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Laboratory waste management must ensure the safety of patients and staff, limiting the environmental impacts and control waste disposal budget. Sorting of waste must be carried out at the source. The packaging must be adapted, allowing easy identification of specific disposal routes. With regard to wastes for human or animal health care and/or related research (DASRI), packages must comply with the regulations, standards and ADR if necessary. Storage provisions differ according to the amount of DASRI produced. Waste collection is carried out directly on the place of activity by a certified service provider. Non pre-treated DASRI is incinerated in specific approved plants for a T ° > 1,200 °C. Special provisions also exist for chemical waste and radioactive waste, the latter being regulated by ANDRA.

Risk management in Medical Biology Laboratory (MBL) which includes hygiene and waste management, is an integrated process to the whole MBL organisation. It is composed of three stages: risks factors identification, grading and prioritization, and their evaluation in the system. From the legislation and NF EN ISO 15189 standard's requirements viewpoint, prevention and protection actions to implement are described, at premises level, but also at work station environment's one (human resources and equipments) towards biological, chemical, linked to gas, to ionizing or non ionizing radiations and fire riks, in order not to compromise patients safety, employees safety, and quality results. Then, although NF EN 15189 standard only enacts requirements in terms of prevention, curative actions after established blood or chemical exposure accident are defined.

The ability to form a biofilm seems to play an essential role in the virulence of coagulase-negative staphylococci (CoNS) by permitting them to cause persistent prosthetic device-related infections. The most clearly characterized component of staphylococcal biofilms is the polysaccharide intercellular adhesin (PIA) encoded by the icaADBC operon. In the present paper, we assess the link between the ability to form a biofilm (Bf+/-), to synthesize PIA (PIA+/-) and the presence of the ica locus (ica+/-). For this purpose, 66 CoNS strains were tested in vitro. Seventy three percent of all strains revealed presence of the ica locus (ica+), and therefore were potentially able to produce PIA and to form a biofilm. However, the characteristics observed indicated that 15% of all strains were biofilm forming without PIA production (Bf+, PIA-, ica+/-) while 8% were PIA producers without biofilm formation (Bf-, PIA+, ica+). On the basis of the obtained data we conclude that (i) PIA synthesis alone is not sufficient to produce a biofilm and (ii) staphylococci can also form a biofilm without producing PIA.

The intrinsic resistance of the Mycobacterium tuberculosis complex (MTC) to most antibiotics, including macrolides, is generally attributed to the low permeability of the mycobacterial cell wall. However, nontuberculous mycobacteria (NTM) are much more sensitive to macrolides than members of the MTC. A search for macrolide resistance determinants within the genome of M. tuberculosis revealed the presence of a sequence encoding a putative rRNA methyltransferase. The deduced protein is similar to Erm methyltransferases, which confer macrolide-lincosamide-streptogramin (MLS) resistance by methylation of 23S rRNA, and was named ErmMT. The corresponding gene, ermMT (erm37), is present in all members of the MTC but is absent in NTM species. Part of ermMT is deleted in some vaccine strains of Mycobacterium bovis BCG, such as the Pasteur strain, which lack the RD2 region. The Pasteur strain was susceptible to MLS antibiotics, whereas MTC species harboring the RD2 region were resistant to them. The expression of ermMT in the macrolide-sensitive Mycobacterium smegmatis and BCG Pasteur conferred MLS resistance. The resistance patterns and ribosomal affinity for erythromycin of Mycobacterium host strains expressing ermMT, srmA (monomethyltransferase from Streptomyces ambofaciens), and ermE (dimethyltransferase from Saccharopolyspora erythraea) were compared, and the ones conferred by ErmMT were similar to those conferred by SrmA, corresponding to the MLS type I phenotype. These results suggest that ermMT plays a major role in the intrinsic macrolide resistance of members of the MTC and could be the first example of a gene conferring resistance by target modification in mycobacteria.

Between January 1997 and April 2002, 73 consecutive invasive strains of Streptococcus pneumoniae were isolated from children under 16 years of age in four hospitals in suburban Paris. Their genetic diversity was investigated by serotyping and analysis of pulsed-field gel electrophoresis restriction patterns. Antibiotic susceptibility patterns were analysed by disk susceptibility testing and determination of minimal inhibitory concentrations. The genetic basis of macrolide resistance was investigated by polymerase chain reaction. Studies of penicillin and vancomycin tolerance were performed for each strain. Despite the high prevalence (45.2%) of penicillin-nonsusceptible Streptococcus pneumoniae, resistance to amoxicillin (1.4%) was rare, and no strain was resistant to cefotaxime. Overall, 4.1% of pneumococcal strains were resistant to penicillin. Penicillin or vancomycin tolerance was not detected in any of the 73 strains studied. Of the erythromycin-resistant strains (48%), all but one carried the ermB gene. No strains showing a decreased susceptibility to ciprofloxacin (MIC, >4 mg/l) or overexpressing an efflux pump inhibited by reserpine were isolated. The serotypes found, in order of frequency, were as follows: 18C, 14, 6B, 19F, 19A, 9V, 23F, 1, 7F, 9A, 38. Strains of penicillin-nonsusceptible Streptococcus pneumoniae belonged predominantly to serotypes 14, 6B, 9V, 9A, 23F, 19F and 19A. The seven-valent conjugated vaccine covered 85.5% of the serogroups isolated in children under 2 years of age and 65.6% of the serogroups identified in children over 2 years of age. The genetic analysis showed a high identity for some serotypes, such as 14/9V, 6B and 23F. The use of the seven-valent conjugated vaccine is a critical measure to prevent invasive pneumococci infections in children in the Ille de France area.

The E-test is convenient for testing susceptibility of anaerobes. From September 1998 to September 1999, 194 strains (105 Gram-positive bacteria, 89 Gram-negative bacteria) of clinically relevant samples were tested against five antibiotics benzylpenicillin, amoxicillin-clavulanic acid, clindamycin, metronidazole and imipenem on blood agar plates. Resistance to benzyl penicillin is widespread and Gram-negative bacteria and resistance to amoxicillin-clavulanic acid is exceptional. Metronidazole is very effective against anaerobes except non-spore-forming aerotolerant Gram-positive rods and Peptostreptococcus micros.

Unlabelled: Few data are available about pediatric imported malaria, whereas the number of cases seems in constant increase., Patients and Methods: all pediatric malaria cases diagnosed by a positive thin or thick blood film at the Versailles Hospital, from January 1997 to December 2001, were studied retrospectively., Results: sixty cases of pediatric imported malaria were studied. They were 58 cases of uncomplicated malaria and 2 cases of severe malaria; 85% of the children had travelled to sub-Saharan Africa and 15% to Oceania; 90% of the children were of African origin. Plasmodium falciparum was found alone in 84% of the cases. The anti-malarial chemoprophylaxis was inappropriate in 92% of the cases. No child had profited from preventive measures against mosquitos. Fever > 37,5 degrees C was observed in 100% of the cases. The other clinical signs were present in less than 50% of the cases. The median of haemoglobin and platelet was 10.5 g/dL and 141,000/mm(3), respectively. After treatment, the evolution was good in all the cases, without relapse or any consequences., Discussion/conclusion: our study, in agreement with the national data, confirms the increase in the number of case of pediatric imported malaria, and underlines the mediocrity of the prevention, in particular in term of anti-malarial chemo-prophylaxis. These data, in a context of regular increase of international travels to endemic areas, suggest the necessity to improve the information of the general public, and the urgency of a better staff training of health care workers concerning malaria, in order to improve the prevention and the treatment of this potentially fatal disease.

Using 106 clinical isolates of mycobacteria, we showed that INNO-LIPA Mycobacteria assay is an excellent tool to rapidly identify the most frequently isolated nontuberculous mycobacteria, in one procedure. It may be used as an additional technique to AccuProbe assay, which remains the fastest and the cheapest tool for a rapid and accurate identification of the M. tuberculosis complex.

Objectives: Group B Streptococcus (GBS) continues to be the most important bacterial cause of sepsis, meningitidis in newborns. American consensus guidelines have been published. They recommended the use of either risk-based strategy or screening-based approach for GBS colonization in pregnant women to identify candidates for intrapartum prophylaxis. Screening consists of obtaining vaginal and anorectal specimens for culture at 35 to 37 weeks' gestation. The aim of this prospective study was to assess the usefulness of systematic and concomitant GBS screening by rectal and vaginal swab in pregnant women. A questionnaire designed to determine the risk factors for colonization by GBS was completed., Materials and Methods: We have screened 370 pregnant women with rectal and vaginal swab., Results: Fifty seven (15.4%) women had positive GBS cultures. Of those women, the rectum and the vagina were the only site of colonization in 16 (4.3%) and 8 (2.2%) women respectively. None of the factors studied was significantly associated with GBS colonization., Conclusion: Detection of GBS is enhanced by 40% by using vaginal and anorectal swabs compared to a vaginal swab alone. No studied factor appeared to predict GBS colonization, which incited us to screen all pregnant women.

In deep seated candidiasis, only 40% of blood cultures are positive. The aim of the study was to investigate circulating Candida albicans mannan and anti-mannan antibodies as a possible help for the diagnosis of deep seated candidiasis. We have compared the results to the detection of IgM by Elisa and antibodies by immunoflourescence. The best tests, in accord to their sensitivity and specificity, are the mannan antigenemia (43% and 100%) and IgM (86% and 100%) and have to be used together.

Objectives: The resurgence of whooping cough observed in France convinced us to develop a specific PCR assay to detect B. pertussis in nasopharyngeal secretions in parallel of the culture. The aim of our study was to show the value of the PCR in routine diagnosis., Material and Methods: From November 1996 to August 2000, in two hospitals located in the Yvelines (France), the children consulting for a cough compatible with the diagnosis of whooping cough were included in this study. A questionnaire including clinical, biological and radiological items was completed for each one of these patients. A culture of Bordetella and a detection by PCR of B. pertussis were carried out on each nasopharyngeal aspirate. The diagnosis of whooping cough was retained if the detection was positive in PCR and/or culture., Results: Among the 215 investigated children with suspected cases of whooping cough, the diagnosis was positive for 45 (20.9%), of which 39 were less than one year old (median: three months). Sixteen (35.5,%) were positive at the same time for both the PCR and the culture, 26 (57.8%) for only PCR and three (6.7%) for only culture. The PCR was positive in 93.3% of the cases. The results were obtained with an average time of 48 hours. The culture was positive in 61.2% of the cases with an average time of six days. The monthly distribution of the cases of whooping cough was very inhomogeneous and of epidemic appearance. The majority of the cases was located between two periods: 42% between November 1996 and September 1997 and 40% between November 1999 and August 2000. Among the infected children, 15 were less than two months old and were not yet vaccinated; among the 24 others infants, a delay in the vaccine calendar was noted in 50% of the cases. Four children between six and 14 years old were correctly vaccinated. The evolution was favourable in all the children., Conclusion: The PCR due to its sensitivity, its specificity and its rapidity offers to the clinician a powerful tool for the diagnosis of whooping cough. Nevertheless, the culture must be associated with the PCR, in order to follow the epidemiology and the sensitivity to antibiotics of B. pertussis.

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.

The newly developed Hemofast MRSA system for Staphylococcus aureus identification and methicillin-resistant staphylococci (MRS) detection in blood culture broths was evaluated in 106 Bactec broths containing grapelike clusters of Gram-positive cocci. All 26 S. aureus positive broths were correctly identified by Hemofast MRSA within two hours, and 81% were identified within one hour. Sensitivity and specificity were both 100%. Accuracy of MRS detection was 100%, i.e., no discrepancies versus the agar diffusion method were found. When the 28 broths containing coagulase negative staphylococci (CNS) were tested using an inoculum ten fold larger than for S. aureus testing, a number of discrepancy were recorded. For 49 other broths containing CNS, accuracy was 98.5% when the test was interpreted based on the growth rate of the strain, according to the manufacturer's instructions. One broth containing S. epidermidis strain susceptible yielded a false result with Hemofast MRSA, indicating that it probably contained a contaminant. The other discrepancies occurred with specimens containing mixed populations with CNS or a Micrococcus strain. Most (85%) results were available within 4 hrs 30 min, irrespective of the S. aureus or CNS strains. Avoiding the isolation step on agar, Hemofast MRSA saves 24 to 48 hours, thus allowing earlier antistaphylococcal treatment.

Staphylococcus aureus identification is one of the priorities of the microbiological diagnosis of the staphylococcal infections. Current identification methods are carried out after a first step of colony isolation on agar media. We describe a fluorogenic method for S. aureus identification, which is directly applied to blood culture broths. This method uses a gel tube which allows an optimized microculture of the bacteria. 129 clinical samples of blood cultures (HEC) containing gram positive cocci in grapelike clusters (35 Vital bottles, 94 Bactec bottles), and 77 inoculated blood culture (HE) with collection strains of S. aureus are included in the study. Bacteria are concentrated and separated from other components of sample in the gel tube. Staphylococci are revealed during a microculture in the gel phase, by using a colorimetric substrate of their dehydrogenases. Then, staphylococci are recovered in an adapted culture medium containing human prothrombin and a fluorogenic substrate, which is specific for the staphylocoagulase. After 1 to 2 h incubation at 37 degrees C, a blue fluorescence shows the presence of S. aureus. Among the 40 HEC containing S. aureus the test is positive for 37 samples. For 3 cases, the test is not interpretable, due to non lysis red blood cells in the gel phase of the tube. No false positive result is observed for the HEC containing coagulase-negative staphylococci. Moreover, our method is positive with the 77 HE. 94.7% of tested samples (HEC and HE) show a fluorescence after only one hour and half. Sensibility and specificity are both 100%. We propose a rapid method for S. aureus identification directly applied to blood culture broths. This method saves 24 h, avoiding the isolation step on agar. Therefore, the treatment of staphylococcal infections and possible isolation measures could earlier set up.

Staphylococcus aureus is responsible for septicaemia and serious nosocomial infections. A rapid and specific identification of this species is of great importance in clinical microbiology. Current methods for S. aureus identification require a 18 to 24 h-incubation. We describe a two hour-identification method based on the detection of the staphylocoagulase, using human prothrombin and a chromogenic substrate. 242 staphylococcal strains (160 S. aureus, 82 coagulase-negative staphylococci (CNS)) were collected from 4 French hospitals. They have been identified by the following methods: (i) clotting of citrated rabbit plasma, which is considered as reference method; (ii) biochemical tests (Rapidec Staph and Api Staph or ID 32 Staph); (iii) and agglutination test (Pastorex Staph or Pastorex Staph-plus). A strain of S. intermedius was provided by the Collection of the Pasteur Institute (Paris). An adapted culture medium is inoculated with staphylococci and adjusted to 2 Mac Farland unities. This medium is then mixed to an equal volume with a human prothrombin solution and the chromogenic substrate. After 1 to 2 hours incubation at 37 degrees C, the strength of the yellow colour of the mixture is observed to the naked eye, or measured at 405 nm with a spectrophotometer. Fifteen chromogenic tripeptides having a thrombin-like affinity and paranitroanilin as leaving group were compared. With the substrate which has the higher hydrolysis velocity and enzymatic affinity (SQ149), all S. aureus strains gave a positive result: 94.7% of the methicillin-susceptible S. aureus were detected after 1 hour incubation, but only 52.3% of the methicillin-resistant S. aureus. 98.4% of the methicillin-resistant S. aureus were detected after 2 hours. No false positive result was observed for the 82 CNS strains. The chromogenic method shows good within-run and day-to-day precision tests. It doesn't need any complementary test. The sensitivity and the specificity are 99.4% and 100% respectively.

A. baumannii is a multiresistant bacteria which is recognised as responsible for nosocomial infections and hospital outbreaks. The control of these outbreaks depends on the strain's typing and on the fight's policy against nosocomial infections. An outbreak of A. baumannii is occurred to patients who were hospitalized in Centre Hospitalier de Versailles. To investigate this outbreak, we have determined the biotype (Bouvet's method), the succeptibility pattern (disk diffusion and agar dilution results were analysed with the hierarchical classification and main component analysis) and the total DNA macrorestriction pattern (Pulse Field Gel Electrophoresis using SmaI restriction enzyme). A risk factors for A. baumannii acquisition were delineated in case-control study. During 2 years, 38 patients have been infected or colonized to A. baumannii. Thirty two patients were hospitalized in ICU. We studied 38 non repetitive clinical isolates and 9 strains of the patient's rooms. Four biotypes were defined by the Bouvet's typing method. Fourteen groups were obtained when succeptibility results were analysed with the hierarchical classification and 6 with the main composant analysis. The molecular typing permit us to define 4 epidemic and 6 sporadic strains. All the epidemic strains were isolated on ICU hospitalized patients. Our study has shown wide contamination in patient's rooms (Water tap, dry surfaces, patient's mattresses...). Environmental objects have been a major risk factor for A. baumannii acquisition. The control of this outbreak has been possible by application of hygienic measures (hands washing, isolment, meticulous cleaning of the ICU and environmental controls). No new case is occurred in the last year. Typing methods and case-control study are necessary to investigate cross-infections and take efficient measures against these outbreaks.

Twenty-one strains of Torulopsis glabrata have been isolated in the hospital of Versailles during a three months period, in eight patients, and related to nosocomial infections. In order to know if they have been cross infected, a genotypic analysis (pulsed field electrophoresis) has been performed. The strains from four patients had the same karyotype, suggesting they had probably crossed infections. Since this kind of analysis is expensive and time consuming, we have tried a more practical approach. The susceptibility of the strains has been tested by Etest for five antifungal agents: amphotericin B, 5-fluorocytosine, ketoconazole, fluconazole et itraconazole. Statistical analysis of these results has not given the same classification of the strains of T. glabrata. Karyotyping by pulsed field electrophoresis of strains seems to be a better method to confirm strain clonality.

During 2 months (01.01.95-28.02.95), we noted for each Staphylococcus aureus (SA) isolates, identification and susceptibility methods, hospital size, medical speciality, source, penicillin (P) and methicillin (M) susceptibility. For each non repetitive methicillin resistant SA we noted phenotype of aminoglycoside resistance of SA to 6 antistaphylococci drugs. We isolated 399 SA, SAP sensitives 6.5%, methicillin resistant SA (MRSA) 31.8%. MRSA in acute care unit 28.4%, non acute care unit 61.7%, surgery 28.6%, medicine 36.5%, orthopedics 34.5%, pédiatrics 4%, ICU 38.8%, others 10.3%. MRSA from wound 28.2%, respiratory tract 38.3% blood 22.5%, urine 58.5%, others 25.8% - MRSA kanamycin 1.7%, kanamycin tobramycin 36.2%, kanamycin, tobramycin, gentamicin 56.9% - MRSA erythromycin 79.5%, pefloxacin 91.3%, rifampicin 53.5%, acide fusidique 8.7%, fosfomycine 18.9%, TMP SMZ 5.5%. The proportion MRSA is equivalent among medical specialities predominant in urine cultures and respiratory tract. It should be better to carry on the survey for more than two months in each hospital.

Interferon alpha (INF) used for chronic hepatitis C treatment induces a response in less than 25% of patients. The aim of this study was to appreciate the predictive value of the delay of clearance of hepatitis C virus (HCV) viremia after onset therapy and to compare it with other virological markers such as viral load before treatment and viral type. Thirty one patients with chronic hepatitis C, treated with 3 MU Interferon, 3 times a week for 6 months, were followed until 6 months post-treatment. Response was defined according to the normalisation of transaminases levels and loss of HCV viremia. Five patients were long term responder, eleven patients were complete to relapse responder and fifteen patients were non responder. Serum HCV RNA level, HCV type and serial detection of serum HCV RNA were determined and correlated with the long term response to INF. Patients with long term response had lower pre-INF viral load compared to the complete to relapse responder or to the non responder (p < 0.001). A rapid clearance of serum HCV RNA (1 month) is observed for all the long term responder (p < 0.001). The lowest viral load is also observed in these patients (p < 0 05). By contrast, although the number of patient is low, we were not able to observe a relation between the viral type and the response to treatment. In conclusion this data indicate that the delay of clearance of HCV RNA is also a good predictor of response to INF therapy. Furthermore a rapid clearance of HCV RNA in patients with a very weak pre-INF viral load is strongly associated with long term response (positive predictive value: 100%).

Systemic fungal infections are an important cause of morbidity and mortality among immunocompromised patients. New antifungal agents, such as triazoles, are now available, and the place of in vitro tests has to be discussed. It has been shown that interlaboratory reproducibility of in vitro susceptibility tests against fungi was low, due to the lack of standardization. Recently, the NCCLS defined conditions permitting a good interlaboratory reproducibility. However, the predictive value of in vitro susceptibility tests on the therapeutic outcome remain to be demonstrated, and is now under investigation. At the present time, susceptibility testing can be useful: in patients treated by amphotericin B for a severe fungal infection and who do not improve under therapy; to detect resistance to 5-fluorocytosine; to compare the sensitivity to triazoles before and after treatment, in case of therapeutic failure. Serum levels monitoring is useful to prevent the toxicity due to 5-fluorocytosine and to control the digestive absorption of triazoles, especially the lipophilic compound itraconazole.

A comparative study of two air-bacteria counting machines, MK II (Casella, London) and R.B. (Joubert, Lyon) was carried out. Pre-operation air samples from operating rooms were drawn by the two machines working simultaneously. The results from the two machines are similar, however the RB machine is the more sensitive of the two machines. Moreover, an old operating room without filtrated air had a significantly higher air bacteria contamination (m = 199 +/- 173 bacteria/m3) than a new one with filtrated air (m = 40 +/- 36 bacteria/m3). Samples from laminar air flow (m = 1,3 +/- 1,5 bacteria/m3) were examined as test studies. Working methods and results of air bacteria counts are studied.

Qualitative bacterial samples were taken from 53 appendices which were purulent, or had lymphoid hyperplasia or were normal. The results did not demonstrate a specific bacterial flora responsible for appendicular disease in childhood.

A distal bronchial brushing was performed in 25 children having severe lung diseases. The age of the patients ranged from 7 days to 14 years. Seventeen children were immuno-depressed. Bacteria were isolated five times (2 Staphylococcus aureus, 1 Streptococcus pneumoniae, 1 Serratia marcenscens, 1 Acinetobacter lwoffi), viruses twice (cytomegalovirus, syncytial respiratory virus), Pneumocystis carinii, four times. Cytologic study revealed malignant cells, and cells with intranuclear inclusions, evoking a viral infection. This procedure appears to be harmless in children over 2 years of age. In younger children there is a risk of pneumothorax. the principal indication for distal bronchial brushing is lung disease in immuno-depressed children. It may, however, also be useful in prolonged, treatment-resistant, severe lung diseases.

Cytosolic superoxide dismutase (SOD) activity was found to increase with time during HeLa cell culture, this increase being due exclusively to Mn-SOD. Infection of the cells by Chlamydia trachomatis resulted in a further enhancement of this Mn-SOD activity, whereas cytosolic catalase activity was decreased in these infected cells. Superoxide (O-2.) being able to induce Mn-SOD and to inhibit catalase, these data suggest that Chlamydia trachomatis infection could be responsible for an increase in O-2. production by the infected HeLa cells.

A comparative study of two air-bacteria counting machines, MK II (Casella, London) and R.B. (Joubert, Lyon) was carried out. Pre-operation air samples from operating rooms were drawn by the two machines working simultaneously. The results from the two machines are similar, however the RB machine is more sensitive of the two machines. Moreover, an old operating room without filtrated air had a significantly higher air bacteria contamination (m = 199 +/- 173 bacteria/m3) than a new one with filtrated air (m = 40 +/- 36 bacteria/m3). Samples from laminar air flow (m = 1,3 +/- 1,5 bacteria/m3) were examined as test studies. Working methods and results of air bacteria counts are studied.

The authors report a case presenting with a very rare localization of brucellosis: the isolated involvement of a single bone of the metatarsus, in a child in otherwise excellent general condition. The lesions of the diaphysis and epiphyses induced a fragility of the epiphyseal cartilage and a minimal traumatism resulted in a detachment of the epiphysis which was the first clinical sign. The bacteriological examination of a sample of pus led to the early diagnosis of brucellosis, therefore allowing efficient treatment.

1,707 non replicate clinical strains of Pseudomonas aeruginosa from non teaching hospitals were investigated. Beta-lactam antibiotics were tested by agar disk diffusion method. Variance analysis revealed a significant interaction between strains and experimental laboratories. The bacterial population was grouped according to the diameters of inhibition zones of antibiogram, into five phenotypes. Pseudomonas aeruginosa is frequently isolated from urines and pulmonary sampling, rarely from blood cultures. Resistance to beta-lactams increased significantly with the duration of hospitalisation, and age of patients. Clinical strains are significantly more resistant in blood cultures, and for strains isolated from reanimation. 84% of strains are susceptible to ticarcillin.

The bacterial flora of the small bowel was studied in infants with stasis after surgery. Samples were obtained, with radiological assistance, by duodenal aspiration and then immediately analysed by the laboratory in blind tests, both qualitatively and quantitatively. There were two groups of patients: one receiving parenteral nutrition and a second on enteral elementary diet; patients without any gastro-intestinal tract disease formed the control group. Pseudomonas aeruginosa and Enterobacteria were isolated significantly more often in both groups of patients compared to the control group. Quantitatively, the predominant flora -- and not total flora -- was different in the three groups. When considering bacteriological species, only P. aeruginosa (greater than 10(6)/ml) differed significantly in numbers in both groups compared to the control group. With these bacteriological investigations, the "contaminated proximal small bowel syndrome" is better defined and a group of high-risk patients may be identified.

The conventional serum-dilution bactericidal test used for monitoring antibiotic therapy in severely infected in patients requires 72 h. Use of an automaton would be expected to provide faster results. We tested the MS-2 diagnostic automaton which performs kinetic analysis of bacterial growth. A relation between exponential growth onset time and initial bacterial concentration of the inoculum was first determined using successive ratio 10 dilutions. Sixty-two serum samples from twenty-one patients with endocarditis or septicemia were examined. A selection of findings is presented. Results of the automatic method and serum-dilution bactericidal test are studied comparatively. From our data we define the major advantages of automation: only 0.65 ml of serum and 10 minutes are needed to perform the test and results are obtained in five to twelve hours according to the bacterial strain. This new technique could be integrated in the monitoring protocol of severe infections.

Authors report on a series of ten ventricular-cardiac or peritoneal derivations performed under operative field isolation, air-tight and sterile enclosure that makes possible to perform surgery under strict aspsis. The first results compared with those mentioned in the medical literature, let foresee a fall in the occurence of post-operative infections. The technique is adaptable to neurosurgery and does not complicate the operative process.

A strain of Lactobacillus casei from a pharmaceutical preparation became established in the digestive tract of an axenic child. It was eliminated by the first bacteria occurring after inoculation of the child's digestive tract with a human flora, before the child was taken out of the isolator. Using "gnotoxenic" mice, we found that the elimination of L. casei was due to a bactericidal effect of the Escherichia coli of human origin, within the digestive tract.

Penicillinase producing Neisseria gonorrhoeae PPNG, had been first isolated in France in 1979. Since, they regularly increased if we considered France on the whole. From 1979 to 1986, 284 strains had been collected by a multicentric group. The frequency of isolation was strongly different in France, unknown in some region they rose 12% in specific areas in Paris. The PPNG strains were more frequently isolated from male than female (sex ratio was higher with PPNG than for non producing strain). They were more often responsible of complicated infections in female than male at the same rate than the non producing strains. Auxotype distribution was different between producing and non producing strains. Plasmidic content from african type (Af) was almost the same than from asian type (As). Strains with Af type associated with the conjugative plasmid were increasing.

A technique not involving surgery is described for obtaining axenic (germ-free) newborn animals and human infants by decontamination immediately after birth. Three steps are involved: cleansing ther perineal region of the mother with an iodinated bactericidal solution, washing the newborn with the same solution, and after the newborn has been placed in a sterile isolater, administering a single oral dose of an antibiotic mixture previously determined to be active against the fecal and vaginal flora of the mother. All of the newborn obtained by means of this technique, including 13 piglets, 2 lambs, and 4 human infants, were found to be axenic throughout their stay in the isolators. Four piglets obtained by the same technique, but without adminstration of antibiotic mixture, were found not to be anenic. This technique, as compared with methods of surgical delivery of axenic young, embodies a number of advantages. It is harmless to the mother and to the newborn, it is relatively inexpensive, and it obviates the risk of prematurity involved in elective surgical delivery before term.

A blind qualitative and quantitative study of the aerobic and anaerobic faecal flora of two groups of newborn babies was carried out. One group of babies was breast-fed, while the second group was bottle-fed. In both cases the first established bacterial flora was essentially made of Escherichia coli and group D Streptococci (S. faecalis or S. bovis). On the sixth day of life a difference between he bacterial flora of the two groups was noticed, Bifidobacterium spp. was more frequently observed in the breast-fed babies. However the number of Bifidobacterium was lower than the dominant flora.

A standardized methodology was used to study the in vitro behavior of different strains of Neisseria gonorrhoeae when treated with povidone-iodine. The bactericidal activity of the antiseptic was appraised using the dilution-neutralization technique on four gonococcal strains, two of which produced a beta-lactamase. Bactericidal activity in aqueous medium was observed down to a dilution of 1/128. This result specifies the conditions under which this antiseptic is active against Neisseria gonorrhoeae.

We have tried in this study to reproduce in germfree mice a bacterial equilibrium observed previously in the gastrointestinal tract of child "L.N.", reared in a plastic isolator. Eight bacterial strains were studied. Seven had been fortuitously introduced: Acinetobacter lwoffi LN1, Staphylococcus epidermidis LN2, Clostridium perfringens LN3, Plectridium anatum LN4, Streptococcus bovis LN5, Enterobacter cloacae LN6, Corynebacterium sp LN7. The eighth strain was purposely administered to the child in order to take advantage of an antagonism between this strain and spontaneously acquired flora. This strain is Bacillus licheniformis FD1. The gnotoxenic mice proved to be a good experimental model because we found their bacterial equilibrium comparable to that observed in child L.N. In addition, we studied the decontamination procedures of the gastrointestinal tract of mice by observing both the antagonism of B. licheniformis with bacitracin, and the action of a suitable mixture of antibiotics. Conditions for the realization of this decontamination are discussed in this study.