[Methods of in vitro study and classification of antiseptics and disinfectants].
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Biomedical subjects
Publications and source records attributed to J Fleurette.
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A comparison has been done between various characters of 170 non epidemic strains cultivated from hospitalized patients and 29 strains originated from healthy carriers without any hospital contact and antibiotic treatment since 6 months. Following characters have been tested: M.I.C. of 13 antibiotics, heterogenous methicillin resistance, enzymatic antibiotic resistance, macrolide inducible or constitutive type resistance, mercuric chloride susceptibility; phage types; enzymes and toxins production. Antigenic structure has been studied, in particular production of protein A, A beta teichoc acid and type antigens. Significant differences have been observed between hospital and healthy carriers strains; these later strains are antibiotic susceptible, produce few toxins and enzymes, are not frequently phage typable. Their antigenic structure shows also significant differences: A beta-teichoic acid is less frequently detected; type antigens are less frequent and different. Role of these surface antigens in virulence is discussed.
The results of a continuous series of 518 adults undergoing cardiopulmonary bypass surgery, operated on by the same surgeon, comprising valvular replacement and aorto-coronary bypass surgery with a peroperative protocol of asepsis and a short prophylactic course of Penicillin and Streptomycin, a preoperative and postoperative bacteriological study, were treated by computer. The postoperative temperature chart showed a progressively decreasing pyrexia in the first 8 days after cardiopulmonary bypass. The nature of the operation, the bypass time, the quantity of blood used during operation and the blood loss were statistically significant factors. Patients with a clinical infection had significantly higher temperatures from the first on. 8,9% of patients had a febrile reaction which continued after the 10th day after cardiopulmonary bypass. The cause was not always apparent but the appearance of their temperature graphs was distinguishable after the first week. Analysis of the systematic bacterial specimens showed bacterial contamination of nearly 10% of drains, 10% of blood cultures, over 15% of intravenous infusion catheters and 70% of urinary catheters. The significance of these results is discussed. The normal appearance of the temperature chart after cardiopulmonary bypass surgery in the absence of clinical complications and bacterial contamination has been established.
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225 control subjects, 12 hospitalized Staphylococcus free (34 sera) and 16 S. aureus infected patients (79 sera) were tested for anti-A beta teichoic acid antibodies, using counter-immunoelectrophoresis. Anti-alpha and gamma haemolysins antibodies were dosed in parallel in 164 sera. Less than 5 per cent of control sera (3,11 per cent) have an antibody tire higher than 1/8: this tire is therefore selected as threshold level of positivity. Ten of the 12 hospitalized patients without staphylococcal infections are below this level. 31,6 per cent of staphylococcus infected patients have higher serum titers; only 3,7 per cent have no antibodies. In the others, a significant rise in titres is observed. There is no correlation between anti-alpha and gamma staphylolysin and anti-teichoic acid antibodies titres. The method suggested, both easy and rapid, could be used in association with anti-staphylolysin dosage for serological diagnosis of staphylococcal infections.
Staphylococcus aureus methicillin resistants strains cultivated on subinhibiting methicillin concentrations are not modified as concern protein A and type antigen 18. Low protein A production of these resistant strains known to be heterogenous as concern their resistance to methicillin does not correspond to absence of protein A on the highly resistant fraction of the population.
878 strains of Streptococcus pneumoniae were tested during two periods : 667 strains from 1972 to 1977 and 211 strains during 1978. Strains were isolated from spinal fluids, blood, peritoneal fluids, urines, exudate from respiratory tract except sputum. In polymicrobial specimens, only the predominant species were serotyped. From 1972 to 1977, 53,3% of strains are typed and 6 serotypes and pools are prevalent among 68,5% of typed strains, the most frequent are : 6 - D - 18 - 19. During the second period (1978) 82,5% of strains are typed, 9 serotypes are prevalent among 68,5% of typed strains, the most frequent are : 6 - 19 - 23 - 3. During 1978, 27,6% of strains are isolated from child wards, others from adults wards. A comparison of the relative frequencies of the serotypes in the two wards shows that the following 10 serotypes are the most frequent: 6 - 19 - 23 - 3 - 18 - 1 - 8 - 9 - 14 - 4.
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The specific Staphylococcus aureus agglutinogen h1 has been purified and shown to be a protein with a molecular weight of about 95,000. Chemical analysis revealed all the common amino acids, except tyrosine and the sulphur-containing ones. The purified h1 antigen was strongly immunogenic in rabbits. The antiserum produced one precipitation line on double diffusion in agar against a suspension of bacteria. It also agglutinated bacteria of the h1-containing strains, as well as tanned sheep erythrocytes sensitized with h1, in high dilutions. Antibodies to other known staphylococcal antigens were not detected. The identity between h1 and Pillet's type 9 antigen was confirmed.
We have endeavored to elaborate a suitable method for easy and rapid identification in clinical microbiology laboratories of the different species of infection-inducing, coagulase-negative staphylococci. Ten type strains described by Kloos and Schleifer and 269 strains isolated from 95 patients were tested; the classical tests were used for determination of Staphylococcus species. Strains were identified by using the Kloos-Schleifer reference method and the micromethod simultaneously. After preliminary tests on 77 substrates, 19 were retained, 15 for determination of species and 4 to reveal biotypes. The substrates were placed in wells in a rigid strip of inert plastic. Inoculation of wells was carried out with rich microbial suspensions in a special medium; reading of substrate reactions was done after incubation for 48 h at 35 degree C. The intrasystem reproducibility was excellent, from 91 to 100% for the 19 substrates. It was in excellent agreement with the reference method, 100% for type strains and 97.9% for hospital-isolated strains. Because it is simple and easy to reproduce, the micromethod will be most useful in clinical and ecological microbiology laboratories.
Using passive haemagglutination test, 48 human strains of Staphylococcus aureus cultivated on trypticase soya have been shown to be non-producers, but after subcultures especially on gelified "Oxoid broth n 2", 46 strains (95%) produced protein A. This observation points out the troubles encountered during protein A detection on the field of routine work.
This study describes the reproducibility of antigen detection in staphylococci, and the discriminating capacity of the serotyping method. The reproducibility of the antigen detection in staphylococci has been studied by multiple determination of the antigenic formula of 700 wild strains. The discriminating capacity of the serotyping method has been estimated on 415 strains selected to exclude strains from the same origin. Good reproducibility (over 80%) is obtained for the detection of 12 antigens; improvement of the method for the others is under way. The method is highly discriminative; only 1.4% of strains are not typable. 90.8% of strains can be classified in one of the 6 groups identified by a major antigen; groups A, D and E are the most important under our ecological conditions.
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The bacterial flora of the skin of the fore-arm was studied in fifteen normal subjects by a modified Williamson and Kligman technique. The quantitative results were in agreement with those found in other laboratories which indicates the reproducibility of the method. The action of an antiseptic containing 0.5 p. cent of benzalkonium chloride was studied under three conditions or single application of the antiseptic for five consecutive days and for 24 hours after the application of an occlusive dressing. The bacterial flora of the skin showed marked quantitative variations from one subject to another and, in the same subject, from one time to another. Evaluation of the activity of an antiseptic should take into consideration these physiological variations and also various conditions of use in practice. Are discussed also the factors leading to standardisation of the methods and the criteria of evaluation of the activity of antiseptics on the skin.