Rapid identification of Mycobacterium species.

Identification of Mycobacterium species is currently a long and fastidious procedure. We have developed a rapid (5-h) standard method using the API-ZYM system and rapid nitratase, urease and catalase tests. Pigmentation and growth rate were noted (but were only necessary for complete identification of 6% of strains). The tests were assigned numerical values from which a profile number was derived. A total of 716 strains were studied: 434 belonging to the M. tuberculosis complex and 282 other mycobacteria including 21 from M. avium complex. All M. tuberculosis complex strains were differentiated from all other mycobacteria and M. bovis was clearly separated from M. tuberculosis. All M. avium complex strains were differentiated from other mycobacteria. Among mycobacteria other than tubercle bacilli, 97% of the strains studied were identified. The method has proven to be simple, rapid and standardizable. It is suggested that the use of a code list could permit identification of most mycobacteria.

Specific interaction between Listeria monocytogenes and glycosylated albumins.

We have previously shown that Listeria monocytogenes serovar 1/2b can bind strongly to bovine albumin (BA) glycosylated by glucosamine or fucosylamine with about 20 to 30 carbohydrate residues per albumin molecule. We now show that the binding is time-dependent, reversible, saturable and specific. The two glycosylated compounds inhibit each other competitively. Scatchard analysis showed that about 100 molecules of BA-glucosamide (heptameric configuration) and 14,300 molecules of BA-fucosylamide (monomeric configuration) bound per bacterial cell. The apparent dissociation constants for BA-glucosamide and BA-fucosylamide were found to be 3.9 x 10(-14) M and 3.5 x 10(-13) M, respectively.

Surface Listeria monocytogenes carbohydrate-binding components revealed by agglutination with neoglycoproteins.

Carbohydrate-binding components were shown to be present at the surface of Listeria monocytogenes by means of a panel of neoglycoproteins using direct agglutination. These lectin-like components bind on neoglycoproteins bearing D-glucosamine, L-fucosylamine, or para-amino-phenyl-alpha-D-mannopyrannoside residues. The interactions were inhibited by the carbohydrate moieties specific to the neoglycoproteins. The protein nature of the lectin-like components of L. monocytogenes was ascertained by the loss of carbohydrate-binding capacity following protease treatment.

Surveillance of Mycobacterium tuberculosis drug resistance in France, 1995-1997. AZAY Mycobacteria Study Group.

OBJECTIVE: To measure the rate of primary and secondary drug resistance of Mycobacterium tuberculosis on an ongoing basis. DESIGN: Data on all culture-positive tuberculosis were collected prospectively from 1995 through 1997 from a microbiological laboratory network of 19 university hospitals throughout France, and submitted quarterly to the National Reference Centre for Surveillance of Mycobacterial Diseases. RESULTS: A total of 2998 patients were included in the study. Among the 2333 (78%) previously untreated patients, 8.6% had isolates resistant to any drug, 4.8% to streptomycin (SM) alone, 1.2% to isoniazid (INH) alone, 1.8% to SM + INH, and 0.3% to INH + rifampicin (RMP) or multidrug resistance (MDR). Foreign birth was independently associated with a higher risk of primary resistance to any drug (odds ratio [OR] 1.5). Among the 268 (9%) previously treated patients, 20.9% had isolates resistant to any drug, 6.3% to SM alone, 3.4% to INH alone, 4.1% to SM + INH, and 3.7% to INH + RMP. Foreign birth (OR = 2.3), and human immunodeficiency virus positive status (OR = 4.4) were independently associated with a higher risk of secondary resistance to any drug. CONCLUSION: During the last 30 years there has been no increase in resistance to any drug among previously untreated patients. As expected, secondary resistance was highly associated with foreign birth. MDR-TB remains a rare event in France.

[Epidemic of listeriosis in Western France (1975--1976)]. / Epidémie de listériose dans l'Ouest de la France (1975--1976).

Cases of human listeriosis have recently increased in Western France (Maine-et-Loire and Mayenne); until 1974, there were only 15 documented cases yearly where as in 1975, there were 113 cases, and in 1976, 54 documented cases at the hospitals of Angers and Laval. Included amongst the clinical manifestations observed, were 126 cases of materno-fetal infections and 41 cases of either septicemia or meningo-encephalitis. The bacteriological study revealed a certain variability of biological characteristics examined, the overwhelming frequence of bacteria of the strain 4 b and the predictable sensitivity of this strain to antibiotics commonly employed in cases of listeriosis. The human cases were notably more frequent between January and June but their geographical distribution was not related to cases of either animal disease nor listeria that was isolated from corn silage. In this report, we propose an etio-pathological explanation for this epidemic.

[Cutaneous ulcer from Mycobacterium ulcerans. Apropos of 1 case in French Guiana]. / L'ulcère cutané à Mycobacterium ulcerans. A propos d'une observation en provenance de Guyane française.

Skin ulcer due to Mycobacterium ulcerans is presented. The patient come from East of French Guyana. Growth of this mycobacteria is obtained with diphasic Lowenstein medium at 30 degrees C. Diagnostic of M. ulcerans results from mycolic acids study.

[Study of antigen HBs and antivirus antibodies of hepatitis C during hepatopathies in Mali]. / Etude de l'antigène HBs et des anticorps anti-virus de l'hépatite C au cours des hépatopathies chroniques dans des services hospitaliers a Bamako, Mali.

A prospective study carried out in Bamako, Mali between July 1998 and January 1999 has assessed the seroprevalence of hepatitis C virus (HCV) in 91 carrier patients of chronic hepatopathy at a cirrhrosis stage (53) or of hepato-cellular carcinoma (38) and to compare with in 92 blood donors as a control population. Only seroprevalence confirmed by a complementary test has been taken into account (RIBA). HCV seroprevalence reached 25% including all hepatopathies, 24% in cirrhrosis and 26% in hepato-cellular carcinomae (HCC) versus 4% in blood donors. Antigen HBs of hepatitis B virus has been found in 55% of patients, versus 25% of the control cases (p = 0.0006). On the whole, the two markers have been notified a little more often in HCC than in cirrhosis and the combination of the two markers has been more frequent during cirrhosis as well. The role of HCV played in cirrhosis and HCC onset in Mali appears to be important.

[Bacteriological diagnosis of tuberculosis: current hieratic classification of methods]. / Diagnostic bactériologique de la tuberculose: hiérarchisation actuelle des méthodes.

To assure the diagnosis of tuberculosis, one needs the observation, the isolation and the identification of the causative agent Mycobacterium tuberculosis. In this approach, the microscopic exam occurs as a fast but neither sensitive or specific test. The isolation on solid media is slow and needs more than three weeks before becoming positive. Nevertheless, it is a sensitive and specific one. The identification of the isolated strain and the study of sensitivity to antibiotic agents require an equal delay. Then, 2 months are necessary to achieve the analysis. The AIDS epidemic with the increase of opportunistic mycobacterial diseases, and the unexpected arrival of resistant Mycobacteria is creating as a difficult therapeutic problem. The cultivation in liquid media with the radiometric method (Bactec) shortens the time of culture by half. The genomic amplification assay has been hopeful because it allowed results in 2 days. However, some technical difficulties happen when the test is conducted and it is less sensitive than the isolation process. The hierarchical classification of the laboratory useful process to establish the diagnosis of tuberculosis disease remains the microscopic observation of the bacilli and their isolation. Today, the use of PCR alone does not assure the diagnosis of tuberculosis, however it may be used as a additional diagnostic test.

[Severe pneumonia with a pneumococcal aspect during an ornithosis outbreak]. / Pneumopathies sévères d'allure pneumonique, au cours d'une épidémie d'ornithose.

OBJECTIVES: To describe the clinical, radiological and biological features of Chlamydia psittaci pneumonia. METHODS: A pneumonia outbreak occurred in a healthy middle-aged population working in a poultry slaughterhouse. Systematic serology (2 samples at 5 weeks intervals) provided the diagnosis of Chlamydia psittaci pneumonia in 6 patients. Patient files were analyzed retrospectively. RESULTS: The clinical presentations in this series of pneumonia were particularly homogeneous with a pneumococcal profile in all 6 cases: sudden onset, temperature above 39 degrees C, lobar alveolar involvement, hypoxemia, hyperleukocytosis and liver dysfunction. One case of hallucinatory delirium was observed. The patients were given spiramycin (9 million units per day for 3 weeks) and all recovered rapidly with no complications. CONCLUSION: The unusual virulence of the Chlamydia psittaci and very important inoculum were probably involved in this outbreak because of the severity of the pulmonary features and the short exposure of some patients to the bacteria. These cases suggest that the prevention of ornithosis in poultry slaughterhouses should be reinforced.

[Cervicofacial infections due to atypical mycobacteria and tuberculous lymph nodes]. / Infections cervico-faciales à mycobactéries atypiques et tuberculose ganglionnaire.

Cold abscesses in the cervical region, without associated general and functional signs, may be tuberculous lymph nodes or infected foci due to atypical mycobacteria. In the former, long-term antibiotic therapy, possibly followed by surgery conducted under conditions applicable to curettage for these lesions, is formally indicated. Treatment is of only moderate value in the second case, and spontaneous healing without other than esthetic sequelae usually occurs. These two lesions are quite different, therefore, but this was not recognized, and confusion still exists in many cases, for two reasons. Firstly, acid-alcohol-resistant bacilli are found in the pus in both cases, and a distinction cannot be made between them without culture. Secondly, pathological examination of specimens removed demonstrates identical appearances. The only way to distinguish between the two lesions, and to avoid excessively long or aggressive treatment, is by means of bacterial culture.

[The role of cytomegalovirus in infantile deafness and deafness of unknown origin]. / Le Rôle du cytomégalovirus dans la surdité du nourrisson et sa place dans les surdités d'étiologie méconnue.

It is widely accepted that the cause of congenital deafness is genetic in one third of cases roughly, is due to acquired affections during pregnancy or delivery in another third and remains unknown in the last third. It is possible that the cytomegalovirus (CMV) plays an important role in the latter group. The CMV is thought to be involved in 10 to 30% of cases of auditory sequelae from fetal infection, either severe neonatal CMV-induced disease, which is rare, or the frequent subclinical infections affecting an average of 1% of newborn infants. The only certain way to determine the importance of the role of CMV in deafness of unknown etiology is large-scale neonatal biologic screening followed by long-term audiologic surveillance: currently available documented data suggest that this role is very important.

Sentinel-site surveillance of Mycobacterium avium complex pulmonary disease.

The incidence of Mycobacterium avium complex (MAC) pulmonary disease in HIV-negative patients was studied prospectively from January 1, 2000 to December 31, 2002 through 32 sentinel sites distributed all over France. Among the 275 patients who yielded MAC isolates from respiratory clinical specimens, 101 (36.7%) met the bacteriological, radiographical and clinical criteria established by the American Thoracic Society for nontuberculous mycobacterial respiratory disease. Of these 101 patients, 81 had underlying lung disease, mainly previous tuberculosis, bronchectasis or chronic obstructive pulmonary disease. Among the 20 patients with no underlying lung disease, 12 had a predisposing factor such as leukaemia or immunosuppressive treatment and eight had no predisposing factor. All patients with MAC respiratory disease had clinical symptoms, commonly cough and fatigue, and 52 (51.5%) were sputum smear positive for acid-fast bacillus. The ratio of patients with Mycobacterium avium complex pulmonary disease to patients with pulmonary tuberculosis in France was estimated to be 3% and the incidence of Mycobacterium avium complex pulmonary disease in France was 0.2 per 100,000 inhabitants.

[Not Available].

Summary A cross-sectional study led in Bamako analyzed the seroprevalence of hepatitis C virus (HCV) and its genotypes among 91 patients carrying chronic liver diseases at the stage of cirrhosis (53) or hepato cellular carcinoma (38) and, on comparative basis in 92 blood donors as control population. False serologic reactions were found with ELISA (3/91 either 3,3% of the liver diseases and 1/92 or 1,1% of the control). Positive tests by ELISA confirmed by a RIBA test were finally considered. Concerning all the liver diseases, the seroprevalence of HCV was 15,4% including 15,1% in cirrhosis, 21% in hepatocellular carcinoma patients versus 2,2% in blood donors. The HBs antigen was associated in 5,6% of the cases In the hepatite C population, genotype 2a/2c was definitely prevalent, about 85,7%. Thus the role of the HCV in genesis of cirrhosis and hepatocellular carcinoma in Mali, appears significant.

Efficacy of the combination rifampin-streptomycin in preventing growth of Mycobacterium ulcerans in early lesions of Buruli ulcer in humans.

Mycobacterium ulcerans disease is common in some humid tropical areas, particularly in parts of West Africa, and current management is by surgical excision of skin lesions ranging from early nodules to extensive ulcers (Buruli ulcer). Antibiotic therapy would be more accessible to patients in areas of Buruli ulcer endemicity. We report a study of the efficacy of antibiotics in converting early lesions (nodules and plaques) from culture positive to culture negative. Lesions were excised either immediately or after treatment with rifampin orally at 10 mg/kg of body weight and streptomycin intramuscularly at 15 mg/kg of body weight daily for 2, 4, 8, or 12 weeks and examined by quantitative bacterial culture, PCR, and histopathology for M. ulcerans. Lesions were measured during treatment. Five lesions excised without antibiotic treatment and five lesions treated with antibiotics for 2 weeks were culture positive, whereas three lesions treated for 4 weeks, five treated for 8 weeks, and three treated for 12 weeks were culture negative. No lesions became enlarged during antibiotic treatment, and most became smaller. Treatment with rifampin and streptomycin for 4 weeks or more inhibited growth of M. ulcerans in human tissue, and it provides a basis for proceeding to a trial of antibiotic therapy as an alternative to surgery for early M. ulcerans disease.

Persistence of Listeria monocytogenes in three sorts of soil.

Life time of Listeria monocytogenes (strain PS 10401, serovar 4b) was studied in three sorts of soil: a chalky soil, poor in organic matters (pH 8.3) a peaty soil rich in organic matters (pH 5.5) a mixture of a chalky and peaty soil (pH 7.9). About 10(5) colony forming units (c.f.u.) per g of dry material were inoculated to each sort of soil which was incubated at 4 degrees C or at 20 degrees C. Periodically samples were taken and a numeration of germs was made on Tryptose Soy Agar. When direct numeration was negative, an enrichment was made at low temperature. In peaty soil, L. monocytogenes disappeared after incubation of 162 days at 4 degrees C and 156 days at 20 degrees C. In chalky soil, L. monocytogenes disappeared after 394 days of incubation at 20 degrees C, whereas at 4 degrees C, 10(4) c.f.u./g of dry matter were found 1500 days later. In mixture of chalky and peaty soil, 10(4) c.f.u. were found 1500 days after inoculation at 4 degrees C whereas 10(3) c.f.u. were found at 20 degrees C. Acid soils do not allow the persistence of L. monocytogenes for more than 160 days. Low temperature are significantly favourable to the persistence of L. monocytogenes.

Preservation of the virulence of Listeria monocytogenes in different sorts of soil.

Listeria monocytogenes (strain PS 10401, serovar 4b) whose virulence was well known, has been placed for 600 days in a sterile chalky soil (pH 8.3) and in a mixture of chalky and peaty soil (pH 7.9) and incubated at 4 degrees C and 20 degrees C. Groups of six OF1 mice were challenged by subcutaneous injection in the rear left hind foodpad with each sort of germs. Three days after inoculation, mice were killed by cervical disruption and numeration of L. monocytogenes was made in the spleen. Chemical composition of soil does not change virulence of germs. But low temperatures significantly increase virulence.

A phage typing system for Listeria monocytogenes and its use in epidemiological studies.

A phage-typing system for Listeria monocytogenes is necessary to eludicate its transmission and spread. We describe the general characteristics of a phage-typing system for L. monocytogenes and its use in selected epidemiological studies. Previously 78.4% of 823 strains of Listeria isolated in France between 1958 and 1978 were typable with a set of twenty phages isolaged from lysogenic strains. We now use twenty-seven phages; six derived from serovars 1/2a and 1/2b, eight from 4b, and thirteen from 4ab, 3c, 5, 6a and 6b. We used this system to examine 186 strains isolated during and after an outbreak in the province of Anjou which occurred during 1975 and 1976. One hundred and eight of these strains were of one phage type (octal code 000 420 000). Using this system we confirmed two cases of cross-infection in hospital neonates in England. Our results show that the reproducibility of the technique and the discrimination of the system are satisfactory for routine use. However, further testing and development of this system are underway.