[Suitability of intraoperative autotransfusion]. / Suivi des techniques de récupération de sang en peropératoire.

This work presents the procedure applied by our hospital to assess the quality and security of intra operative autotransfusion. The suitability of the three following variables has to be constantly assessed: performance of the machines to concentrate and wash collected blood, bacterial contamination of processed blood and rate of adverse events. We note that the procedure is applied with participation of medical and nursing staff. Since its setting-up, we note an amelioration of suitable variables.

Characterization of Pasteurella spp. strains isolated from human infections.

This report describes the distribution of species and capsular groups in a collection of 143 strains of Pasteurella recovered from human patients. The organism isolated most frequently was Pasteurella multocida subsp. multocida. As in animals, most of the group A strains were recovered from the respiratory tract. The distribution of species in relation to the animal source suggests that P. multocida subsp. multocida is more infective than other Pasteurella species or subspecies for man.

[Human pasteurellosis: diagnosis, treatment and precautions]. / Le diagnostic, le traitement et la prévention des pasteurelloses humaines.

The circumstances of diagnosis of human pasteurellosis are reviewed. The diagnosis is usually suspected for animal bite or scratch wounds. Conversely, in other infections the diagnosis is only based on bacteriological data. Phenotypic misidentification of Pasteurellaceae from clinical material is common. The phenotypic criteria of identification of the six species of human pathogen Pasteurella are presented. We emphasise that bite wound specimens have to be cultured for aerobic and anaerobic bacteria and yield an average of 5 bacterial isolates per culture. Antibiotic therapy relies upon amino-penicillins or cephalosporins, although b-lactamase producing strains are scarce. Fluoroquinolones can be an alternative for systemic infections. Molecular typing unequivocally points out the risk of transmission from pets to humans. Immunocompromised persons have to be made aware of precautions.

[Antibiotics resistance of Streptococcus pneumoniae. Results from the Brittany registry for 1999]. / Etat de la résistance aux antibiotiques de Streptococcus pneumoniae dans la région Bretagne. Résultats de l'Observatoire Régional du Pneumocoque pour l'année 1999.

Throughout 1999, clinical microbiology laboratories of 13 hospitals in Brittany have recovered Streptococcus pneumoniae isolates in 832 patients, 312 (37.5%) female and 518 (62.2%) male. Two hundred fifty five of them (30.6%) were children. One hundred eighty eight isolates were recovered from blood cultures (22.6%), 16 from CSF (1.9%), 449 from lungs (54%), and 88 from ear exsudates (10.6%).A 5 microgram oxacillin-disk test was used to detect isolates with reduced susceptibility to penicillin G. Determination of MICs of penicillin G, amoxicillin and cefotaxime were then performed by agar dilution method on 402 strains previously categorized resistant or intermediate. Five hundred forty six isolates were PSDP, 33.5% of them were resistant to penicillin G, 2.2% to amoxicillin and 0.2% to cefotaxime. As expected, a decreased susceptibility to beta-lactamins was frequently associated with resistance to macrolides, cotrimoxazole and tetracycline. Among PSDP, the most prevalent serotypes were 23 (23.7%), 14 (23.5%) and 19 (19.1%). In Brittany, the constant rise of PSDP (1993-1994: 28.5%; 1997: 56.4%; 1999: 65.6 %) could be perhaps explain by analysis of social and demographic data.

Susceptibility of Enterobacteriaceae to beta-lactam agents and fluoroquinolones: a 3-year survey in France.

OBJECTIVE: To assess trends in the susceptibility to beta-lactam agents and to fluoroquinolones of clinically relevant Enterobacteriaceae isolated over a 3-year period in 14 French hospital laboratories. METHODS: During the second quarter of 1996, 1997 and 1998, 180 consecutive non-duplicate isolates of Enterobacteriaceae were collected in each center. Sixteen beta-lactams and four quinolones were tested by the disk diffusion method. In addition, the double-disk synergy test was used to screen for the production of extended-spectrum beta-lactamase (ESBL). RESULTS: Totals of 2507, 2312 and 2506 clinical isolates were obtained in each period, respectively. The distribution of Enterobacteriaceae species according to clinical specimens and wards was similar in each study period. No significant variation in the susceptibility rates to beta-lactams and fluoroquinolones was observed, except in Klebsiella pneumoniae and Enterobacter aerogenes. The prevalence of ESBL-producing isolates decreased from 18% to 9% in the former, while it increased from 32% to 54% in the latter. At the same time, the susceptibility to ofloxacin and pefloxacin increased for K. pneumoniae (P < 0.003) and cephalosporinase-producing species (P < 0.05), except Enterobacter spp. CONCLUSION: Over the 3-year study period beta-lactams and fluoroquinolones remained highly active against Enterobacteriaceae clinical isolates, with the exception of E. aerogenes, probably as a result of the dissemination of multiresistant clones in French hospitals.

[Comparative study of the bacteriostatic and bactericidal activity of levofloxacin against Pasteurella strains isolated from man]. / Etude comparative de l'activité bactériostatique et bactéricide de la lévofloxacine vis-à-vis des souches de Pasteurella isolées chez l'homme.

The MICs of seven quinolones, nalidixic acid, pefloxacin, ofloxacin, d-ofloxacin, ciprofloxacin, sparfloxacin and levofloxacin, were determined by agar dilution method comparatively to those of amoxycillin, cefpodoxime, doxycyclin and clarithromycin against 75 clinical isolates of Pasteurella multocida, P. dagmatis and P. canis. Time-kill method was performed for three selected P. multocida isolates. Fluoroquinolones were the most active agents. At concentration of 0.016 mg/L of sparfloxacin or levofloxacin the 75 isolates were inhibited. The MICs of levofloxacin and sparfloxacin showed that the activity of these molecules was two to four times higher than that of the other quinolones studied. Time-kill studies showed a complete killing in six hours with the CMI x 2 of pefloxacin, ofloxacin, ciprofloxacin, sparfloxacin and levofloxacin. This result was obtained more rapidly with the quinolones than with amoxicillin or cefpodoxime. Doxycycline and clarithromycin were devoid of bactericidal activity.

[Investigation of an epidemic of an extended spectrum beta-lactamase producing Escherichia coli in a geriatrics department]. / Investigation d'une épidémie à Escherichia coli à bêta-lactamase à spectre étendu dans un service de gériatrie.

An outbreak of extended-spectrum beta-lactamase-producing (ESBL) Escherichia coli urinary tract infection occurred on one floor of a department of the G. Clemenceau hospital. There were nine cases from March to August 1994, all of which resolved under antimicrobial therapy. Two further cases occurred, leading to a program of routine rectal swab cultures in all patients on the floor involved (first floor). ESBL E. coli carriage was found in six patients. Follow-up rectal swabs were obtained until September 1995 from all patients on the floor who were infected or colonized with ESBL E. coli between March and October 1994. Between March and June 1995, routine rectal swabs were also obtained from the patients on the other two floors of the department. Seven other carriers were detected and also underwent follow-up rectal swabs until September 1995. Ribotyping of the strains done at the national enteric molecular typing center (Pasteur Institute, Paris) demonstrated four patterns (CM1, CM2, CM3, and CM4). The strains from the first six carriers all shared the CM1 pattern, whereas among the seven carriers detected later, four (second and third floors) had a CM1 strain and the remaining three (first floor) each had a strain of the three other ribotypes (CM2, CM3, and CM4). ESBL E. coli can be carried in the digestive tract for several months. During the outbreak, the same strain was found in all the patients, whereas a variety of strains were found after the outbreak in patiens without clinical symptoms due to ESBL E. coli. Routine rectal swab cultures are useful for evaluating the extent of outbreaks. However, the only effective outbreak control measures are compliance with strict hygiene procedures (signalization, technical and geographic isolation) and appropriate antimicrobial therapy. These measures were effective at the G. Clemenceau Hospital since there have been no further cases of ESBL E. coli since they were implemented. The rectal swab program has therefore been stopped, whereas strict universal hygienic precautions are still adhered to faithfully.

Comparative antimicrobial resistance and genomic diversity of Escherichia coli isolated from urinary tract infections in the community and in hospitals.

Well-defined community- and nosocomially-acquired isolates of Escherichia coli responsible for urinary tract infections were studied for their resistance to beta-lactams, quinolones, and co-trimoxazole, antibiotics widely used for treatment of urinary infections. For each strain, an antibiogram was obtained using the Vitek automat, which estimates the minimal inhibitory concentrations of various drugs. Nosocomial strains were significantly more amoxycillin-resistant than community strains (P = 0.01) and were also significantly more resistant to co-trimoxazole (P = 0.025) and first generation quinolones (P = 0.02) than the latter. To determine whether this was due to transmission of strains within the hospital, DNA restriction patterns, established using XbaI enzyme and separation by pulsed-field gel electrophoresis, were compared. Extreme genomic diversity was found among both the community and nosocomial strains. The increased frequency of resistance among nosocomial strains is thus not due to transmission of resistant hospital strains but probably results from the selection of resistant strains from the endogenous flora of patients.

Characterisation of dermonecrotic toxin-producing strains of Pasteurella multocida subsp. multocida isolated from man and swine.

Thirty-six isolates, from man or swine, of Pasteurella multocida subsp. multocida producing (n = 13) or not producing (n = 23) the dermonecrotic toxin (DNT) were studied by numerical analysis, capsular typing and ribotyping. Toxigenic strains were also characterised by restriction fragment length polymorphism (RFLP) of the toxA gene and pulsed-field gel electrophoresis (PFGE). Numerical analysis differentiated the Pasteurella species and subspecies, but did not discriminate between toxigenic and nontoxigenic strains. RFLP demonstrated that toxA was located in a conserved part of the chromosome of all toxigenic strains. Ribotyping provided evidence of a close association between DNT production and one of the six EcoRI ribotypes designated as E2. In contrast, PFGE provided evidence for significant DNA polymorphism amongst the toxigenic strains. Results of phenotypic and genotypic studies suggested that toxigenic strains do not form a clone within the subspecies multocida. No difference was found between toxigenic strains of porcine or human origin by biochemical characterisation, capsular serotyping or genomic typing methods.

Increasing incidence and comparison of nalidixic acid-resistant Salmonella enterica subsp. enterica serotype typhimurium isolates from humans and animals.

We determined the resistance to quinolone of 309 Salmonella enterica subsp. enterica serotype Typhimurium strains isolated from humans and animals (cattle, pigs, or poultry) in 1995 or 1996. Nalidixic acid resistance increased from 8.5% in 1995 to 18.6% in 1996. The highest resistance levels correlated with a mutation at Ser-83 (or Asp-82). All strains remained ciprofloxacin susceptible. Human and animal isolates were compared by pulsed-field gel electrophoresis, and the banding patterns of the human isolates most closely matched those of the bovine isolates.

[Growing incidence of nalidixic acid resistance and sensitivity to quinolones in Salmonella typhimurium strains isolated from man or animal]. / Incidence croissante de la résistance à l'acide nalidixique et sensibilité aux quinolones des souches de Salmonella typhimurium isolées chez l'homme ou l'animal.

To determine the prevalence of quinolone resistance in Salmonella typhimurium strains from humans or animals (cattle, poultry, swine), the S. typhimurium strains isolated at a teaching hospital and at the central veterinary laboratory of the same district between January 1, 1995, and December 31, 1996 were studied. Susceptibility to nalidixic acid was determined using the disk diffusion method. Strains with decreased susceptibility to nalidixic acid were subjected to minimal inhibitory concentration (MIC) determination for pefloxacin, ofloxacin, ciprofloxacin, norfloxacin, levofloxacin, and grepafloxacin. Decreased susceptibility to nalidixic acid was demonstrated for 41 of the 309 strains studied and increased from 8.5% in 1995 to 18.6% in 1996. MIC90 values of fluoroquinolones for strains with decreased susceptibility to nalidixic acid were lower than 1 mg/L, which is the cutoff above which a strain is classified as susceptible, but were higher than for strains that were susceptible to nalidixic acid. These low levels of resistance may be the first step in selection of mutant strains with high levels of resistance to fluoroquinolones. This warrants continued monitoring of resistance of Salmonella to fluoroquinolones.

[Isolation of Escherichia vulneris in drinking water]. / Isolement de Escherichia vulneris dans l'eau d'alimentation.

Over a 2-year period, we performed 33 bacteriological controls of drinking water supplied by refrigerated fountains located in a nursing home. Amongst 24 strains of gram-negative bacilli isolated from 16 samples. 10 were identified as belonging to the species Escherichia vulneris. Viable bacterial counts were always less than 10 ufc/100 ml. During the same period no clinical isolate of E. vulneris was recovered from the nursing home. The signification of E. vulneris in drinking water is unknown. However, considering that E. vulneris has been implicated as cause of various infections, its presence in potable water supply systems would seem to be a potential risk factor for severely immunocompromised patients.

Comparative activities of new fluoroquinolones, alone or in combination with amoxicillin, trimethoprim-sulfamethoxazole, or rifampin, against intracellular Listeria monocytogenes.

We studied the activities of the new fluoroquinolones clinafloxacin, levofloxacin, ofloxacin, and sparfloxacin alone or in combination on the intracellular growth of Listeria monocytogenes. Against intracellular growth of the four strains tested, a similar reduction of the bacterial count was obtained with clinafloxacin at the dose of 10 x MIC (delta log10 CFU/ml = -2.19 +/- 0.24), with levofloxacin at 8 x MIC (delta log10 CFU/ml = -2.28 +/- 0.25), and with sparfloxacin at 4 x MIC (delta log10 CFU/ml = -2.16 +/- 0.21) after 24 h of incubation. The combination of the quinolones with trimethoprim-sulfamethoxazole or amoxicillin did not show a substantial increase in activity compared to the fluoroquinolone alone. Antagonism with rifampin was strongly suggested. No modification of the MIC was observed after 20 successive infections of HeLa cells and contact with subinhibitory concentrations of clinafloxacin, levofloxacin, and sparfloxacin for 24 h. We conclude that clinafloxacin, levofloxacin, or sparfloxacin could represent a therapeutic alternative to amoxicillin for the treatment of Listeria infections in adults, especially clinafloxacin, whose MIC is low (0.06 to 0.12 micrograms/ml), and whose best activity against intracellular L. monocytogenes was obtained at a concentration of 1.2 micrograms/ml, which is similar to clinically achievable levels. The results must be confirmed in an experimental model.

First European case of gastroenteritis and bacteremia due to Vibrio hollisae.

Vibrio hollisae is a pathogenic Vibrio species known to cause gastroenteritis in humans after the consumption of shellfish. All cases of infection reported previously were restricted to the Atlantic and Pacific coasts of the United States. A case of gastroenteritis and bacteremia in a previously healthy 76-year-old man who ate cockles from the Quiberon Bay in Brittany, France, is described. This is the first report of Vibrio hollisae infection in Europe.

Epidemiologic analysis of glycopeptide-resistant Enterococcus strains in neutropenic patients receiving prolonged vancomycin administration.

Vancomycin-resistant enterococci have been isolated with increasing frequency since 1988. Thus far, most of these resistant enterococci have belonged to the Enterococcus faecium species, and epidemiological studies have shown a wide diversity among interhospital and intrahospital isolates. This report presents an epidemiologic investigation of 25 vancomycin-resistant Enterococcus strains--24 Enterococcus faecium and one Enterococcus gallinarum--isolated from the stools or blood of adult patients receiving intravenous vancomycin prophylaxis during neutropenia and hospitalized in a single hematologic unit. Macrorestriction patterns of total DNA and of ribosomal DNA regions were used to analyze the strains. Strains produced different total DNA restriction fragment length polymorphism patterns after SmaI digestion. Ribotyping was less discriminative than pulsed-field gel electrophoresis. The results confirmed the genetic unrelatedness of the strains. Prolonged vancomycin administration, commonly used in hematologic units, could be involved in the selection of endogenous resistant enterococcal strains.

[Susceptibility to quinolones of Salmonella isolated from men and animals]. / Sensibilité aux quinolones des Salmonella isolées chez l'homme et l'animal.

Fluoroquinolones are efficacious antibiotics for the treatment of Salmonella infections in humans. One of these quinolones, enrofloxacin, a precursor of ciprofloxacin, is used to treat respiratory infections in calves and poultry. There is a risk of developing resistant strains of Salmonella in animals, which may then contaminate humans. To evaluate current susceptibilities of Salmonella strains to quinolones, we collected 95 strains belonging to various serotypes in a district of intensive breeding (the Ille-et-Vilaine Département): 54 human strains, 24 bovine strains, and 17 poultry strains. The technique of dilutions in agar medium was used to determine Minimal Inhibitory Concentrations (MICs) for the following antibiotics: nalidixic acid, norfloxacin, pefloxacin, ofloxacin, ciprofloxacin, levofloxacin and clinafloxacin. Our results showed that human Salmonella strains remained very susceptible to quinolones. Only 3 animal strains had nalidixic-acid MICs > 128 mg/l. For these 3 strains, pefloxacin and ciprofloxacin MICs were respectively 1 or 2 mg/l and 0.25 mg/l. For all the other human and animal strains the nalidixic-acid MICs were < 4 mg/l, and their MICs 90 were: 0.12 mg/l for ofloxacin and norfloxacin, 0.06 mg/l for pefloxacin, 0.03 mg/l for ciprofloxacin, 0.016 mg/l for levofloxacin, and 0.004 mg/l for clinafloxacin.

[Pasteurelloses]. / Les pasteurelloses.

According to the genetic relationships among Gram-negative bacilli the genus Pasteurella is included with the genus Haemophilus and the genus Acinobacillus within the family Pasteurellacae. Pasteurella multocida, the type species, is responsible for the majority of human Pasteurella infections. P. multocida is a member of the normal flora in the upper respiratory tract of many mammals or birds. It causes sporadic or epidemic diseases among different animal species, particularly pneumonia and atrophic rhinitis in swine in intensive breeding stations. The most common human infection with P. multocida is a local cellulitis following dog or cat bites and scratches. Serious local complications are sometimes responsible for prolonged disability. The respiratory tract is the second human source of P. multocida isolates. The frequency of recovery of P. multocida from oropharynx of apparently healthy pig breeders suggests that respiratory pasteurellosis could be an occupational disease. The mechanisms of virulence of P. multocida are unclear. Several factors are involved: capsules preventing phagocytosis, a dermonecrotic toxin causing experimental atrophic rhinitis, hyaluronidase, neuraminidase and proteases. Penicillin is considered to be the drug of choice for Pasteurella infection. Tetracyclin is efficient for bites but has no bactericidal effect. Oxacillin, first-generation cephalosporins, macrolides and aminoglycosides have poor activities. In the case of beta-lactamase producing strains a bactericidal effect could be achieved with fluoroquinolones or third generation cephalosporins.

[Bactericidal activity of ciprofloxacin and sparfloxacin. Comparison with others active antibiotics against Pasteurella multocida]. / Activité bactéricide de la ciprofloxacine et de la sparfloxacine. Comparaison avec celle des autres antibiotiques actifs sur Pasteurella multocida.

Bactericidal activities of ciprofloxacin and sparfloxacin were compared to other antibiotics active against human isolates of Pasteurella multocida. Three human isolates of Pasteurella multocida were used for killing-curve studies with ciprofloxacin and sparfloxacin comparatively to others antibiotics. At 2x the MIC, ciprofloxacin and sparfloxacin exhibited a killing of more than 99.9% of the initial viable cells that was achieved within 6 h of incubation. These activities were faster than those of amoxycillin and cefpodoxime. No regrowth was observed after 24 h of incubation. Doxycycline and clarithromycin used at MICx2 had no bactericidal activities. It was concluded that fluoroquinolones, namely ciprofloxacin and sparfloxacin, can be considered having good bactericidal activity against P. multocida.

Clinical evaluation of the Vital system compared with the Hémoline diphasic method for the detection of aerobic blood cultures.

In a collaborative study at two university hospitals, the recovery of microorganisms and the speed of detection of microbial aerobic growth by the Vital system (bioMérieux) and a diphasic conventional blood culture system were compared. The Vital system monitors each blood culture bottle every 15 minutes to detect inactivation of fluorescent suspended in the broth medium due to microbial growth. A total of 1086 comparisons were made between the two systems, yielding a total of 117 isolates. Microorganisms were recovered more often from the Vital aerobic bottles (p < 0.05). The Vital system detected 43% of the microorganisms within the first 12 hours of incubation whereas in the same time the conventional system detected only 5% of the microorganisms. The results demonstrate that the Vital system is a reliable, continuous monitoring, fully automated system and an attractive alternative to conventional blood culture methods.