuous antimicrobial surveys are need for guiding policy on the adequate use of antibiotics to reduce the morbid

关键词： Methicillin-Resistant Staphylococcus aureus Susceptibility Antimicrobial Resistance Invasive Disease

Clinical Characteristics of Vaginal Discharge Associated Streptococcus pyogenes at General Japanese Hospital

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Journal of Biosciences and Medicines 2018年第1期6卷 9-16页

作者： Masaaki Minami Ryoko Sakakibara Mika Watanabe Hideo Morita Naoto Kanemaki michio ohta Department of Bacteriology Graduate School of Medical Sciences Nagoya City University Nagoya Japan Department of Clinical Investigation Daido Hospital Nagoya Japan Department of Gastroenterology Daido Hospital Nagoya Japan School of Nursing Sugiyama Jyogakuen University Nagoya Japan

Streptococcus pyogenes (S. pyogenes) is popular pathogen caused various infectious disease. This study was conducted to find out the recent clinical characteristics of S. pyogenes isolates from vaginal discharge at Japanese hospital. S. pyogenes was identified by standard laboratory procedure. Antimicrobial susceptibility testing was performed by microdilution assay according to CLSI recommendation. Total 61 S. pyogenes were isolated from vaginal discharge. The major age incidence among 0 - 10 years age group was 14 (23%), among 21 - 30 years age group, 7 (11.5%), among 31 - 40 years age group, 13 (21.3%), among 51 - 60 years age group, 8 (13.1%), and among 61 - 70 years age group, 11 (18%). The numbers of ciprofloxacin and clarithromycin no-susceptible S. pyogenes from 0 - 10, 31 - 40, and 61-70 years age group were higher than those from other groups. The numbers of clindamycin and minocycline no susceptible S. pyogenes from 0 - 10, and 21 - 40 years age group were higher than those from other group. Furthermore, our study revealed that clarithromycin was completely ineffective in 21 - 30 years age group (p S. pyogenes because the trend of infectious disease epidemiology is always changing drastically.

关键词： Streptococcus pyogenes Susceptibility Antimicrobial Resistance Vaginal Discharge Clarithromycin

LicT Modulates Biofilm Formation of Streptococcus pyogenes

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Journal of Biosciences and Medicines 2014年第9期2卷 1-7页

作者： Masaaki Minami Hiroshi Takase Ryoko Sakakibara Taichi Imura Hideo Morita michio ohta Department of Bacteriology Graduate School of Medical Sciences Nagoya City University Nagoya Japan Core Laboratory Graduate School of Medical Sciences Nagoya City University Nagoya Japan Department of Clinical Investigation Daido Hospital Nagoya Japan Department of Gastroenterology Daido Hospital Nagoya Japan School of Nursing Sugiyama Jyogakuen University Nagoya Japan

Streptococcus pyogenes frequently causes purulent infections in humans. Biofilm formation is an important virulence property of S. pyogenes because of decreased susceptibility of bacteria to antibiotic treatment. Biofilm is composed of various types of matrix including glycocalyx which is an important exocellular matrix material related to bacterial sugar metabolism. A putative antiterminator protein, LicT (Spy0571), is one of the components of the glucose-independent β-gluco-side-specific phosphotransferase system (PTS). Although the PTS, a carbohydrate metabolic system, may play a role in biofilm formation, the relationship between LicT and biofilm formation has not yet been elucidated. Here, we evaluated whether LicT affected biofilm formation in modified chemically defined medium (CDMM) supplemented with glucose or β-glucoside:salicin. We created licT- and licT-complemented mutant strains from S. pyogenes 1529. Although the licT mutant strain tended to have higher growth rate than wild-typestrain in CDMM with glucose, it had a significant lower growth rate than the wild-type strain in CDMM with salicin. In addition, the licT mutant exhibited lower biofilm formation in CDMM containing salicin than the wild-type strain by 96 well plate analysis and confocal laser scanning microscopic analysis. Our results suggest that LicT plays an important role in biofilm formation of S. pyogenes.

关键词： Streptococcus pyogenes LicT Biofilm

Prevalence and Antimicrobial Susceptibility Pattern of Streptococcus pneumoniae at General Hospital in the Central Region of Japan from December 2013 to February 2014

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Journal of Biosciences and Medicines 2014年第6期2卷 12-17页

Streptococcus pneumonia infection is important cause of morbidity and mortality. This study was conducted to find out the prevalence and antimicrobial susceptibility pattern of Streptococcus pneumoniae isolates at general hospitalin the central region of Japan from December 2013 to February 2014. Streptococcus pneumoniae was identified by standard laboratory procedure. Antimicrobial susceptibility testing was performed by micro dilution assay according to CLSI recommendation. One hundred fifty-three Streptococcus pneumoniae were isolated among which 80 (52.2%) were males and 73 (47.8%) were females. Nasal discharge (134%/87.6%) contributed more than other biological materials. The age incidence of (0 - 1) years, (1 - 10) years, (11 - 40) years, (41 - 60) years and >60 years age groups were 26 (17.0%), 110 (71.9%), 3 (2.0%), 10 (6.5%), and 4 (2.6%) respectively. Positive samples were received mostly from the pediatrics (137%/89.5%), respiratory medicine (12%/7.8%) and lowest from gastroenterology (1%/0.6%) and neurology (1/ 0.6%) department. Vancomycin and rifampicin were the most active antibiotics with 100% susceptibility rates. The next best were levofloxacin, penicillin G and ceftriaxone. Our study revealed that 82 Streptococcus pneumonia isolates had multidrug resistant ability (53.6%). Streptococcus pneumoniae infection spreads among community easily and inappropriate use of antibiotics contributes to their resistance. Continuous antimicrobial susceptible surveys are essential to guide policy on the adequate use of antibiotics to reduce the morbidity and mortality and reduce the emergency of antimicrobial resistance.

关键词： Streptococcus pneumoniae Susceptibility Antimicrobial Resistance Multi Drug Resistance

Analysis of Pyelonephritis-Associated Beta Hemolytic Streptococcus in Japan

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Journal of Biosciences and Medicines 2018年第12期6卷 45-52页

作者： Masaaki Minami Hideki Nishiyama Shinoba Ikegami Takuya Hattori Makoto Kawashima michio ohta Department of Bacteriology Graduate School of Medical Sciences Nagoya City University Nagoya Japan Department of Clinical Laboratory Japanese Red Cross Nagoya Daiichi Hospital Nagoya Japan Department of Clinical Laboratory Japanese Red Cross Nagoya Daini Hospital Nagoya Japan School of Nursing Sugiyama Jyogakuen University Nagoya Japan

Pyelonephritis is an inflammation of the kidney tissue, calyces, and renal pelvis. It is commonly caused by bacterial infection that has spread up the urinary tract. Common bacteria are Escherichia coli and Enterococcus faecalis. Beta hemolytic streptococci also cause pyelonephritis. Beta hemolytic streptococcus such as Streptococcus agalactiae, and S. dysgalactiae subsp. equisimilis, when grown on blood-agar, produce complete hemolysis around each colony, usually cause severe infections including many cases of cellulitis, pneumonia, meningitis and sepsis. However the analysis of pyelonephritis causing beta hemolytic streptococcus has seldom been investigated. In this study, we tried to clarify the relationship between beta hemolytic streptococcus and pyelonephritis patient in Japan. We investigated beta hemolytic streptococcus isolated from pyelonephritis patient at two tertiary care hospitals in the central region of Japan by analyzing the background and antimicrobial susceptibility of beta hemolytic streptococcus. We recovered total sixteen beta hemolytic streptococcus in nine years. In pyelonephritis patients, beta hemolytic streptococcus was isolated from 9 S. agalactiae and 7 S. dysgalactiae subsp. equisimilis, respectively. The numbers of streptococci from female patients were more than those from male patients. Bacteria were not isolated from under 30 year patients. All S. dysgalactiae subsp. equisimilis were isolated from 60-year-age patients. About one fifth of beta hemolytic streptococcus had erythromycin and minocycline-resistant ability. Four beta hemolytic streptococci that were all S. agalactiae, had levofloxacin-resistant ability. Only one S. agalactiae had trimethoprim-sulfamethoxazole-resistant ability. Ampicillin was effective against all beta hemolytic streptococci. In our results, beta hemolytic streptococcus caused pyelonephritis in aged people and the quarter of these bacteria had some antibiotics resistance.

关键词： Beta Hemolytic Streptococcus Pyelonephritis S. agalactiae S. dysgalactiae subsp. equisimilis Antimicrobial Susceptibility