[Nationwide surveillance of parenteral antibiotics containing meropenem activities against clinically isolated strains in 2012].

The nationwide surveillance of antibacterial susceptibility to meropenem (MEPM) and other parenteral antibiotics against clinical isolates during 2012 in Japan was conducted. A total of 2985 strains including 955 strains of Gram-positive bacteria, 1782 strains of Gram-negative bacteria, and 248 strains of anaerobic bacteria obtained from 31 medical institutions were examined. The results were as follows; 1. MEPM was more active than the other carbapenem antibiotics tested against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae. MEPM was also active against most of the species tested in Gram-positive and anaerobic bacteria, except for multi-drug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA). 2. Of all species tested, there were no species, which MIC90 of MEPM was more than 4-fold higher than those in our previous studies in 2009 or 2006. Therefore, the tendency to increase in antimicrobial resistance rates was not observed. 3. MEPM resistance against Pseudomonas aeruginosa was 17.8% (56/315 strains). Compared to our previous results, it was the lowest than that in 2006 and 2009. 4. Carbapenem-resistant Klebsiella pneumoniae, and multi-drug-resistant Acinetobacter species, which emerged in worldwide, were not observed. 5. The proportion of extended-spectrum beta-lactamase (ESBL) strains was 6.2% (59/951 strains) in enterobacteriaceae, which increased compared with that of our previous studies in 2009 or before. Whereas, the proportion of metallo-beta-lactamase strains was 1.6% (5/315 strains) in P. aeruginosa, which was stable. In conclusion, the results from this surveillance suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem for serious infections treatment at present, 17 years passed after available for commercial use in Japan.

An effective active surveillance method for controlling nosocomial MRSA transmission in a Japanese hospital.

Hospital-wide active surveillance for methicillin-resistant Staphylococcus aureus (MRSA) targeted to adult patients with a history of MRSA carriage within the past 5 years was performed in Juntendo University Hospital (JUH) over a 2-year period. In the first year, MRSA screening culture was ordered by physicians in charge. In the second year, infection-control practitioners (ICPs) took samples for active surveillance culture. The average monthly transmission rate of MRSA in JUH was 0.35 per 1,000 bed-days in the first year and decreased significantly to 0.26 per 1,000 bed-days in the second year (P < 0.05). In the second year, more active commitment of ICPs to MRSA screening was effective in improving the performance rate of screening, shortening turn-around time of screening results, and decreasing transmission rate. Increasing compliance with active MRSA surveillance by involvement of ICPs, targeting patients with a previous history of MRSA carriage in the previous 5 years, was effective to control nosocomial MRSA transmission.

Analysis of Staphylococcal cassette chromosome mec in BD GeneOhm MRSA assay-negative strains.

The BD GeneOhm MRSA assay could identify methicillin-resistant Staphylococcus aureus (MRSA) strains at a high ratio (97.8%). Analysis of 11 assay-negative MRSA strains suggested that insertion of non-mec staphylococcal cassette chromosome elements (SCCs) downstream of orfX, and carriage of SCCmecs with a left extremity that cannot be detected by the kit, might lead to their being given an incorrect negative status.

[Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (2006)].

From October 2006 to September 2007, we collected the specimen from 356 patients with lower respiratory tract infections in 14 institutions in Japan, and investigated the susceptibilities of isolated bacteria to various antibacterial agents and patients' characteristics. Of 414 strains that were isolated from specimen (mainly from sputum) and assumed to be bacteria causing in infection, 407 strains were examined. The isolated bacteria were: Staphylococcus aureus 64, Streptococcus pneumoniae 96, Haemophilus influenzae 87, Pseudomonas aeruginosa (non-mucoid) 52, P. aeruginosa (mucoid) 11, Klebsiella pneumoniae 20, and Moraxella catarrhalis 44. Of 64 S. aureus strains, those with 2 microg/ml or less of MIC of oxacillin (methicillin-susceptible S. aureus: MSSA) and those with 4 microg/ml or more of MIC of oxacillin (methicillin-resistant S. aureus: MRSA) were 27 (42.2%) and 37 (57.8%) strains, respectively. Against MSSA, imipenem had the most potent antibacterial activity and inhibited the growth of all strains at 0.063 microg/ml or less. Against MRSA, vancomycin and linezolid showed the most potent activity and inhibited the growth of all the strains at 1 microg/ml. Carbapenems showed the most potent activities against S. pneumoniae and in particular, panipenem inhibited the growth of all the strains at 0.063 microg/ml or less. Imipenem and faropenem also had a preferable activity and inhibited the growth of all the strains at 0.125 and 0.5 microg/ml, respectively. In contrast, there were high-resistant strains (MIC: over 128 microg/ml) for erythromycin (45.8%) and clindamycin (20.8%). Against H. influenzae, levofloxacin showed the most potent activity and its MIC90 was 0.063 microg/ml or less. Meropenem showed the most potent activity against P. aeruginosa (mucoid) and its MIC90 was 0.5 microg/ml. Against P. aeruginosa (non-mucoid), tobramycin had the most potent activity and its MIC90 was 2 microg/ml. Against K. pneumoniae, cefozopran was the most potent activity and inhibited the growth of all the strains at 0.063 microg/ml or less. Also, all the antibacterial agents except ampicillin generally showed a potent activity against M. catarrhalis and the MIC90 of them were 2 microg/ml or less. The approximately half the number (50.6%) of the patients with respiratory infection were aged 70 years or older. Bacterial pneumonia and chronic bronchitis accounted for 49.2% and 28.1% of all the respiratory infections, respectively. The bacteria frequently isolated from the patients with bacterial pneumonia were S. pneumoniae (29.2%), S. aureus (20.8%), and H. influenzae (12.9%). H. influenzae (25.0%) and P. aeruginosa (21.7%) also were frequently isolated from the patients with chronic bronchitis. Before the antibacterial agent administration, the bacteria frequently isolated from the patients were S. pneumoniae (27.5%) and H. influenzae (22.5%). The bacteria frequently isolated from the patients treated with macrolides was P. aeruginosa, and its isolation frequently was 39.4%.

[Importance of information from the Department of Clinical Laboratory in the treatment of infectious diseases--from the viewpoint of medical safety].

For the report of microbiological laboratory staff to be made more effective, developments of the entire support systems regarding hospital infection control are essential. Therefore, it should be carried out side by side with the development of a guideline support system, proper antibiotic use, a consultation system, as well as education and training of medical staff. As measures of the Department of Clinical Laboratory, antibiograms are conducted periodically and a blood culture report is taken as a 24-hour system. In addition, the blood culture report is transmitted to ensure that the attending physician performs activities according to the electronic medical records as well as through contact by telephone. In addition, the ICD reported during the day on behalf of the laboratory technician, at the stage of the first report, and the estimation of bacterial species and suggestions for additional testing were performed. For the measures described above, the current rate of two sets of blood cultures taken comprises over 90%. In addition, the use of carbapenems was reduced by half. As the result, the rate of imipenem-resistant Pseudomonas aeruginosa was reduced by about 40% in 2006 to 20% in 2010, and the development of multi-drug resistant bacteria was markedly reduced.

Comparison of antifungal activities of gentian violet and povidone-iodine against clinical isolates of Candida species and other yeasts: a framework to establish topical disinfectant activities.

We evaluated antifungal activity as assessed by the contact time in topical use of gentian violet (GV) and povidone-iodine (PI) against Candida strains. A total of 102 yeast isolates were used in this study. A markedly lower minimal inhibitory concentration (MIC)(90) of GV than of PI was detected for all yeast isolates. No remarkable difference in the MICs was observed among the identical strains isolated from different clinical sites for both GV and PI. Although the minimal fungicidal activities (MFCs) of PI were identical for all tested time points, the fungicidal activity of GV decreased during the time course of incubation. These results indicate that, whereas GV is more effective than PI, the topical disinfectant efficacy of GV should be estimated using the MFC(5 min) and not the MIC or the MFC(24 h) for overall prevention of catheter-related bloodstream infections and oral infections.

Association between antimicrobial consumption and clinical isolates of methicillin-resistant Staphylococcus aureus: a 14-year study.

The objective of this study was to determine the relationship between clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and antimicrobial consumption in hospitalized patients over a 14-year period. The study was retrospectively conducted between January 1995 and December 2008 at Juntendo University Hospital, Tokyo, Japan, a 1,020-bed tertiary-care teaching hospital. The incidence of MRSA isolates was examined using clinical specimens presented to the microbiology laboratory in the hospital. Antimicrobial consumption through intravenous injection was calculated in terms of the number of defined daily doses per 100 bed-days. The correlation between the incidence of MRSA isolates and antimicrobial consumption was determined employing a multiple stepwise regression analysis. A total of 109,946 bacterial isolates were consecutively collected over the 14-year period, and, of these, 13,872 (64% of S. aureus strains excluding coagulase-negative staphylococci) were MRSA strains. The longitudinal observation showed that the number and rate of MRSA isolates marginally decreased. The rate of MRSA isolates among S. aureus strains in 1995 was 68.5%, whereas that in 2008 was 53.8%. Consumption of cephalosporins decreased. Among carbapenems, the rate of imipenem (IPM) consumption decreased, whereas that of meropenem increased. A multiple stepwise regression analysis revealed that the antimicrobial consumption of cefmetazole, cefotiam, and IPM was positively correlated with the incidence of MRSA isolates. The use of ß-lactam antimicrobials may contribute to the development of MRSA strains.

Heterogeneously vancomycin-intermediate Staphylococcus aureus (hVISA) emerged before the clinical introduction of vancomycin in Japan: a retrospective study.

Vancomycin-intermediate Staphylococcus aureus (VISA) and its precursor, heterogeneous VISA (hVISA), are increasingly the cause of vancomycin treatment failure. Prolonged glycopeptide treatment causes the emergence of these pathogens. However, we recently reported that hVISA can be generated by methicillin-resistant S. aureus (MRSA) exposure to imipenem (Katayama et al., Antimicrob Agents Chemother. 53:3190-6). We report here a retrospective prevalence study of VISA and hVISA on 750 MRSA clinical strains isolated from 31 Japanese national university hospitals in 1990, the year before the introduction of injectable vancomycin into clinical use in Japan in 1991. No VISA strain was identified, but population analysis identified 38 hVISA strains (5.1%) from 19 hospitals. We also determined the nucleotide sequences of vraSR, walRK, clpP, and rpoB genes whose mutations are known to be associated with vancomycin resistance. When compared with vancomycin-susceptible MRSA strain N315, six of the 38 hVISA strains possessed nonsynonymous mutations in vraSR, seven in walRK, and two in rpoB genes, Thirteen of 38 (34.2%) hVISA strains possessed at least one of these mutations. Results were consistent with our hypothesis that hVISA was present in Japanese hospitals before the clinical introduction of vancomycin.

[Nationwide surveillance of parenteral antibiotics containing meropenem activities against clinically isolated strains in 2009].

The antibacterial activity of meropenem (MEPM) and other parenteral antibiotics against clinical isolates of 2655 strains including 810 strains of Gram-positive bacteria, 1635 strains of Gram-negative bacteria, and 210 strains of anaerobic bacteria obtained from 30 medical institutions during 2009 was examined. The results were as follows; (1) MEPM was more active than the other carbapenem antibiotics tested against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae. MEPM was also active against most of the species tested in Gram-positive and anaerobic bacteria, except for multidrug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA). (2) MEPM maintained potent and stable antibacterial activity against Pseudomonas aeruginosa. The proportion of MEPM-resistant strains to ciprofloxacin-resistant strains or imipenem-resistant strains were 53.1% and 58.0% respectively. (3) The proportion of extended-spectrum beta-lactamase (ESBL) strains was 3.1% (26 strains) in enterobacteriaceae. And the proportion of metallo-beta-lactamase strains was 2.0% (6 strains) in P. aeruginosa. (4) Of all species tested, there were no species except for Bacteroides fragilis group, which MIC90 of MEPM was more than 4-fold higher than those in our previous study. Therefore, there is almost no significant decrease in susceptibility of clinical isolates to meropenem. In conclusion, the results from this surveillance study suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem for serious infections treatment at present, 14 years passed after available for commercial use in Japan.

Effect of the continuous intake of probiotic-fermented milk containing Lactobacillus casei strain Shirota on fever in a mass outbreak of norovirus gastroenteritis and the faecal microflora in a health service facility for the aged.

For conducting effective risk management in long-stay elderly people at a health service facility, we performed an open case-controlled study to evaluate the effect of the intake of probiotic-fermented milk containing Lactobacillus casei strain Shirota (LcS-fermented milk) on norovirus gastroenteritis occurring in the winter season during the intake period. A total of seventy-seven elderly people (mean age 84 years) were enrolled in the study. During a 1-month period, there was no significant difference in the incidence of norovirus gastroenteritis between the LcS-fermented milk-administered (n 39) and the non-administered (n 38) groups; however, the mean duration of fever of >37°C after the onset of gastroenteritis was 1·5 (SD 1·7) d in the former and 2·9 (SD 2·3) d in the latter group, showing a significant shortening in the former group (P < 0·05). RT-quantitative PCR analysis targeting ribosomal RNA showed both Bifidobacterium and Lactobacillus to be significantly dominant, whereas Enterobacteriaceae decreased in faecal samples from the administered group (n 10, mean age 83 years), with a significant increase in faecal acetic acid concentration. Continuous intake of LcS-fermented milk could positively contribute to the alleviation of fever caused by norovirus gastroenteritis by correcting the imbalance of the intestinal microflora peculiar to the elderly, although such consumption could not protect them from the disease.

[The number of blood culture bottle sets and the clinical significance of Staphylococcus spp. isolated from the blood culture].

PURPOSE: We investigated the numbers of blood culture and percentage for 2 or more sets in last 3 years, and analyzed the positive blood culture rates based on culture set drawing in 2007. It is considered indispensable to collect two or more sets of blood culture in order to raise positive rates and discrimination ability. However, the papers which studied discrimination ability are rare; we investigated the relation of number of sets and discrimination ability in the case of Staphylococcus spp. isolated from the blood culture bottle. MATERIALS AND METHODS: In this examination, cases which performed intravascular catheter tip cultivation simultaneously were removed. In Juntendo University Hospital, 124 Staphylococcus spp. isolated from blood culture in 2007 from January to December were examined. All isolates were categorized as true-positives, contaminants, or indeterminate. The categorical decision was made by following factors taken into account: clinical course, physical findings, laboratory data and imaging results. Two or more Infection Control Doctors (ICDs) independently judged the results, and finally, in the weekly ICD round conclusions were determined. RESULTS: In 2007, 3,674 blood-culture specimens were collected. The objects of isolated bacteria were 14.8% of Staphylococcus aureus, and coagulase-negative staphylococci (CNS) 24.7%. With S. aureus, indeterminate cases decreased from 18.9% for one-set to 4.8% for two-sets (p = 0.24). In the case of CNS, indeterminate cases decreased from 34% for one-set to 7.7% for two-sets (p<0.01). CONCLUSION: By performing two-set collection, indeterminate cases showed a significant reduction. In particular, S. aureus is a bacterium which often causes blood stream infection. In one-set drawing out, since there are 18.9% of indeterminate cases, there would be a risk of the delay of clinical judgment.

Seasonal variation in liver function tests: a time-series analysis of outpatient data.

BACKGROUND: Long-term physiological variations, such as seasonal variations, affect the screening efficiency at medical checkups. This study examined the seasonal variation in liver function tests using recently described data-mining methods. METHODS: The 'latent reference values' of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyltransferase (gammaGT), cholinesterase (ChE) and total bilirubin (T-Bil) were extracted from a seven-year database of outpatients (aged 20-79 yr; comprising approximately 1,270,000 test results). After calculating the monthly means for each variable, the time-series data were separated into trend and seasonal components using a local regression model (Loess method). Then, a cosine function model (cosinor method) was applied to the seasonal component to determine the periodicity and fluctuation range. A two-year outpatient database (215,000 results) from another hospital was also analysed to confirm the reproducibility of these methods. RESULTS: The serum levels of test results tended to increase in the winter. The increase in AST and ALT was about 6% in men and women, and was greater than that in ChE, ALP (in men and women) and gammaGT (in men). In contrast, T-Bil increased by 3.6% (men) and 5.0% (women) in the summer. The total protein and albumin concentrations did not change significantly. AST and ALT showed similar seasonal variation in both institutions in the comparative analysis. CONCLUSIONS: The liver function tests were observed to show seasonal variations. These seasonal variations should therefore be taken into consideration when establishing either reference intervals or cut-off values, which are especially important regarding aminotransferases.

[Effect of continuous probiotic fermented milk intake containing Lactobacillus casei strain Shirota on fever in mass infectious gastroenteritis rest home outbreak].

As part of medical risk management in a long-term stay facility for the elderly, we introduced probiotic fermented milk containing Lactobacillus casei strain Shirota (LcS-fermented milk) in an open case-control study of its effect of (1 bottle a day) on winter-time norovirus gastroenteritis. During the 1 month from December 1 to 31, 2006, norovirus gastroenteritis occurred in 21 (55%) of 38 cases in the nonmilk group, and in 27 (64%) of 39 cases in the milk group, showing no statistically significant difference. The mean duration of >37 degrees C fever after disease onset, however, was 2.9+/-2.3 days in the non-milk group and 1.5+/-1.7 days in the milk group, showing significant shortening (p<0.05). The duration of > 38 degrees C fever after disease onset also tended to be shorter. No significant differences were seen in age, gender, days of antipyretic use, or stool-free days. These results suggested that continuous intake of LcS-fermented milk could aggravate fever duration in elderly long-term rest home residents although it probably did not prevent noroviral gastroenteritis onset.

Cyclospora infection in an immunocompetent patient in Japan.

Cyclospora cayetanensis is a coccidian protozoa that was newly recognized in 1979 in Papua New Guinea. We report the case of a 42-year-old French man who had visited Vietnam and presented with fever and watery diarrhea that had lasted for more than 2 weeks. The patient was diagnosed with C. cayetanensis infection by examination of a stool smear using UV fluorescence microscopy. Based on this rare case, we recommend that Cyclospora infection might be considered in the differential diagnosis of traveler's diarrhea in immunocompetent patients.

[Nationwide surveillance of parenteral antibiotics containing meropenem activities against clinically isolated strains in 2006].

The antibacterial activity of meropenem (MEPM) and other parenteral antibiotics against clinical isolates of 876 strains of Gram-positive bacteria, 1764 strains of Gram-negative bacteria, and 198 strains of anaerobic bacteria obtained from 30 medical institutions during 2006 was measured. The results were as follows; 1. MEPM was more active than the other carbapenem antibiotics tested against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae. MEPM was also active against most of the species tested in Gram-positive and anaerobic bacteria, except for multi-drug resistant strains including methicillin-resistant Staphylococcus aureus. 2. As for Pseudomonas aeruginosa, all of the MEPM-resistant strains were resistant to imipenem (IPM). MEPM showed low cross-resistant rate both againt IPM-resistant P. aeruginosa (41.8%) and ciprofloxacin-resistant P. aeruginosa (33.3%). 3. The proportion of extended-spectrum beta-lactamase (ESBL) strains was 4.3% (6 strains) in Escherichia coli, 1.1% (1 strain) in Citrobacter freundii, 21.7% (5 strains) in Citrobacter koseri, 3.1% (4 strains) in Klebsiella pneumoniae, 3.3% (3 strains) in Enterobacter cloacae, 0.8% (1 strain) in Serratia marcescens, and 4.9% (2 strains) in Providencia spp. The proportion of metallo-beta-lactamase strains was 3.1% (10 strains) in P. aeruginosa. 4. Of all species tested, there were no species, which MIC90 of MEPM was more than 4-fold higher than those in our previous study. Therefore, there is almost no significant decrease in susceptibility of clinical isolates to meropenem. In conclusion, the results from this surveillance study suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem at present, 11 years after available for commercial use.

[In vitro activity of methylrosaniline chloride (gentian violet) as disinfectant against Candida spp. and Trichosporon spp. isolated from blood samples].

OBJECTIVE: Methylrosaniline Chloride (MRC) is recognized as a disinfectant, but recently is rarely used in the clinic, because of its cytotoxicity when used continuously with conventional concentrations (1% MRC). We have reported the antibacterial activity of MRC with lower concentration against Methicillin-resistant Staphylococcus aureus (MRSA). In this study, we evaluated the antifungal activity of MRC with lower concentrations. MATERIAL AND METHODS: Antifungal activities of MRC against Candida spp. and Trichosporon spp. were tested. All strains tested were isolated from 106 blood or intravenous catheter samples at Juntendo University Hospital from 1995 to 2004. Minimum inhibitory concentrations against fungi were assayed by agar dilution, under both aerobic and anaerobic conditions. RESULTS: A 0.01% or less concentration of MRC solutions showed marked antifungal activity against Candida spp. and Trichosporon spp. under aerobic or anaerobic conditions. CONCLUSION: A 0.01% or less concentration of MRC should be reevaluated for the control of fungal infection and MRSA infection control.

[Isolation rate and antifungal drug susceptibility of yeast like fungi isolated from blood or vascular catheter].

Increased resistance of strains to antifungal drugs has gained increasing attention. We studied the status of fungal isolation from blood and vascular catheters at Juntendo University Hospital from 1994 to 2002. The major fungi isolated were Candida albicans, Candida parapsilosis, Candida glabrata and Candida trophicalis, or 86% yeast-like fungi. Isolation of these fungi from vascular catheters is increasing. The effectiveness of 6 anti-fungal agents against 116 yeast-like fungi was measured by microdilution. In antifungal activity of micafungin (MCFG), MIC90, was < or = 0.03 g/mL for C. albicans, C. glabrata and C. tropicalis. MCFG showed the strongest antifungal activity among the drugs tested for above Candida spp.. Five of 37 strains of C. albicans were resistant to fluconazole (FLCZ) showing MIC > or =64 g/mL. These strains were also resistant to itraconazole (ITCZ) but MICs of MGFG, flurocytosine (5-FC) and amphotericin B (AMPH-B). Two of 38 strains of C. parapsilosis are resistant to flurocytosine (5-FC) showing MIC > or =64 g/mL. There is no resistant strain of fungi (yeast-like organisms) tested against AMPH-B. Six patients from whom resistant fungi were isolated from blood and vascular catheters have severe diseases and/or are have just undergone a major surgical operation. These results indicate that it is vital for deep mycosis to start early treatment with appropriate drugs selected based on rapid detection and identification of organisms and the drug susceptibility of organisms.

[Bacterial interaction and indirect pathogenesis of Pseudomonas aeruginosa at the growth of MRSA].

Bacterial interactions such as methicillin-resistant Staphylococcus aureus (MRSA) growth inhibition or inactivation of anti-MRSA antibiotics by Pseudomonas aeruginosa as an indirect pathogen were tested by in vitro assay. Paired strains, P. aeruginosa and MRSA, used in this experiment were isolated from 63 respiratory samples at Juntendo University Hospital from 2002 to 2003. Growth inhibitory activities against MRSA by P. aeruginosa were tested with reversed agar plate method. Inactivation of anti-MRSA antibiotics by P. aeruginosa were assayed with disk diffusion method using agar over lay technique. Fifty-six (88.9%) out of 63 samples showed the significant MRSA growth inhibitory activity by co-existed P. aeruginosa. Anti-MRSA antibiotics such as trimetoprim-sulfamethoxazole combination (ST), arbekacin (ABK) and minocycline (MINO) except Vancomycin (VCM) and Teicoplanin (TEIC) were inactivated by the co-existed P. aeruginosa. Our data suggests that P. aeruginosa may play not only as a chronic respiratory pathogen but also as an indirect pathogen. Further, the most P. aeruginosa with anti-MRSA activity isolated respiratory sample may play as a modulator of MRSA infection.

[In vitro indirect pathogenesis of Pseudomonas aeruginosa against anti MRSA chemotherapy].

In the patient with a chronic respiratory disease, both Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MRSA) are frequently detected from expectoration. Vancomycin (VCM) and arbekacin (ABK) are both recommended for the chemotherapy of MRSA infection in Japan. Minocycline (MINO) is also selected for the treatment of MRSA infection. While rifampicin (RFP) and a trimetoprim-sulfamethoxazole combination (ST) are also recommended in Europe and USA but not recommended in Japan for the chemotherapy of MRSA infection. It is pointed out that coexistence bacteria affect chemotherapy as an indirect pathogen. Not only an antibacterial action but the immunological action or the metabolic effect against chronic P. aeruginosa infection such as DPB is known by the administration of 14-membered ring macrolides including erythromycin (EM). We considered the influence of P. aeruginosa isolated with MRSA on the activity against anti-MRSA agents by the disk diffusion method with bilayer flat agar in vitro. Moreover, we also examined the influence of EM against the activity of the anti-MRSA agents when P. aeruginosa was coexistence. One strain of MRSA as an indicator strain and 100 strains of P. aeruginosa as test strains, which were obtained from clinical materials, were used for the following experiment. P. aeruginosa was streaked on to the Mueller-Hinton agar culture medium (MHA), and they incubated at 35 degrees C for 24 hours. Then, the blood agar plate was piled up, MRSA was streaked on the blood agar surface, the susceptibility test disks (VCM, ABK, MINO, RFP, ST) were put on it, and incubated at 35 degrees C for a further 24 hours. The diameter of the zone of inhibition around the susceptibility disks against MRSA was measured and compared with P. aeruginosa free experiments. The anti-MRSA activity of MINO, ST and ABK was reduced by coexistence of P. aeruginosa. In RFP and VCM, the anti-MRSA activity was reinforced by coexistence of P. aeruginosa. Although the anti-MRSA activity of ST and ABK has improved by EM addition in the MHA plates, the anti-MRSA activity has not improved in MINO. These results are suggesting that in a MRSA infection, the chemotherapy by anti-MRSA agents were affected by coexistence of P. aeruginosa as an indirect pathogen. The macrolides such as EM may be useful as a modulator for chemotherapy by ST or ABK when MRSA and P. aeruginosa are isolated at the same time from the patient.