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Comparison of the exoproducts of gram-negative bacteria by SDS-Page.

The protein exoproducts released during exponential growth of Gram-negative bacteria were analysed and compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-Page). The following bacterial strains were tested: Escherichia coli, Klebsiella pneumoniae, Citrobacter freundii, Enterobacter cloacae, Serratia liquefaciens, Serratia rubidaea, Proteus mirabilis, Proteus vulgaris, Salmonella minnesota, Pseudomonas aeruginosa and Pseudomonas fluorescens. It is demonstrated by SDS-Page that members of one species show identical protein pattern, whereas different species show besides comparable protein bands a species characteristic pattern. All members of Enterobacteriaceae were shown to release proteins whose molecular weights fell into the following size regions: Each strain was shown to synthesize a polypeptide of molecular weight 34,000 and one or more polypeptides within the molecular weight range 25,000-29,000. This profile was shown to be clearly different from that of Pseudomonas strains where 20 or more distinct polypeptides ranging from 12,500 to 160,000 Mr were detectable.

Bacterial Proteins↗

Further differentiation of Enterobacteriaceae by means of siderophore-pattern analysis.

By means of a combination of 5 siderophore bioassays using several indicator strains, genera, species and subspecies of Enterobacteriaceae can be further differentiated. Enterobactin, aerobactin and other siderophores produced can be detected. Each strain shows specific pattern which we called siderophore-pattern. It is easy to separate Morganella, Proteus, Providencia, Yersinia strains from the genera Salmonella, Shigella, Escherichia coli, Enterobacter, Citrobacter, Klebsiella, Serratia. Enterobacter agglomerans strains differ from other Enterobacter species with respect to their siderophore pattern. In Salmonella strains there are differences between the subspecies I and IV. Additionally the most strains of Salmonella subspecies I from nosocomial infections produced aerobactin, in the most cases determined by plasmids. Among Shigella strains different siderophore pattern exist according to other epidemiological markers. S. flexneri strains of serovar 6 produced contrary to the strains of other serovars enterobactin. By means of the siderophore-pattern analysis E. coli strains of serovars 01; 02; 018 can be further differentiated. E. coli 01:K1 strains containing the fimbrial antigen F11 produced aerobactin whereas the F9 strains did not. All Hafnia alvei strains produced a uniform siderophore pattern, different from all other members of the enterobacteriaceae family. With this aim Hafnia alvei strains can be easily separated under practical conditions.

Enterobacteriaceae↗

Risk factors for nosocomial infections caused by gram-negative bacilli. The Hellenic Antibiotic Resistance Study Group.

Two hundred and ninety-nine Gram-negative hospital-acquired infections from 257 patients, consecutively identified during one month (November 1992) in five hospitals in the greater Athens area, were divided into four groups on the basis of the bacterium isolated: Group 1 (Escherichia coli group) included infections owing to E. coli, Group 2 (Proteus group) consisted of infections owing to Proteus spp. and Providencia spp., Group 3 (Kiebsiella/Enterobacter group) involved infections owing to Kiebsiella spp., Enterobacter spp., Citrobacter spp. and Serratia spp. Infections owing to Pseudomonas spp. and other non-fermenters were allocated into Group 4 (non-fermenters group). The four groups were studied in relation to risk factors including the duration of hospitalization, type of ward, underlying disease, history of operation, medical procedures/devices and antimicrobial therapy. A stepwise multiple logistic regression technique (SPSS Inc) was used to analyse the data, and the three groups (the Proteus group, the Klebsiella/Enterobacter group and the non-fermenters group) were analysed separately against the E. coli group. Infections with the Kiebsiella/Enterobacter group were associated with: (a) length of hospital stay before the infection, (b) treatment with newer antibiotics, and (c) hospitalization in an intensivecare unit (ICU). Infections with non-fermenters were associated with: (a) length of hospital stay before infection, (b) a urinary catheter, (c) type of disease (chronic infection being negatively associated), (d) treatment with newer antibiotics and (e) hospitalization in an ICU. Proteus group infections were associated with (a) length of hospital stay before infection, (b) treatment with newer antibiotics and (c) operation during present hospitalization (negative association). Interestingly, no specific hospitals were identified as risk factors. Identification of patients at risk for acquiring an infection owing to a nosocomial pathogen is vital in the development of a preventive strategy for hospital-acquired infections.

Catheterization↗

The prognostic factors of adult gram-negative bacillary meningitis.

Seventy-seven patients with Gram-negative bacillary meningitis (GNBM), 57 males and 20 females, aged 17-86 years, were identified at Kaohsiung Chang Gung Memorial Hospital, over an 11-year period. Fifty-four infections were community-acquired, and 23 were nosocomial; 49 were spontaneous and 28 occurred after head surgery or neurosurgery. The organisms most frequently involved were Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Acinetobacter. Rarer pathogens included Citrobacter species, Serratia marcescens, Enterobacter cloacae, and Proteus mirabilis. All patients who did not receive appropriate antibiotic therapy died. The mortality in those treated with appropriate antibiotics was 28%. Other statistically significant prognostic factors included septic shock, initial level of consciousness, hyperosmolar hyperglycemic nonketotic coma, disseminated intravascular coagulation, high cerebrospinal fluid lactate levels and leucocytosis. In the multiple logistic regression analysis, only appropriate antimicrobial therapy and septic shock were strongly associated with mortality even after adjusting for other potentially confounding factors. Despite the high mortality, management can be improved by early diagnosis, early use of appropriate antibiotics, and correction of underlying and associated medical derangement.

Adult↗

A hospital outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae investigated by RAPD typing and analysis of the genetics and mechanisms of resistance.

Between July and September 1997 a ceftazidime- and aminoglycoside-resistant strain of Klebsiella pneumoniae infected or colonized seven patients on three paediatric wards at Guy's Hospital in London. The patients were mostly neonates or infants recovering from cardiac surgery for congenital defects. The organism was probably introduced by an asymptomatic patient from Greece and the subsequent outbreak could largely be explained by person-to-person spread on individual wards and frequent transfers of patients between wards. The outbreak was controlled by patient isolation and attention to handwashing, and there were no fatalities. The organisms were non-typeable by serology but had a characteristic RAPD profile. They produced the extended spectrum beta-lactamase SHV-5 and the aminoglycoside-modifying enzymes AAC(6') + probably AAC(3)II, encoded on a conjugative plasmid of approximately 160 kb. Two other patients had multi-resistant klebsiellas, one of them an SHV-5 producer and one a TEM-5 producer, but these could be distinguished from each other and from the outbreak strain by serological and RAPD typing and by the genetics and mechanisms of their resistances. Three other multi-resistant enterobacteria were isolated during the outbreak: an Escherichia coli that had acquired the 160 kb resistance plasmid from the epidemic klebsiella, a Citrobacter isolated from one of the patients with the klebsiella but which did not produce SHV-5, and a TEM-5-producing Enterobacter. This outbreak illustrates the importance of screening patients from high-risk areas for multiply-resistant organisms on admission, and the value of bacterial typing and analysis of resistance mechanisms to define the epidemiology of hospital infection.

Adult↗

Immunochemical analyses of serum antibodies from pig herds in a Salmonella non-endemic region.

In a large comparative survey of Danish and Swedish slaughter pig herds performed prior to this work, it was unexpectedly found that some Swedish herds harbored seropositive pigs. Serum samples from the Swedish herds had moderate responses in the Salmonella mix-ELISA (detecting serogroup B and C1 infections) compared to the Danish herds classifying some of them as seropositive using a cut-off value at 40 OD%. In Sweden, extensive Salmonella control is carried out by bacteriological screening of feces and lymph nodes, and the overall prevalence has been proven to be below 0.1%. The serological positive results were therefore unexpected; hence the reactivities of the Swedish sera were studied by a number of immunochemical analyses (Western blot, indirect ELISA, inhibition ELISA, avidity ELISA) and compared to sera from Danish pig herds with verified Salmonella infections ("the reference sera"). In Western blot, the Swedish sera had high binding reactivities against Salmonella Typhimurium LPS of different molecular weights, and gave binding patterns similar to that of the reference sera. Pre-incubation with free S. Typhimurium LPS or PS (the polysaccharide part of LPS) was able to inhibit the reactivity of the Swedish sera in the mix-ELISA. Reactivities against other related bacterial LPS such as Citrobacter freundii LPS and Yersinia enterocolitica O:3 LPS were observed in the Swedish sera, but these LPS antigens were unable to inhibit the reactivities in the mix-ELISA as efficiently as S. Typhimurium LPS. Furthermore, the Swedish sera did not bind Salmonella LPS of another serogroup (S. Meleagridis LPS, serogroup E1) or rough Salmonella LPS, both lacking the specific O-antigenic parts of S. Typhimurium LPS. The avidity of the Swedish sera was much lower than the avidity of the reference sera, which could indicate the presence of transient low-dose infections or stimulation by inactivated bacteria in feed. The results obtained in this investigation strongly indicate that the Swedish sera contain antibodies directed against the O-antigenic part of LPS from S. Typhimurium or possibly on as yet unknown bacterium.

Animals↗

Development of an ELISA using a recombinant 41 kDa partial protein (P45N') for the detection of Riemerella anatipestifer infections in ducks.

Riemerella anatipestifer, a gram-negative bacillus, is the causative agent of duck septicemia, a disease which could incur much economic loss in the duck industry. An indirect enzyme-linked immunosorbent assay (ELISA) has been developed to facilitate early detection of R. anatipestifer infection in ducks. The antigen used was a recombinant 41 kDa N-terminal fragment (rP45N') of a newly characterized R. anatipestifer potential surface protein, P45, which was expressed in Escherichia coli as an N-terminal GST fusion protein. The rP45N'-based ELISA successfully detected P45 antibodies in the sera of 20 ducks immunized with bacterin preparations of R. anatipestifer serotypes 1, 10 15, 19 and the ATCC11845 strain. Antibodies to P45 were also detected in the sera of 25% (75/296) of White Pekin ducks which were imported into Singapore from three different farms. Successful discrimination was obtained between sera from infected ducks and that of specific-pathogen free ducks (p<0.01). The rP45N'-GST antigen did not cross-react with antibodies in sera from guinea pigs which were infected with other gram-negative and gram-positive bacterial pathogens, including Aeromonas hydrophila, Citrobacter freundii, E. coli, Klebsiella pneumoniae, Pastuerella multocida, Proteus mirabilis, Salmonella spp., Serratia maccescens, Shigella sonnei and Yersinia enterocolitica. In addition, the DNA sequence encoding P45 was detected in R. anatipestifer serotypes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 14, 15, 16, 17, 18, 19 and the ATCC11845 strain, suggesting that P45 is probably also universally expressed in these R. anatipestifer serotypes. Thus, the ELISA described is applicable to the detection of R. anatipestifer infection in ducks.

Animals↗

Head-space gas-liquid chromatography for the rapid laboratory diagnosis of urinary tract infections caused by enterobacteria.

Urine specimens were analysed in parallel in a hospital laboratory by routine methods which were regarded as the standard of correct diagnosis and by the rapid test already developed in a research laboratory. Technical modifications made to the rapid test ensured that its results agreed with those from routine methods and increased its rapidity so that a result was possible 4 h after receipt of the specimen. When 382 urine specimens were analysed by the modified test which is described in detail, there were neither false negatives nor false positives for infections with Escherichia, Klebsiella, Citrobacter or Proteus species.

Chromatography, Gas↗

Phylogenetic position of phytopathogens within the Enterobacteriaceae.

The almost complete 16S rDNA sequences of twenty nine plant-associated strains, representing species of the genera Erwinia, Pantoea and Enterobacter were determined and compared with those of other members of the Enterobacteriaceae. The species of the genus Erwinia may be divided into three phylogenetic groups. Cluster I represents the true erwinias and comprises E. amylovora, E. mallotivora, E. persicinus, E. psidii, E. rhapontici and E. tracheiphila. We propose to unite the species of cluster II, E. carotovora subsp. atroseptica, E. carotovora subsp. betavasculorum, E. carotovora subsp. carotovora, E. carotovora subsp. odorifera, E. carotovora subsp. wasabiae, E. cacticida, E. chrysanthemi and E. cypripedii in the genus Pectobacterium respectively as P. carotovorum subsp. atrosepticum comb. nov., P. carotovorum subsp. betavasculorum comb. nov., P. carotovorum subsp. carotovorum comb. nov., P. carotovorum subsp. odoriferum comb. nov., P. carotovorum subsp. wasabiae comb. nov., P. cacticidum comb. nov., P. chrysanthemi and P. cypripedii. The species E. alni, E. nigrifluens, E. paradisiaca, E. quercina, E. rubrifaciens and E. salicis, comprising cluster III, are being classified into a new genus Brenneria gen. nov. respectively as B. alni comb. nov., B. nigrifluens comb. nov., B. paradisiaca comb. nov., B. quercina comb. nov., B. rubrifaciens comb. nov. and B. salicis comb. nov. The species of the genus Pantoea, included in this study, form a monophyletic unit (cluster IV), closely related with Erwinia, whereas the three phytopathogenic species of the genus Enterobacter are scattered among the genera Citrobacter and Klebsiella.

Bacterial Typing Techniques↗

Cefditoren activity against nearly 1000 non-fastidious bacterial isolates and the development of in vitro susceptibility test methods.

Cefditoren (formerly ME-1206) is an investigational, orally administered cephalosporin ester with bactericidal activity against many Gram-positive and -negative organisms. Cefditoren potency against nearly 1000 non-fastidious species was determined by National Committee for Clinical Laboratory Standards (NCCLS) reference broth microdilution and standardized disk diffusion methods. Against staphylococci, usable cefditoren activity was completely correlated with oxacillin with respect to potency and susceptibility interpretation (mec A-negative strains). Cefditoren was very active against Klebsiella spp., Proteus mirabilis, Salmonella spp., and Escherichia coli (MIC(90) range, 0.12-1 microg/ml; median zone, 23-26 mm). Cefditoren had more limited activity against Citrobacter spp., Enterobacter spp., Serratia marcescens, and indole-positive Proteae (MIC(50) range, 0.12-1 microg/ml; MIC(90), > 16 microg/ml; median zone, 18-25 mm). Against Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Acinetobacter spp., and other non-fermentors, cefditoren was inactive (MIC(90), > 16 microg/ml; zone, 6 mm). Pharmacokinetic analysis of cefditoren showed that utilized dosages produce a plasma concentration that exceeds 0.5 microg/ml for 5 to 8 h and 1 microg/ml for 4 to 6 hours (T(1/2) ranges from 1.5-2 h). The following interpretive criteria were suggested: </= 2 microg/ml or >/= 15 mm (susceptible) and >/= 8 microg/ml or </= 11 mm (resistance) that yielded an intermethod categorical agreement of 95.8% and very major or major error rates of 0.7% and 0.3%, respectively. Alternatively, </= 1 microg/ml or >/= 18 mm (susceptible) and >/= 4 microg/ml or </= 14 mm (resistant) breakpoints resulted in 96.2% accuracy and combined serious errors of only 1.1%. Cefditoren was observed to be a very active cephalosporin ranking among the most potent available orally active beta-lactams for use against a wide variety of pathogens.

Cephalosporins↗

Comparative in vitro activity of gemifloxacin, ciprofloxacin, levofloxacin and ofloxacin in a North American surveillance study.

The in vitro activity of gemifloxacin, a new fluoroquinolone, was compared to three marketed fluoroquinolones; ciprofloxacin, levofloxacin and ofloxacin against over 4,000 recent clinical isolates covering 29 species isolated in the United States and Canada between 1997-1999. Based on MIC(90)s, gemifloxacin was the most potent fluoroquinolone tested against a majority of Gram-positive isolates: Streptococcus pneumoniae, penicillin resistant S. pneumoniae, macrolide resistant S. pneumoniae, ciprofloxacin non-susceptible (MIC > or = 4 microg/mL) S. pneumoniae, S. pyogenes, S. agalactiae, viridans streptococci, Enterococcus faecalis, methicillin susceptible Staphylococcus aureus, methicillin resistant S. aureus, S. epidermidis, S. hemolyticus, and S. saprophyticus. Against Enterobacteriaceae and aerobic non-Enterobacteriaceae Gram-negatives, gemifloxacin was usually comparable to ciprofloxacin and levofloxacin and more potent than ofloxacin for the following species: Citrobacter freundii, Enterobacter aerogenes, E. cloacae, Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Morganella morganii, Proteus mirabilis, P. vulgaris, Providencia stuartii, Serratia marcescens, Acinetobacter lwoffii, A. baumannii, Burkholderia cepacia, Haemophilus influenzae, H. parainfluenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. Gemifloxacin was generally 16-64 fold more potent than the other fluoroquinolones tested against Gram-positive organisms and retains excellent activity comparable with ciprofloxacin and levofloxacin against a majority of Gram-negative pathogens.

Anti-Infective Agents↗

Antimicrobial resistance trends in medical centers using carbapenems: report of 1999 and 2000 results from the MYSTIC program (USA).

Trends in susceptibility to antimicrobials were assessed from United States participants using 4488 isolates in the MYSTIC Program, 1999 (10 centers) through 2000 (15 centers). Diverse types of hospitals (general service, university, cancer, federal, pediatric, cystic fibrosis) were enrolled from 13 states. In 2000, oxacillin-susceptible staphylococci were 100% susceptible to meropenem, imipenem, and cefepime; but only 88% of strains were susceptible to ceftazidime. Among Enterobacteriaceae, > or = 96% of Enterobacter spp., Citrobacter spp., and Serratia spp. were susceptible to meropenem, imipenem, and cefepime; but ceftazidime, ceftriaxone, and piperacillin/tazobactam had 5-20% resistance rates. Extended-spectrum beta-lactamase resistance rates in Klebsiella spp. and Escherichia coli (6-7% and 3-5%, respectively) were stable over 2 years. Acinetobacter spp. were 78-81% susceptible to carbapenems but only 63-72% susceptible to ciprofloxacin. Meropenem, tobramycin, and piperacillin/tazobactam were the most active against Pseudomonas aeruginosa, but ciprofloxacin inhibited only 74% in 2000. Overall, meropenem remained the most potent agent, especially against ceftazidime- or piperacillin/tazobactam-resistant strains.

Academic Medical Centers↗

Potency and antimicrobial spectrum update for piperacillin/tazobactam (2000): emphasis on its activity against resistant organism populations and generally untested species causing community-acquired respiratory tract infections.

The in vitro activity of piperacillin/tazobactam and several comparison broad-spectrum compounds was assessed against recent clinical isolates of Gram-positive and -negative bacteria from geographically diverse medical centers in Europe, North and Latin America participating in various surveillance programs in 2000. Several organisms were characterized for phenotypic expression of various resistant determinants such as extended-spectrum beta-lactamase (ESBL) or amp C cephalosporinase hyperproduction, and vancomycin resistance in enterococci (VRE). Piperacillin/tazobactam retained activity (MIC50) against oxacillin-susceptible Staphylococcus spp. (0.12-0.5 microg/ml), Bacillus spp. (0.5 microg/ml), vancomycin-susceptible enterococci (>4 microg/ml), and Corynebacterium spp. (2 microg/ml; not including C. jeikeium) with susceptibility rates of 100.0, 91.7, 85.7 and 81.8%, respectively. Piperacillin/tazobactam inhibited all Streptococcus spp. strains at < or = 16 microg/ml, including penicillin-resistant strains many of which were co-resistant to erythromycin (90%) and other beta-lactams. A specific breakpoint for these streptococci when testing piperacillin/tazobactam appears needed to prevent false-resistant reports using penicillin as a class representative. The carbapenems among beta-lactams were the most active agents against the ESBL-producing species of Escherichia coli and Klebsiella pneumoniae and those strains which hyper-express amp C enzymes including Citrobacter spp. and Enterobacter spp. Piperacillin/tazobactam only exhibited modest activity against the "amp C resistance group" strains (68.8% susceptible or intermediate, MIC < or = 64 microg/ml). Piperacillin/tazobactam (MIC50, 8 microg/ml; 79.5% susceptible) was the most active agent tested against multi-drug resistant isolates of Pseudomonas aeruginosa. Against sampled Haemophilus influenzae (39.2% ampicillin-resistant), piperacillin/tazobactam (MIC(90,) < or = 0.06 microg/ml), ceftriaxone and ceftazidime inhibited 100.0% of the isolates at < or = 0.25 microg/ml. These in vitro surveillance results from the year 2000 on three continents, demonstrated a sustained potent activity of piperacillin/tazobactam against selected problematic nosocomial and community-acquired pathogens. The potential importance of these findings is that this beta-lactamase inhibitor combination can be used an empiric treatment of serious infections in hospital environments where resistance has emerged, as well as covering nearly all isolates of fastidious respiratory tract pathogens acquired in the community setting.

Community-Acquired Infections↗

Susceptibility patterns of orally administered antimicrobials among urinary tract infection pathogens from hospitalized patients in North America: comparison report to Europe and Latin America. Results from the SENTRY Antimicrobial Surveillance Program (2000).

Urinary tract infections (UTIs) remain a worldwide nosocomial infection problem. Geographic variations in pathogen occurrence and susceptibility profiles require monitoring to provide information to guide new (garenoxacin [BMS284756]) therapeutic options. Two thousand seven hundred-eighty UTI isolates from Europe (n = 783), Latin America (531), and North America (1,466) were tested and compared against 44 agents by reference methods in the SENTRY Antimicrobial Surveillance Program. The top seven pathogens accounted for 90% of all isolates and the rank order for all regions was: Escherichia coli (1,316; 47%), Enterococcus spp. (351; 13%), Klebsiella spp. (306; 11%), Pseudomonas aeruginosa (210; 8%), Proteus mirabilis (145; 5%), Enterobacter spp. (97; 4%), and Citrobacter spp. (78; 3%). The pathogen rank order was similar among regions except for the rarer occurrence of Enterococcus spp. (Rank #6, 4%) in Latin America. E. coli ampicillin resistance was highest in Europe and Latin America (51-55%). Ampicillin (37%), ciprofloxacin or garenoxacin (4%), and trimethoprim/sulfamethoxazole (23%) resistance remained lowest in North America. Nitrofurantoin susceptibility in E. coli was still at acceptable levels and ranged from 91 to 96% across regions. The regional ciprofloxacin-resistant rank order for P. aeruginosa by region was: Latin America (55%) > Europe (41%) > North America (29%). Vancomycin-resistant enterococci (VRE) were only detected in North America (7%). Garenoxacin possessed a 34 to 44% wider spectrum compared to ciprofloxacin against enterococci UTI isolates. Extended spectrum beta-lactamase rates for E. coli and Klebsiella spp. were 4 and 19%, respectively. These results emphasized the need to assess the often striking differences in pathogen occurrence and resistance rates among the commonly encountered UTI pathogens.

Administration, Oral↗

Antibacterial activity of amifloxacin (WIN 49, 375), a new quinolone agent.

The in vitro activity of amifloxacin, a quinolone antimicrobial agent was compared with those of ciprofloxacin, enoxacin, ofloxacin and norfloxacin against gram-positive and gram-negative bacteria. Ninety percent of Escherichia coli, Klebsiella species, Aeromonas, Salmonella, Shigella, Citrobacter, Enterobacter species, Proteus mirabilis, Serratia marcescens, and Morganella morganii were inhibited by less than or equal to 0.5 microgram/ml. Amifloxacin inhibited Branhamella, Haemophilus, and Neisseria at less than or equal to 0.25 microgram/ml, and 90% of Pseudomonas aeruginosa, including gentamicin- and carbenicillin-resistant isolates, at 4 micrograms/ml. It also inhibited staphylococci, including methicillin-resistant isolates, but was less active against streptococci and Bacteroides species. Amifloxacin had in vitro activity similar to enoxacin, ofloxacin, and norfloxacin, but was less active than ciprofloxacin. Like other quinolones, it was less active at acid pH and in the presence of urine.

Anti-Bacterial Agents↗

Multicenter evaluation of antimicrobial resistance to six broad-spectrum beta-lactams in Colombia using the Etest method. The Colombian Antimicrobial Resistance Study Group.

The need for comprehensive and quantitative accurate antimicrobial resistance surveillance systems has become acute as a guide to problem recognition and to focus local interventions. A multilaboratory (10 medical centers) Colombia surveillance project was initiated in early 1997 to monitor the potency and spectrum of six (cefepime, cefotaxime, ceftazidime, cefoperazone/sulbactam, aztreonam, and imipenem) broad-spectrum antimicrobial agents tested against 100 organisms per participant center (802 strains). Ten groups of organisms were tested by a reference-quality method (Etest; AB BIODISK, Solna, Sweden) with results validated by concurrent quality control and additional challenge strain analysis. Results from nine qualifying medical centers were tabulated, and 95.7 to 96.8% of quality assurance tests were within expected ranges. Only cefepime (90.1-100.0% susceptible) and imipenem (96.3-100.0%) were active against all Enterobacteriaceae at > 90% of susceptible isolates using the breakpoint concentrations recommended by the National Committee for Clinical Laboratory Standards. Among ceftazidime- (or cefotaxime- or aztreonam-) resistant Enterobacter spp. and Citrobacter freundii, cefepime remained active, but not cefoperazone with sulbactam. Escherichia coli and Klebsiella spp. strains having resistance phenotypes consistent with extended spectrum beta-lactamase production were discovered in approximately 5 to 10% of isolates. All tested drugs except ceftazidime (31.8-57.7% susceptible) were active against > 94% of oxacillin-susceptible staphylococci. Similar rates of resistance (9.1-14.8%) were observed in Pseudomonas aeruginosa for five of six drugs (not cefotaxime; 15.9% of strains were susceptible). Acinetobacter spp. isolates were most susceptible to imipenem (95.8%), cefepime (86.1%), and cefoperazone/sulbactam (83.3%). Overall for the 1997 order of antimicrobial spectrums for these tested compounds was: imipenem (96.6%) > cefepime (93.6%) > cefoperazone/sulbactam (90.5%) > cefotaxime (74.9%) > aztreonam (74.3% for Gram-negative bacilli only) > ceftazidime (73.2%). These data should be used to guide empiric regimens in Colombia, and additionally will provide a resistance statistical baseline to which future studies in this nation can be compared.

Anti-Bacterial Agents↗

Multicenter evaluation of the in vitro activity of six broad-spectrum beta-lactam antimicrobial agents in Puerto Rico. The Puerto Rico Antimicrobial Resistance Study Group.

The minimum inhibitory concentrations of 6 broad-spectrum beta-lactam antimicrobial agents were determined by use of the Etest versus a total of 569 bacteria in 7 Puerto Rican hospital laboratories. These included 342 recent clinical isolates of Enterobacteriaceae, 63 Pseudomonas aeruginosa, 54 Acinetobacter species, and 110 oxacillin-susceptible staphylococci. Extended spectrum beta-lactamase production was noted among 11% of Klebsiella pneumoniae isolates. Hyperproduction of Amp C cephalosporinase was observed with > 20% of isolates of Enterobacter spp., Serratia spp., and Citrobacter freundii. The overall rank order of activity of the six beta-lactams examined in this study versus all clinical isolates was imipenem (95.8% susceptible) > cefepime (91.1%) > piperacillin/ tazobactam (82.3%) > cefotaxime (77.6%) > piperacillin (72.5%) > ceftazidime (67.0%).

Anti-Bacterial Agents↗

Antimicrobial activity and spectrum investigation of eight broad-spectrum beta-lactam drugs: a 1997 surveillance trial in 102 medical centers in the United States. Cefepime Study Group.

Because antimicrobial agents become less effective after the emergence of resistance mechanisms in clinically prevalent pathogens, physicians must utilize local, regional, and national antimicrobial susceptibility surveillance data to assist in choices of appropriate agents. An investigation of the spectrum and potency of eight broad-spectrum beta-lactam drugs (cefepime, cefotaxime, ceftazidime, ceftriaxone, imipenem, piperacillin with or without tazobactam, and ticarcillin/clavulanic acid) was performed using a common protocol and method (Etest; AB BIODISK, Solna, Sweden) in 102 clinical microbiology laboratories in the United States. A total of 9777 strains of Gram-negative bacilli were tested from late 1996 through April 1997. Quality assurance measures using three control strains observed quality control failures in 13 laboratories (usually ticarcillin/clavulanic acid or piperacillin), but only 2% of results required deletion. A total of 33.4% of Enterobacter spp. (1977 strains) were either resistant or intermediately susceptible to ceftazidime. Only imipenem (99.6% susceptible) and cefepime (99.1%) remained highly active against strains of Enterobacter, as well as Citrobacter freundii, indole-positive Proteae, and Serratia spp. Ceftazidime-resistant Escherichia coli and Klebsiella pneumoniae were detected at rates of 10.3% and 23.8%, respectively. Although these were participant-selected strains, only imipenem and cefepime had broad-spectrum coverage (> or = 97.1%) against these extended-spectrum beta-lactamase phenotypes. A dominant number of these extended-spectrum beta-lactamase phenotypes were reported from medical centers in the Northeast, but a nationwide distribution was observed. Among the nonenteric Gram-negative bacilli (4057 strains), the rank order of susceptibility (percent inhibited at published breakpoint concentrations) was: imipenem (86.1%) > piperacillin/tazobactam (80.1%) > cefepime (77.1%) > ceftazidime = piperacillin (74.9%) > ticarcillin/clavulanic acid (61.6%) > cefotaxime (18.2%) > ceftriaxone (12.9%). The cephalosporins, cefepime and ceftazidime, had rates of resistance for the 3005 Pseudomonas aeruginosa isolates of 10.1% and 14.4%, respectively. For all Gram-negative strains tested, only two contemporary beta-lactam antimicrobials exhibited > 90% inhibition of strains, imipenem at 93.6% and cefepime at 90.2%. These drugs were superior to the other tested compounds (48.8-84.3%). Ticarcillin/clavulanic acid had the narrowest spectrum of activity (48.8% of isolates susceptible). These results indicate that carbapenems and a new fourth-generation cephalosporin, cefepime, possess usable in vitro potencies against current clinical strains of Gram-negative bacilli, many of which harbored resistance to other antimicrobial agents.

Anti-Bacterial Agents↗