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[Evaluation of antibiotics used for enterohemorrhagic Escherichia coli O157 enteritis--effect of various antibiotics on extracellular release of verotoxin].

We tested antimicrobial activities of ten oral antibiotics; ampicillin (ABPC), cefdinir (CFDN), cefaclor (CCL), fosfomycin (FOM), norfloxacin (NFLX), nalidixic acid (NA), kanamycin (KM), minocycline (MINO), doxycycline (DOXY), and tetracycline (TC) against eleven enterohemorrhagic Esherichia coli (EHEC) O157 clinical strains. Two strains were resistant to ABPC and TC. Other strains were sensitive to all the ten antibiotics. To investigate the effect of antibiotics on extracellular release of verotoxin (VT), strain EHEC TT10 was grown in 10 ml of LB containing various concentrations of the antibiotics for 2 h. Number of viable cells were counted and the amounts of VT1 and VT2 released in the supernatants were measured with reverse passive latex agglutination (RPLA) using serially diluted sterilized culture supernatants. The amount of VT1 and VT2 was evidently increased with ABPC, CFDN, CCL, and FOM, the inhibitors of cell wall biosynthesis. In the case of quinolons, VT2 was markedly increased, but VT1 was not released to the supernatant. KM killed the bacteria efficiently, but no release of VT1 or VT2 was observed in the supernatant. Tetracyclines (MINO, DOXY, and TC) did not make the bacteria release either VT1 or VT2, but could not kill the bacteria appreciably. These results indicated that the inhibitors of protein synthesis (KM, MINO, DOXY, TC) are the safe antibiotics not causing the release of verotoxin from the cells and thus preventing development of hemolytic uremic syndrome (HUS) or thrombotic thrombocytopenic purpra (TTP), the important sequelae of the enteritis.

Ampicillin Resistance↗

[In vitro specific binding of Shiga toxin 1 and 2 by TAK-751S (Gb3 analog)].

TAK-751S is a synthetic trisaccharide coupled to Chromosorb P using a spacer sequence of 8-methoxycarboyloctyl (MCO). Its chemical structure is similar to a human receptor (Gb3) of Stx produced by enterohemorrhagic Escherichia coli (EHEC). In vitro efficacy of TAK-715S was studied by using ACHN cultured cell assay, which is sensitive and specific for measuring low level of Stx. Under various conditions, TAK-715S was mixed with purified Stx1 and Stx2, and residual free toxins in the solution were measured by using ACHN cells. TAK-715S was demonstrated to bind specifically to Stx1 and Stx2 under the condition similar to a human intestine while Chromsorb P did not bind to any Stx. The binding activity was stable in the presence of various processed foods, fresh vegetables and fruits. Antibiotics such as fosfomycin, kanamycin and norfloxacin did not disturb its binding capability. Minimum inhibitory concentrations of these antibiotics against Staphylococcus aureus FDA209P or E. coli NIHJ JC-2 neither changed after incubating with TAK-751S for 60 min at 37 degrees C. These results suggest that TAK-751S can be given orally with various foods and antibiotics for the elimination of Stx1 and Stx2 in the gut of patients with EHEC infections.

Adsorption↗

[A Shigella sonnei outbreak in Nagasaki].

We have experienced an outbreak of dysentery in Nagasaki. Shigella sonnei were positively cultured from 467 patients out of suspected 821 cases, and 346 patients were admitted. 121 patients were treated with oral antimicrobials in the outpatient clinic. Five patients were diagnosed as secondary infection. We treated a total of 96 patients in Nagasaki Municipal Medical Center, and studied the clinical and bacterial features in these 96 patients. Chief complaints included fever, abdominal pain and diarrhea. Most diarrheal patients showed waterly diarrhea and only a few were bloody (3 of 47). Treatment of levofloxacine 300 mg a day for 5 days successfully eliminated S. sonnei from all culture positive patients. An environmental surveillance revealed that water in a well at the university to which many patients were using was the origin of the infection with positive cultures of S. sonnei. No difference between the clinical and environmental isolates was observed in results on biochemical, serological and enzymatic tests. All isolates were susceptible to levofloxacin and to ofloxacin, but three isolates showed resistance of fosfomycin with MIC above 64 micrograms/ml. In analysis of pulsed-field gel electrophoresis, both clinical and environmental isolates were considered to be closely related.

Anti-Infective Agents↗

[Multidrug-resistant and fluoroquinolone-resistant Salmonella enterica serotype Typhimurium definitive phage type 12 isolated from infantile diarrhea].

A 35-day-old male infant was admitted to our hospital, presenting a high fever, vomiting, and diarrhea. Multidrug-resistant and fluoroquinolon-resistant Salmonella serotype Typhimurium was isolated from his stool. The phage type of the strain was DT12. The strain was resistant to ampicillin, streptomycin, gentamicin, tetracycline, chloramphenicol, sulfamethoxazole-trimethoprim, and fluoroquinolones (levofloxacin; MIC 8 micrograms/ml, norfloxacin; 24 micrograms/ml, ciprofloxacin; 16 micrograms/ml, sparfloxacin; 32 micrograms/ml). He was cured by antibiotic therapy using fosfomycin (< or = 1 microgram/ml). We could not determine a route of infection. Domestic surveillance for fluoroquinolon-resistant Sallmonella is necessary.

Anti-Infective Agents↗

[Comparison of multidrug-resistant Salmonella enterica serovar Newport isolates from a patient and sewages in Fukui Prefecture].

From May 2003 to March 2004, nine strains of multidrug-resistant (MDR) Salmonella enterica serovar Newport were isolated from 5 sewages of 3 sewage treatment plants located in 2 different regions of Fukui Prefecture. Antimicrobial susceptibility testing identified 2 different MDR patterns (type I and II). Type I strains were resistant to ampicillin, tetracycline, streptomycin, chloramphenicol, amoxicillin/clavulanic acid, sulfisoxazole, cefotaxime, cephalothin, ceftriaxone, and ceftrazidime and type II strains were additionally resistant to sulfamethoxazole-trimethoprim and spectionomycin. Neither of kanamycin, fosfomycin, nalidixic acid, gentamycin, or ciprofloxacin resistance was observed. Type I and II strains were different in pulsed-field gel electrophoresis patterns and plasmid profiles, while they showed the same profiles with in each type. All the MDR strains in this study were proved to carry the blaCMY-2 gene by PCR and sequence analyses. Numbers of reports on MDR S. Newport with blaCMY-2 have been increasing in foreign countries such as the United States. The MDR patterns of the isolates in this study were similar to those of the United States. These properties of the type I strains were identical with that a strain isolated from a 7-year-old patient in Fukui Prefecture, in September 2003 which we previously reported. However, the patient lived in a region where no epidemiological associations with the sewage treatment plants were found. This suggests that the MDR strains might prevail in Fukui Prefecture. Domestic surveillance for MDR S. Newport would be need.

Anti-Bacterial Agents↗

Tolerant response of Streptococcus sanguis to beta-lactams and other cell wall inhibitors.

In contrast to group A streptococci or Streptococcus pneumoniae, cells of Streptococcus sanguis (group H) do not exhibit the irreversible effects of penicillin treatment, such as loss of viability or lysis. On the other hand, the same bacteria show typical effects of penicillin, such as morphological alterations, reduction in the rate of cell wall synthesis, and secretion of murein and lipoteichoic acid polymers into the medium. A novel effect of cell wall inhibitors was also noted: treatment with beta-lactams or with fosfomycin, d-cycloserine, or beta-halogeno-d-alanine caused the release of substantial amounts of glycerol lipids into the growth medium. The antibiotic "tolerance" of S. sanguis is interpreted in terms of the hypothesis that the activity of bacterial murein hydrolases is essential for the irreversible effects of cell wall inhibitors.

Alanine↗

Effects of cerulenin on antibiotic-induced lysis of streptococcus faecalis (S. faecium).

Addition of the antibiotic cerulenin to cultures lowered the minimal effective concentration of penicillin G or methicillin required to produce bacterial lysis and killing. This effect was most pronounced at subinhibitory antibiotic concentrations. Cerulenin had no significant effects on lysis or killing induced in the presence of D-cycloserine, fosfomycin, bacitracin, or vancomycin.

Anti-Bacterial Agents↗

In vitro activity of LY146032 alone and in combination with other antibiotics against gram-positive bacteria.

The antibacterial activity of LY146032 alone and in combination with other drugs was assayed against gram-positive isolates. Synergism was found when LY146032 was combined with netilmicin, amikacin, imipenem, and fosfomycin by both checkerboard and time-kill tests. Indifference predominated when LY146032 was combined with teicoplanin, vancomycin, and rifampin. Under no circumstances and with no combinations was an antagonistic effect detected.

Anti-Bacterial Agents↗

In vitro susceptibilities of Pseudomonas pseudomallei to 27 antimicrobial agents.

Clinical isolates of Pseudomonas pseudomallei isolated in Thailand from 1981 to 1989 were tested for their in vitro susceptibilities to 27 antimicrobial agents, including older and newer quinolones, broad-spectrum cephems, carbapenems, monobactams, penicillins, tetracyclines, chloramphenicol, rifamycin, sulfamethoxazole, trimethoprim, and fosfomycin. Tosufloxacin, meropenem, CS-533, and minocycline were active against P. pseudomallei at levels comparable to or even greater than those of antimicrobial agents tested in previous studies, such as ciprofloxacin, ceftazidime, imipenem, carumonam, and piperacillin. Drug-resistant P. pseudomallei was found in only 1% of the isolates. The drug-resistant P. pseudomallei isolates displayed a unique pattern of susceptibility to the above-listed drugs.

Anti-Bacterial Agents↗

In vitro activities of new antimicrobial agents against multiresistant Staphylococcus aureus isolated from septicemic patients during a Belgian national survey from 1983 to 1985.

The antimicrobial agents most active against bacteremic isolates of oxacillin-resistant Staphylococcus aureus isolated from 1983 to 1985 were new fluoroquinolones, including PD 117,596 and PD 127,391 (MIC for 90% of isolates [MIC90] in agar, in micrograms per milliliter, 0.1) and temafloxacin, pefloxacin, and ofloxacin (MIC90, 0.4). Other active antimicrobial agents included fusidic acid (MIC90, 0.2) and fosfomycin (MIC90, 12.5). Vancomycin was active against all isolates. Mupirocin was very active (MIC90, 0.4).

Anti-Bacterial Agents↗

In vitro susceptibility of Campylobacter jejuni to rokitamycin.

The susceptibilities of 100 clinical isolates of Campylobacter jejuni to rokitamycin, a new macrolide, and eight other oral antibiotics were tested by using a broth microdilution method. Rokitamycin demonstrated antibacterial activity that was higher than those of fosfomycin and the tetracyclines and comparable to those of erythromycin and clindamycin, whereas it was slightly less active than the quinolones.

Anti-Bacterial Agents↗

Susceptibilities of anaerobic bacteria isolated from animals with ovine foot rot to 28 antimicrobial agents.

The agar dilution method was used to determine the inhibitory activities of 28 antimicrobial agents against 35 strains of the genus Peptostreptococcus, 4 strains of the species Peptococcus niger, 20 strains of the species Megasphaera elsdenii, 7 strains from the species Acidaminococcus fermentans, 8 strains of the genus Clostridium, 11 strains of the genus Eubacterium, and 1 strain of the species Propionibacterium acidipropionici, all of which were isolated from 125 clinical cases of ovine foot rot between January 1987 and December 1988. The three unreidopenicillins studied proved to be the most active antimicrobial agents, with a high percentage of strains being susceptible at a concentration of 64 micrograms/ml. Penicillin G, ampicillin, and the three cephalosporins studied also had good activity. Fosfomycin showed a high degree of activity among the 116 anaerobic bacteria tested.

Animals↗

Antimicrobial susceptibilities of Corynebacterium species and other non-spore-forming gram-positive bacilli to 18 antimicrobial agents.

The susceptibilities of 265 strains of Corynebacterium species and other non-spore-forming gram-positive bacilli to 18 antimicrobial agents were tested. Most strains were susceptible to vancomycin, doxycycline, and fusidic acid. Corynebacterium jeikeium and Corynebacterium urealyticum were the most resistant organisms tested. Resistance to beta-lactams, clindamycin, erythromycin, azythromycin, ciprofloxacin and gentamicin was common among strains of Corynebacterium xerosis and Corynebacterium minutissimum. Ampicillin resistance among Listeria monocytogenes was more prevalent than previously reported. Optochin, fosfomycin, and nitrofurantoin showed very little activity against most organisms tested, but the use of nitrofurantoin as a selective agent in culture medium may prevent the recovery of some isolates. Except for the unvarying activity of vancomycin against Corynebacterium species, the antimicrobial susceptibilities of the latter to other antibiotics are usually unpredictable, such that susceptibility tests are necessary for selecting the best antimicrobial treatment.

Anti-Bacterial Agents↗

Antimicrobial susceptibilities of Stomatococcus mucilaginosus and of Micrococcus spp.

The in vitro susceptibilities of 63 isolates of Stomatococcus mucilaginosus and of 188 isolates of Micrococcus spp. to 18 antimicrobial agents were determined by the agar dilution method. Many beta-lactams, imipenem, rifampin, and the glycopeptides were shown to be active in vitro against Stomatococcus and Micrococcus isolates, whereas the activities of antibiotics such as some aminoglycosides, erythromycin, and fosfomycin against an important number of these microorganisms are limited.

Anti-Bacterial Agents↗

MICs of 28 antibiotic compounds for 14 Bartonella (formerly Rochalimaea) isolates.

We assessed in vitro the antibiotic susceptibilities of 14 Bartonella isolates of the species B. quintana, B. vinsonii, B. henselae, and B. elizabethae. Columbia agar base supplemented with 5% horse blood was used as the antibiotic assay medium. Bacterial growth could be evaluated within 5 days after incubation of the plates at 37 degrees C in a 5% carbon dioxide atmosphere. The MICs at which 90% of isolates are inhibited (MIC90s) were 0.06 microgram/ml for penicillin G and amoxicillin and 0.25 microgram/ml for ticarcillin and cefotaxime. The MIC90s of oxacillin and cephalothin were 4 and 16 micrograms/ml, respectively. The MIC90s ranged from 1 to 4 micrograms/ml for aminoglycosides. Erythromycin, doxycycline, and rifampin displayed MIC90s of 0.12, 0.12, and 0.25 microgram/ml, respectively. MIC90s were 1 and 5 micrograms/ml for trimethoprim-and sulfamethoxazole, respectively, 64 micrograms/ml for fosfomycin, and 16 micrograms/ml for colistin and vancomycin. The study confirms the high levels of in vitro susceptibility of Bartonella agents to antibiotics.

Anti-Bacterial Agents↗

Susceptibilities of fluoroquinolone-resistant strains of Campylobacter jejuni to 11 oral antimicrobial agents.

The resistance of Campylobacter jejuni strains to the fluoroquinolones is increasingly frequent, and in our area it reaches nearly 50%. We studied the susceptibilities of 60 of these strains to 11 oral antibiotics. All strains except one were susceptible to the macrolides tested, with azithromycin being the most active agent tested. Of the rest of the antibiotics studied, amoxicillin-clavulanic acid, clindamycin, and fosfomycin displayed good in vitro activities. Knowledge of the susceptibilities of these microorganisms to a varied group of oral agents is necessary in view of the appearance of multiresistant strains, such as those included in our series.

Administration, Oral↗

Novel antibiotic susceptibility tests by the ATP-bioluminescence method using filamentous cell treatment.

Antimicrobial susceptibility testing by the ATP-bioluminescence method has been noted for its speed; it provides susceptibility results within 2 to 5 h. However, several disagreements between the ATP method and standard methodology have been reported. The present paper describes a novel ATP method in a 3.5-h test which overcomes these deficiencies through the elimination of false-resistance discrepancies in tests on gram-negative bacteria with beta-lactam agents. In our test model using Pseudomonas aeruginosa and piperacillin, it was shown that ATP in filamentous cells accounted for the false resistance. We found that 0.5% 2-amino-2-methyl-1,3-propanediol (AMPD) extracted ATP from the filamentous cells without affecting normal cells and that 0.3 U of adenosine phosphate deaminase (APDase)/ml simultaneously digested the extracted ATP. We used the mixture of these reagents for the pretreatment of cells in a procedure we named filamentous cell treatment, prior to ATP measurements. This novel ATP method with the filamentous cell treatment eliminated false-resistance discrepancies in tests on P. aeruginosa with beta-lactam agents, including piperacillin, cefoperazone, aztreonam, imipenem-cilastatin, ceftazidime, and cefsulodin. Furthermore, this novel methodology produced results which agreed with those of the standard microdilution method in other tests on gram-negative and gram-positive bacteria, including P. aeruginosa, Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis, for non-beta-lactam agents, such as fosfomycin, ofloxacin, minocycline, and aminoglycosides. MICs obtained by the novel ATP method were also in agreement with those obtained by the agar dilution method of susceptibility testing. From these results, it was shown that the novel ATP method could be used successfully to test the activities of antimicrobial agents with the elimination of the previously reported discrepancies.

Adenosine Triphosphate↗