Clinical and bacteriological study of nosocomial infections due to Enterobacter aerogenes resistant to imipenem.

Enterobacter aerogenes strains resistant to imipenem were isolated in 10 patients, 7 of whom had received imipenem-cilastatin. The strains were differentiated by biotype, antibiotype, and plasmid content. All of the strains overproduced a chromosomal cephalosporinase and lost a major outer membrane protein with a size of about 40 kDa. In 5 of the 10 patients, E. aerogenes strains resistant to extended-spectrum cephalosporin were isolated during the same stay. In three patients, the similarity between the imipenem-susceptible and -resistant strains suggests the occurrence of mutation and reversion in vivo. The combination imipenem-cilastatin has been critically important for use with multiresistant strains of Enterobacter spp., but its use increases the risk of selection of imipenem-resistant strains.     

Mechanism of imipenem resistance acquired by threePseudomonas aeruginosa strains during imipenem therapy

Imipenem sensitive pretherapy isolates (MICs 1–2 mg/l) and the corresponding resistant posttherapy isolates (MICs 16 mg/l) ofPseudomonas aeruginosa from three patients undergoing imipenem treatment were analyzed to establish the resistance mechanism. The identity of pyocin types, serotypes, DNA restriction endonuclease profiles and plasmid profiles strongly suggested isogenicity of pre- and posttherapy isolates. The imipenem resistant posttherapy isolates showed cross-resistance only to another carbapenem, meropenem. There were neither qualitative nor quantitative differences between pre- and posttherapy isolates in -lactamase production. Affinity of the penicillin-binding proteins 1A, 1B, 2, 3, 4, 4 and 5 for [¹⁴C]imipenem was the same in pre- and posttherapy isolates. One-dimensional and two-dimensional gel electrophoresis of outer membrane protein preparations showed diminished expression of an outer membrane protein of about 46.5 and 47.5 kilodaltons, respectively, in the posttherapy isolates. This protein had an apparent isoelectric point of about pH 5.2 in two-dimensional gel electrophoresis. Growth in proteose peptone no. 2 broth did not reduce expression of this outer membrane protein, which spoke against its identity with the outer membrane protein D1. The permeability of the outer membrane for imipenem was reduced in the posttherapy isolates, since addition of 0.5 or 0.25 of the MIC of the permeabilizing agent ethylenediaminetetraacetate reduced the MICs of imipenem for all isolates from each patient to the same (susceptible) level. The diminished expression of one of the outer membrane proteins might be the reason for this reduced permeability.     

Resistance to methicillin and other antimicrobials among community-acquired and nosocomial Staphylococcus aureus strains in a pediatric teaching hospital in Salvador, Northeast Brazil

To report the frequency of methicillin-resistant Staphylococcus aureus (MRSA) infection and to compare the antimicrobial resistance patterns between community-acquired (CA) and nosocomial (NI) strains stratified for resistance to methicillin, this retrospective study on patients under 20 years of age was conducted from April 1995 to December 2005 in a pediatric teaching hospital in Salvador, Brazil. Of 308 S. aureus strains isolated, 185 (60.1%) were reviewed, out of which 125 (67.6%) and 55 (29.7%) had CA or NI infection, respectively, and 5 were defined as colonization. Out of the nine patients with MRSA initially diagnosed as CA, three were excluded from the analysis because of report of hospitalization during the previous year. Resistance to methicillin was more frequent among NI (30.9% vs. 4.9%, p<0.001). Resistance to other antimicrobials was more common among NI-MRSA compared with CA-MRSA. Although at a low rate, CA-MRSA has occurred among children, in this region.     

Resistance to rifampicin and isoniazid in strains of Mycobacterium tuberculosis.

Drug susceptibility studies on strains of Mycobacterium tuberculosis isolated from widely different populations of patients and tested by two different techniques indicated that all 55 strains resistant to rifampicin were also resistant to isoniazid, while many strains resistant to isoniazid were found to be susceptible to rifampicin. This observation, which has as yet unknown laboratory and clinical significance, may be particularly useful in management of patients. Further studies are called for to establish this relation.     

Beta-lactamases of type culture strains of the Bacteroides fragilis group and of strains that hydrolyse cefoxitin, latamoxef and imipenem

Susceptibilities to beta-lactam antibiotics and beta-lactamase content of two groups of Bacteroides strains were compared. Type cultures produced low levels of beta-lactamase and were susceptible to cefoxitin, latamoxef, imipenem and the combination of benzylpenicillin and clavulanic acid. Other Bacteroides strains that produced higher levels of beta-lactamase were generally less susceptible to these antibiotics; this resistance was more closely related to enzyme type than to the amount of enzyme present. The beta-lactamases produced by the test strains fell into three broad groups on the basis of antibiotic degradation and inhibitor profiles: those that inactivated benzylpenicillin, but not cefoxitin, latamoxef or imipenem, and were susceptible to inhibition by beta-lactamase inhibitors; those that hydrolysed benzylpenicillin, cefoxitin and latamoxef, but not imipenem, and which were less susceptible to inhibition by beta-lactamase inhibitors; an enzyme that inactivated all the antibiotics and was not inhibited by beta-lactamase inhibitors.     

Use of molecular methods to characterizeMoraxella catarrhalis strains in a suspected outbreak of nosocomial infection

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of whole cell protein, immunoblotting with normal human serum and restriction endonuclease analysis usingTaq I enzyme were applied to 38 clinically significant isolates ofMoraxella (Branhamella) catarrhalis obtained during a suspected outbreak of nosocomial infection. Each of 18 strains had individual profiles by at least two of the three methods (unique strains). The remaining 20 strains were assigned to five groups (A-E) on the basis of similarity by at least two of the three methods. Isolates within groups A, D and E were homologous by all three methods. Immunoblot groups B and C had two distinct whole cell protein profiles (B1 and B2) but indistinguishable restriction endonuclease profiles (group B/C). This emphasizes the need to use more than one technique in characterizing strains from suspected outbreaks of nosocomial infection. Grouped strains were more likely to originate from the same hospital ward than unique strains and were associated with a significantly longer median time from patient admission to strain isolation (14 versus 3.5 days, p<0.005). Furthermore, the -lactamase activity was homologous within the groups. The results suggest that nosocomial infection involving several distinctMoraxella catarrhalis strains persisted over a period of months, involving at least 20 patients on three different wards. Such infection is probably common in wards harbouring suitably predisposed patients. The mode of transmission remains to be elucidated, but the above three techniques possess sufficient reproducibility and discriminatory ability to constitute suitable investigative tools.     

Post-antibiotic effect of imipenem, amikacin and ciprofloxacin against various strains of Serratia marcescens

The authors compared the post-antibiotic effect (PAE) of imipenem, amikacin, ciprofloxacin, and latamoxef against Serratia marcescens ATCC 13880 (type strain) and against 12 clinical strains belonging to Grimont's most frequent biotypes: A2a, A3a, A3b, A4a, A4b, A5, A6a, A8a, A8b, A8c, TT, TCT. PAE was determined by measuring bacterial growth kinetics after one hour exposure to concentration of 2 x MIC of 10(6) CFUs in Mueller-Hinton broth. Drug removal was by 10-3 dilution of the exposed culture. A PAE was consistently present with imipenem (range 0.8-2.9 hrs), amikacin (range 1.0-4.9 hrs), ciprofloxacin (range 1.4-2.8 hrs). The duration of PAE did not correlate with MIC or Grimont's biotypes.     

Resistance to antibiotics of vibrio cholerae strains isolated in Angola]

Among 87 strains of Vibrio choleare (78 Ogawa serotype and 9 Inaba serotype strains) isolated in Angola in 1987-1990, 86% exhibited multiple resistance to antimicrobials. Eighty-four to 86% of strains were resistant to ampicillin with beta-lactamase production (MIC greater than or equal to 512 mg/l), streptomycin (MIC greater than or equal to 64 mg/l), spectinomycin (MIC greater than or equal to 1,024 mg/l), and trimethoprime-sulfisoxazole (MIC greater than 1,024 mg/l). Seventy-four per cent of strains were resistant to kanamycin (MIC = 512 mg/l), 26% to chloramphenicol (MIC = 32 mg/l), 10% to tetracycline (MIC = 16 mg/l), and 10% to gentamycin (MIC greater than or equal to 32 mg/l). Transfer to E. coli K12 was associated with a substantial increase in expression of resistance to tetracycline and chloramphenicol (CAT type I), with MICs in the 128-512 mg/l range. Transfer rates to E. coli K12 of plasmids for the various resistance phenotypes were 10(-6)/10(-8). The size of the isolated plasmids was 100 Md in diameter and belonged to the incompatibility group inc 6-C.     

Clonal diversity of nosocomial epidemic Acinetobacter baumannii strains isolated in Spain

Acinetobacter baumannii is one of the major pathogens involved in nosocomial outbreaks. The clonal diversity of 729 epidemic strains isolated from 19 Spanish hospitals (mainly from intensive care units) was analyzed over an 11-year period. Pulsed-field gel electrophoresis (PFGE) identified 58 PFGE types that were subjected to susceptibility testing, rpoB gene sequencing, and multilocus sequence typing (MLST). All PFGE types were multidrug resistant; colistin was the only agent to which all pathogens were susceptible. The 58 PFGE types were grouped into 16 clones based on their genetic similarity (cutoff of 80%). These clones were distributed into one major cluster (cluster D), three medium clusters (clusters A, B, and C), and three minor clusters (clusters E, F, and G). The rpoB gene sequencing and MLST results reflected a clonal distribution, in agreement with the PFGE results. The MLST sequence types (STs) (and their percent distributions) were as follows: ST-2 (47.5%), ST-3 (5.1%), ST-15 (1.7%), ST-32 (1.7%), ST-79 (13.6%), ST-80 (20.3%), and ST-81 (10.2%). ST-79, ST-80, and ST-81 and the alleles cpn60-26 and recA29 are described for the first time. International clones I, II, and III were represented by ST-81, ST-2, and ST-3, respectively. ST-79 and ST-80 could be novel emerging clones. This work confirms PFGE and MLST to be complementary tools in clonality studies. Here PFGE was able to demonstrate the monoclonal pattern of most outbreaks, the inter- and intrahospital transmission of bacteria, and their endemic persistence in some wards. MLST allowed the temporal evolution and spatial distribution of Spanish clones to be monitored and permitted international comparisons to be made.     

Resistance of intestinal strains of Bacteroides to antibiotics and chemotherapeutic agents]

Antibacterial therapy of anaerobic infections usually involves chemotherapy. The basis of therapy is assessment of the minimal inhibitory concentration (MIC) of antibiotic or chemotherapeutic agents needed to eliminate the suspected causal agents of infection. The authors assessed MIC of 9 selected antibiotics and chemotherapeutic agents (metronidazole, chloramphenicol, clindamycin, azlocillin, mezlocillin, cephoxitin, oxytetracycline, lincomycin, erythromycin) by the dilution method in blood agar in 157 strains of the most important types of the genus Bacteroides which were isolated in 1985-1989 from different clinical materials in Bratislava and in Halle (GDR). All tested strains were sensitive to metronidazole and chloramphenicol. Resistance to clindamycin was very rare. Strains resistant to azlocillin, mezlocillin and cephotoxin were more frequent. A high resistance to lincomycin, oxytetracycline and erythromycin was found. Difference in sensitivity of strains from the CSSR and GDR were slight. Similarly results of the present work differed little from those of previous work.     

Phenotypic and molecular typing of nosocomial methicillin-resistant Staphylococcus aureus strains susceptible to gentamicin isolated in france from 1995 to 1997

Methicillin-resistant strains susceptible to gentamicin (Gm(s) MRSA) have emerged since 1993 in several French hospitals. To study whether particular clones have spread in various French cities and whether some clones are related to gentamicin-resistant (Gm(r)) MRSA strains, various methods (antibiotyping, phage typing, determination of SmaI macrorestriction patterns before and after hybridization with IS256 transposase and aacA-aphD probes) were used to compare 62 Gm(s) MRSA strains isolated from 1995 to 1997 in nine cities and 15 Gm(r) MRSA strains. Eighteen major SmaI genotypes were identified, of which 11 included only Gm(s) MRSA strains and 5 included only Gm(r) MRSA strains. Each of the Gm(r) MRSA strains contained 6 to 13 SmaI fragments hybridizing with the insertion sequence IS256, of which a single band also hybridized with the aacA-aphD gene. No such hybridizing sequences were detected in 60 of the 62 Gm(s) MRSA strains. Thus, the divergence between Gm(r) and Gm(s) MRSA strains is revealed, not only by their distributions in distinct SmaI genotypes but also by the differences in hybridization patterns. Two of the 62 Gm(s) MRSA strains had the uncommon feature of carrying several SmaI bands hybridizing with IS256, suggesting that they are possibly related to the Gm(r) MRSA strains grouped in the same SmaI genotype. Five of the 11 SmaI genotypes including only Gm(s) MRSA strains contained strains from diverse cities, isolated during different years and with different antibiograms, suggesting that some clones have spread beyond their cities of origin and persisted.     

Susceptibilities of Chryseobacterium indologenes and Chryseobacterium meningosepticum to cefepime and cefpirome.

In vitro activities of cefepime and cefpirome against 96 isolates of Chryseobacterium indologenes and 21 of C. meningosepticum were determined by the agar dilution method. Overall, cefepime was more active than cefpirome against C. indologenes (MIC at which 50% of the isolates were inhibited [MIC50] and MIC90, 4 and 16 microg/ml, respectively, for cefepime and 8 and 128 microg/ml, respectively, for cefpirome). Both agents had poor potency against C. meningosepticum (MIC50 and MIC90, 64 and >256 microg/ml, respectively, for cefepime and 128 and >256 microg/ml, respectively, for cefpirome).     

Prospective, randomised, multicentre study of meropenem versus imipenem/cilastatin as empiric monotherapy in severe nosocomial infections

The clinical and bacteriological efficacy and the tolerability of meropenem versus imipenem/cilastatin (both 1 g t.i.d.) in severe nosocomial infections were compared in a multicentre, randomised, nonblinded study. A total of 151 patients were recruited; 133 (66 meropenem, 67 imipenem/cilastatin) were clinically evaluable and 84 (42 meropenem, 42 imipenem/cilastatin) bacteriologically evaluable. Most clinically evaluable patients (90%) were in intensive care units, required mechanical ventilation (72%), and had received previous antibiotic therapy (62%). The mean (±SD) APACHE II score was 15.2 (±6.6) in the meropenem group and 17.8 (±6.8) in the imipenem/cilastatin group. The primary infections were nosocomial lower respiratory tract infections (56% of patients), intra-abdominal infections (15%), septicaemia (21%), skin/skin structure infections (5%), and complicated urinary tract infections (3%); 35% of the patients had two or more infections. There was no significant difference between the meropenem and imipenem/cilastatin groups in the rates of satisfactory clinical (weighted percentage 87% vs. 74%) or bacteriological (weighted percentage 79% vs. 71%) response. There was a slightly higher rate of clinical success with meropenem against primary or secondary lower respiratory tract infection (89% vs. 76%). Drug-related adverse events occurred in 17% and 15% of meropenem and imipenem/cilastatin patients, respectively. Meropenem (1 g t.i.d.) was as efficacious as the same dose of imipenem/cilastatin in this setting, and both drugs were well tolerated.     

Resistance to desiccation and skin fatty acids in outbreak strains of methicillin-resistant Staphylococcus aureus.

Resistance to desiccation and to skin fatty acids was measured in three groups of methicillin-resistant Staphylococcus aureus (MRSA) strains and a group of control strains. Organisms from a large outbreak on a special care baby unit (SCBU), where MRSA had been isolated from staff hands but not from the environment, were significantly more sensitive to drying than strains from a burns unit where extensive environmental contamination had been demonstrated. MRSA from other wards, in the same hospital but not associated with large outbreaks, gave heterogeneous results. Fatty-acid resistance, determined by an agar dilution method, was not associated with strain origin. Some epidemic strains of MRSA were relatively sensitive to desiccation, and the abilities of such strains to spread widely on a SCBU by the hand-borne route could not be explained by enhanced resistance to skin fatty acids.     

Molecular epidemiology of a nosocomial outbreak due to SHV-4-producing strains of Citrobacter diversus.

Over a 6-month period, eight strains of Citrobacter diversus (Citrobacter koseri) resistant to extended-spectrum cephalosporins and monobactams were isolated from seven colonized and/or infected patients from the same intensive care unit. All strains harbored a single large conjugative plasmid which mediated an extended-spectrum beta-lactamase of the SHV-4 type (ceftazidimase phenotype; enzyme pI, 7.8; plasmid DNA hybridization with a blaSHV-specific probe). All strains were characterized by antibiotic resistance pattern analysis, beta-lactamase content analysis, plasmid profiling, ribotyping with EcoRI, and arbitrarily primed (AP)-PCR with primers O8 and O12. Among the eight C. diversus strains, strains Cd5 to Cd12, six isolates (isolates Cd6 to Cd11) were identical by all markers; one strain (strain Cd5) differed by two markers (antibiotype and AP-PCR pattern with primer O8), and the remaining strain (strain Cd12) differed by two other markers (ribotype and AP-PCR pattern with primer O12). Our results suggest that six of the eight SHV-4-producing C. diversus strains studied (strains Cd6 to Cd11) were a single epidemic strain. Strain Cd5 could be related to the epidemic strain; the origin of strain Cd12 remains uncertain.