[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.

Biotyping, ribotyping and esterase electrophoresis as epidemiological tools for Acinetobacter baumannii.

An epidemiological survey was carried out over a one-week period to assess the spread of Acinetobacter baumannii in a medical intensive care unit. Fifty strains were isolated from patients colonized or infected by the organism and from a hospital environment. These strains belonged to biotypes 9 or 18. The rRNA gene restriction patterns (using EcoRI and PvuII as restriction endonucleases) and the esterase electrophoretic profiles were determined on 31 strains, using as comparison strain isolates from another intensive care unit of our hospital and from two other French hospitals. Four EcoRI ribotypes, four PvuII ribotypes and six esterase profiles were identified. All biotype 9 strains isolated in our hospital presented the same ribotype after EcoRI digestion, the same ribotype after PvuII digestion and the same zymotype. The same observation was made on most of the biotype 18 strains. Biotyping is an appropriate method for screening of strains, and ribotyping and esterase electrophoresis could be used as additional methods to delineate outbreaks of nosocomial infections caused by A. baumannii.

Esterase electrophoresis compared with biotyping for epidemiological typing of Acinetobacter baumannii strains.

Forty-nine Acinetobacter baumannii strains belonging to three biotypes and isolated from four hospitals were differentiated by electrophoretic typing of their esterases. Six main kinds of esterases were distinguished by their spectra of hydrolytic activity toward seven synthetic substrates. The electrophoretic variations of these enzymes were used to define ten zymotypes among the three biotypes. Esterase electrophoresis appeared to be more sensitive than biotyping, and could represent an additional marker for epidemiological analysis.

Cleavage of immunoglobulin A1, A2 and G by proteases from clinical isolates of Pasteurella multocida.

Several Pasteurella multocida strains were examined for their ability to produce extracellular enzymes that cleave immunoglobulin A and G (Ig A and Ig G) molecules. Two strains isolated from human pulmonary and genital infections produced proteases that cleaved human IgA and IgG, colostral IgA and human myeloma IgA1 and IgA2. Human IgM was not degraded by these enzymes. Examination of cleavage digests showed two main fragments with different electrophoretic mobilities. The two P. multocida strains produced a protease that cleaved IgA and IgG heavy chains outside the hinge region, and differed in this respect from the hinge-cutting proteases of other bacteria. Protease production may be a virulence mechanism for P. multocida strains.

Dermonecrotic toxin production by strains of Pasteurella multocida isolated from man.

Ninety-four clinical isolates of Pasteurella multocida of human origin were tested for dermonecrotic toxin (DNT) production by three methods: dermonecrotic test in guinea-pigs, Vero cell culture cytotoxicity and ELISA. The strains were isolated from patients living in a rural area with widespread intensive pig breeding. Six strains were found to be toxigenic by the three tests. A major protein band of Mr 145 Kda corresponding to DNT on immunoblots was demonstrated in extracts from these strains. All were isolated from respiratory tract (diseases 5, healthy carriage 1). The difference between isolates from the respiratory tract and isolates from wounds inflicted by pets was statistically significant with regard to DNT production (p less than 0.02). A possible role of the toxin in pulmonary diseases caused by P. multocida has yet to be established.

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.

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 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.

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 salmonellosis in Ille-et-Vilaine in 1983]. / Les salmonelloses humaines en Ille-et-Vilaine en 1983.

224 salmonella strains from human beings, collected by the public and private laboratories of Ille-et-Vilaine during 1983, were serotyped and tested for their susceptibility to antibiotics. Salmonella typhimurium infections were the most frequent, while the other serotypes constituted a small number of strains, generally susceptible to antibiotics. No outbreak due to a multi-resistant serotype was observed. S. typhi and S. paratyphi B represent 6,25% of the strains isolated in this area during the year. Only 22% of the 224 strains collected were resistant to one or more antibiotics.

[Infections and utilization of antibiotics in the hospital, results from a prevalence survey in 1986]. / Infections et usage des antibiotiques à l'hôpital, résultats d'une enquête de prévalence en 1986.

To estimate the rate of infectious diseases and the efficiency of medical care in our hospital, we conducted a prevalence survey of both infections and antibiotic use on 588 inpatients. Results showed that 125 patients (21% of the total) had infections; of these, 56 had nosocomial infections. A third of the patients had been receiving antimicrobial drugs; among them, 30% were receiving the drugs for prophylactic reasons. A misuse in the prophylaxis of infections was determined, particularly in the duration of treatment. The study demonstrated that an effective infection-control program and an antibiotic policy are required in our hospital.

[Identification and rapid determination of the antibiotic resistance of enterobacteria with the automated ABAC identibiogram]. / Identification et antibiogramme rapides des entérobactéries à l'aide du système automatisé ABAC-identibiogramme.

Rapid identification and antimicrobial testing of Enterobacteriaceae by an automated system, the ABAC-Identibiogramme. The system was directly inoculated with a dilution of positive blood culture broths. The results were compared with those of API 20E and the disk method for antibiogram. The ratios of agreement of 94.5% for identification and of 93.4% for antimicrobial testing demonstrate a good accuracy of the ABAC-Identibiogramme.

[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.

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.

[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.

Continuous ambulatory peritoneal dialysis: evaluation of three new culture methods.

Out of 900 dialysates analysed over 15 months at the Rennes Regional Hospital, 116 cloudy specimens with a leucocyte count of more than 100/mm3 were studied. Three methods of laboratory diagnosis based on physical or chemical disruption of leucocytes were evaluated: saponin incorporated in the agar medium, sonication, and the Isolator lysis centrifugation system. Sensitivity was improved (52%) for the three methods combined compared with the standard method (37%). The Isolator system (51%, P < 0.05) gave the best results. Direct microscopic examination nevertheless remains essential for its indicative value (sensitivity 33% of positive dialysates). The leucocyte count was correlated with the percentage of culture positivity (43% from 100-200/mm3 compared with 62% from 500-1000/mm3); 80% of organisms were Gram-positive bacteria and most were coagulase-negative staphylococci. Opportunistic pathogens such as Corynebacterium jeikeium were frequently isolated.

[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.

[Surveillance of nosocomial infections]. / La surveillance des infections nosocomiales.

Exploratory and therapeutic hospital techniques, the frequency of iatrogenic or pathogenic immunodepression and the ever increasing age of the population are factors that augment the risk of nosocomial infections. Some of these infections are unavoidable, but others can be prevented. Appropriate hygienic measures and a rational use of antibiotics contribute to this prevention. We describe here two methods that can be used to watch for nosocomial infections. The one-day recording or prevalence survey method is meant to provide a snapshot image in a given hospital. Longitudinal supervision is more difficult to carry out, but it enables the situation to be more precisely evaluated in each health care centre. Supervision must be regarded as a descriptive stage in the assessment of the local situation as regards the prevalence and incidence of nosocomial infections. It should make it possible to determine priorities in the measures to be taken to prevent most of these infections.