[Multiple erythema annulare centrifugum associated with knee monoarthritis revealing Capnocytophagacanimorsus infection]. / Erythème annulaire centrifuge multiple et monoarthrite révélateurs d'une infection à Capnocytophaga canimorsus.

INTRODUCTION: Capnocytophagacanimorsus (C. canimorsus), a commensal Gram-negative bacillus found in the oral cavity of dogs and cats, is pathogenic for humans, with the most common clinical manifestations being septicemia, meningitis and endocarditis. Herein we report a case of CC bacteremia manifesting as multiple plaques of erythema annulare centrifugum associated with monoarthritis of the knee. PATIENTS AND METHODS: A 66-year-old man consulted for a skin rash and monoarthritis of the right knee with fever following an insect bite on his right hallux. Cutaneous examination revealed numerous erythematous annular plaques on the trunk and limbs with centrifugal extension. Analysis of synovial fluid from the right knee showed an inflammatory liquid with a sterile bacteriological culture and PCR was negative for Borrelia. C. canimorsus bacteria were isolated from blood cultures. 16S RNA PCR performed on the synovial fluid was positive for the same organism. The patient's history revealed that his hallux wound had been licked by his dog. DISCUSSION: C. canimorsus most frequently affects immunosuppressed subjects. Cutaneous signs are seen in half of all cases, most frequently presenting as cellulitis, pathological livedo or thrombotic purpura. We report herein a case of CC bacteremia in an immunocompetent patient manifesting as multiple plaques of erythema annulare centrifugum, an unusual sign, and monoarthritis of one knee.

Human babesiosis in Alsace.

OBJECTIVES: Human babesiosis is a rare parasitic anthropozoonosis transmitted to humans by tick bites. Fifty-six cases of human babesiosis have been recorded in Europe. Two cases of babesiosis were reported in Alsace, France, in 2009. We performed a retrospective observational descriptive study to assess the epidemiology of the disease in Alsace. METHODS: Patients were included if they had a positive serology result for Babesia and/or a positive blood smear and/or a positive PCR result. The tests were performed in the microbiology laboratories of the university hospitals of Strasbourg, the civil hospitals of Colmar, and the hospital of Mulhouse between January 1, 2005 and December 31, 2015. Included patients were divided into three groups: definite case group (positive PCR or positive blood smear or seroconversion), possible case group (positive serology results without seroconversion with a compatible clinical picture and without other confirmed diagnoses), and incompatible case group (positive serology results without seroconversion, without compatible clinical picture and/or with other confirmed diagnoses). The compatible clinical picture was defined by the presence of flu-like symptoms and fever (≥38°C). RESULTS: Fifty-one patients had at least one positive result. Three cases were excluded (missing files). There were six definite cases, 12 possible cases, and 30 incompatible cases. All patients in the definite case group were immunocompetent. No deaths occurred. CONCLUSIONS: Human babesiosis is probably underdiagnosed due to its non-specific symptoms, lack of awareness about the disease, and the difficulty in making a diagnosis.

Analysis of delays in the prescription of oseltamivir in hospitals and potential for improvement.

OBJECTIVES: Patients hospitalized for influenza should receive early treatment with a neuraminidase inhibitor. PATIENTS AND METHODS: We conducted a retrospective study of the prescription of oseltamivir during the 2016-2017 influenza epidemic among patients hospitalized for influenza confirmed by RT-PCR in the infectious disease department. RESULTS: Treatment with oseltamivir was initiated as recommended in 96% of hospitalized patients presenting with influenza. However, a delay in prescription was observed with only 18% of prescriptions made on the first day. The prescriptions were exclusively initiated in the infectious disease department. CONCLUSION: To improve the early prescription of oseltamivir during the influenza season, two recommendations are essential: oseltamivir availability in the emergency department pharmacy, awareness of physicians of the need to prescribe to any patient hospitalized for a lower respiratory tract infection treatment with a neuraminidase inhibitor upon admission to the emergency department.

Cerebrospinal fluid monocytes in bacterial meningitis, viral meningitis, and neuroborreliosis.

OBJECTIVE: Cerebrospinal fluid (CSF) leukocytes analysis is commonly used to diagnose meningitis and to differentiate bacterial from viral meningitis. Interpreting CSF monocytes can be difficult for physicians, especially in France where lymphocytes and monocytes results are sometimes pooled. PATIENTS AND METHODS: We assessed SF monocytes in patients presenting with microbiologically confirmed meningitis (CSF leukocyte count>10/mm3 for adults or >30/mm3 for children<2 months), i.e. bacterial meningitis (BM), viral meningitis (VM), and neuroborreliosis (NB). RESULTS: Two-hundred patients (82 BM, 86 VM, and 32 NB) were included. The proportions of monocytes were higher in VM (median 8%; range 0-57%) than in BM (median 5%; range 0-60%, P=0.03) or NB (median 5%; range 0-53%, P=0.46), with a high value overlap between conditions. CONCLUSION: CSF monocytes should not be used to discriminate BM from VM and NB because of value overlaps.

Microbiology of French military casualties repatriated from overseas for an open traumatic injury.

BACKGROUND: This study aimed to describe the microbiological epidemiology of repatriated French soldiers with an open traumatic injury, and to measure the proportion of multidrug-resistant bacteria (MDRB). METHODS: Retrospective study including all French soldiers repatriated in 2011 and 2012 in Parisian military hospitals for open traumatic injury. Results of clinical samples and MDRB screening were collected. The antibiotic susceptibility was assessed using the agar disk diffusion method. Characterization of resistance mechanisms was performed using PCR. Genotyping of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E) isolates was performed using rep-PCR. RESULTS: A total of 139 patients were included; 70% of them were repatriated from Afghanistan. At admission, 24/88 were positive for MDRB (28%), mainly ESBL-E but no carbapenemase-producing Enterobacteriaceae and vancomycin-resistant Enterococcus faecium were identified. Forty-five patients had lesion sample collection, and 28/45 had a positive culture. The most frequently isolated pathogens were Enterobacter cloacae, Pseudomonas aeruginosa, and Escherichia coli. For eight patients, a MDRB was isolated from the wound, mainly ESBL-E (7/8) but also one methicillin-resistant Staphylococcus aureus and one imipenem-resistant Acinetobacter baumannii. Among ESBL-E, the PCR evidenced the high prevalence of CTX-M15 enzymes. Rep-PCR performed on the 23 ESBL-producing E. coli isolates highlighted numerous profiles. CONCLUSIONS: Controlling the spread of ESBL-E is currently challenging for French Armed Forces. Despite any evidence of an epidemic clone, a high-level compliance with hygiene precautions is required throughout the chain of care to avoid cross contamination.

Contribution of high-performance liquid chromatography to the identification of some Corynebacterium species by comparison of their corynomycolic acid patterns.

Reverse-phase high-performance liquid chromatography of corynomycolic acids provided a specific pattern for each Corynebacterium species studied. These data suggest that a fast and reproducible procedure is now available for bacteriological identification at the genus and at the species level of corynomycolic-acid-containing bacteria. Mass spectrometry analysis of post-column collected fractions provided the order of elution of some corynomycolic acids and isomers and showed the high specificity of the chromatographic assay which could be used for the routine bacteriological identification of some species belonging to the genus Corynebacterium.

Evaluation of the applicability of amplified rDNA-restriction analysis (ARDRA) to identification of species of the genus Corynebacterium.

The 16S rRNA genes (rDNA) of 50 strains belonging to 26 different coryneform bacterial species and genomospecies and of the type strain of Rhodococcus equi were enzymatically amplified. Amplified rDNA restriction analysis (ARDRA) with the enzymes AluI, CfoI and RsaI was carried out. The combination of the ARDRA patterns obtained after restriction with these three different enzymes enabled the differentiation between the following species: Corynebacterium accolens (number of strains = 2), C. afermentans subsp. afermentans (2), C. afermentans subsp. lipophilum (2), C. amycolatum (3), CDC coryneform group ANF-1-like (1), CDC coryneform group ANF-3-like (1), C. cystitidis (1), C. diphtheriae (4), C. jeikeium (3), C. macginleyi (2), C. minutissimum (1), C. pilosum (1), C. pseudotuberculosis (2), C. renale (2), C. striatum (2), C. urealyticum (3), C. xerosis (1), CDC coryneform groups B-1 (2), B-3 (2), F-1, genomospecies 1 and 2 (6), G, genomospecies 1 (1) and G, genomospecies 2 (2). The following strains or species could not be differentiated from each other: C. pseudodiphtheriticum (2) from C. propinquum (former CDC coryneform group ANF-3) (2), CDC coryneform group F-1, genomospecies 1 (4) from genomospecies 2 (2) and C. jeikeium genomospecies A (1) from genomospecies C (2). ARDRA may represent a possible alternative for identification of coryneforms, since this technique enabled the identification of most coryneforms tested and since DNA extraction (i.e. cell lysis by boiling), amplification, restriction and electrophoresis can be carried out within 8 hours. This might allow quick identification of C. diphtheriae and other possible pathogens of the genus Corynebacterium.

Corynebacterium group D2 ("Corynebacterium urealyticum") constitutes a new genomic species.

Twenty-one Corynebacterium group D2 ("C. urealyticum") strains were found to constitute a tight DNA hybridization group distinct from named Corynebacterium species. The strains of Corynebacterium group D2 had cell wall component type IV, short chain mycolic acids and G+C content of DNA of 65-66 mol %. Corynebacterium group D2 constitutes a genomic species which can be identified by phenotypic tests.

Taxonomy of Corynebacterium diphtheriae and related taxa, with recognition of Corynebacterium ulcerans sp. nov. nom. rev.

Levels of genomic DNA relatedness were determined using a S1 nuclease procedure for reference bacteria identified as biotypes of Corynebacterium diphtheriae, biovars of Corynebacterium pseudotuberculosis, and 'Corynebacterium ulcerans'. These results showed that the three species are separate taxa at the genomospecies level whereas biotypes and biovars are closely related genomically within each species. Phylogenetic analyses of small-subunit rDNA sequences revealed that 'Corynebacterium ulcerans' forms a tight cluster with Corynebacterium pseudotuberculosis within the robust branch that groups all Corynebacterium sequenced to date. Therefore, we propose that the species incertae sedis 'C. ulcerans' should be conclusively recognized as a distinct species within the genus Corynebacterium with strain CCUG 2708 = NCTC 7910 as type strain. This species is characterized by urease production and fermentation of glycogen.

Multiresistant corynebacteria in bacteriuria: a comparative study of the role of Corynebacterium group D2 and Corynebacterium jeikeium.

During an 11-month prospective study, urine cultures were performed on 5685 samples obtained in three hospital units. The use of a selective medium improved the recovery of antibiotic-multiresistant corynebacteria (AMC): 703 isolates (12.4%) compared with 88 isolates (1.6%) on sheep blood agar. Corynebacterium group D2 (CGD2) was isolated in 80.5% of urines yielding greater than or equal to 10(5) AMC ml-1 whereas Corynebacterium jeikeium represented 80.2% of isolates with less than 10(5) AMC ml-1. Among 16 patients with greater than or equal to 10(5) ml-1 C. jeikeium none had signs of urinary tract infection. In contrast, among 56 patients with greater than or equal to 10(5) CGD2, 40 (71%) had abnormal urinary sediment (mainly apatite or struvite crystals) and 29 (52%) had clinical signs of urinary tract infections sometimes complicated by lithiasis (seven cases) and alkaline-encrusted cystitis (two cases).

[Rhodococcus equi infection causing pulmonary malacoplakia in a patient with acquired immune deficiency syndrome]. / Infection à Rhodococcus equi responsable d'une malacoplasie pulmonaire chez un patient porteur d'un syndrome d'immunodéficit acquis.

Rhodococcus equi is a pathogen for some animal species. It can cause opportunistic pulmonary infections in immunocompromised people. The authors describe such an infection that causes malacoplakia in a patient with acquired immunodeficiency syndrome. The likeness between lesions due to Rhodococcus equi and those due to other opportunistic germs as Mycobacterium avium-intracellulare is emphasized.

Contribution of systematic RT-PCR screening for influenza during the epidemic season.

OBJECTIVE: We assessed the systematic RT-PCR screening of patients admitted to an infectious diseases department (IDD), during the 2012-2013 influenza outbreak. METHODOLOGY: Patients admitted with cough and fever underwent a nasopharyngeal smear for RT-PCR screening. RESULTS: Ninety-eight patients were admitted in the IDD, from January 1st to February 22nd, 46 were screened; 11 male and 6 female patients (17.3%, mean age of 68 years) were positive. The diagnoses made in the emergency department, before RT-PCR screening, were most frequently lung infection and sepsis, but influenza in only 4 cases. The diagnosis of influenza led to stopping antibiotics (n=4), initiating curative (n=4) and preventive (n=4) treatments with oseltamivir, and isolating patients to prevent a hospital outbreak. CONCLUSION: Systematic RT-PCR screening allows a rapid therapeutic management and the prevention of hospital epidemic through appropriate isolation measures.

[Evolution of antibiotic resistance of Streptococcus pneumoniae: results of Alsace observatory in 2005]. / Evolution de la sensibilité de Streptococcus pneumoniae aux antibiotiques: résultats de l'observatoire du pneumocoque Alsace (année 2005).

OBJECTIVES: Between 1st January 2005 and 31st December 2005, 232 strains of Streptococcus pneumoniae were collected in the Alsace county from participating laboratories (one from university hospital, 7 from general hospitals and 12 private laboratories) to assess their susceptibility to penicillin and evaluated serogroups of strains. METHOD: The coordinating centre performed MICs by the reference agar dilution test, interpreted according to CA-SFM breakpoints. Others antibiotics (erythromycin, cotrimoxazole, tetracycline...) were tested by agar diffusion, ATB-PNEUMO gallery or VITEK gallery (BioMérieux, France) by each participating laboratory. Data were processed, using 4th dimension software. RESULTS: Strains were collected from 151 blood samples, 38 ear pus, 11 cerebrospinal fluids, 8 pleural liquids and 24 representative pulmonary samples. The prevalence of pneumococci with decreased susceptibility to penicillin G (PDSP) is 35.1% (pulmonary samples excluded). The rate of PNSP decreases for all types of samples compared with other years of surveillance 2003 (44.0%). The rate of blood samples decreases for first time between the creation of Pneumococcal Observatory. The high-level resistance tend to decrease and began low. The PDSP are rather resistant to erythromycin, cotrimoxazole and fosfomycin. Among the PDSP, the most prevalent serotypes were 14, 19, 6 and 9. CONCLUSION: Among pneumococcal strains, the rate of PDSP tend however to decrease in 2005 compared with 2003. The rate stays inferior to the observed rates in other French counties where the same decreasing is described.

High-performance liquid chromatography of corynomycolic acids as a tool in identification of Corynebacterium species and related organisms.

A high-performance liquid chromatography (HPLC) study of 307 strains of Corynebacterium species and related taxa revealed that strains classified as "Corynebacterium aquaticum"; "Corynebacterium asperum"; and Centers for Disease Control (CDC) groups 1, 2, A-3, A-4, A-5, B-1, B-3, E, F-2, and I-2 as well as some unidentified coryneforms do not contain any corynomycolic acids; therefore, they should not be included in the genus Corynebacterium. Such an HPLC method of identification permitted the correct assignment to the genus Rhodococcus of two unpigmented strains of coryneform bacteria whose mycolic acid profiles were comparable to those of Rhodococcus equi. Bacteria belonging to CDC groups ANF-1, ANF-3, F-1, G-1, G-2, and I-1, as well as some other Corynebacterium sp. strains, yielded corynomycolic acid HPLC patterns related to those of Corynebacterium species. Either similarities or differences were observed in the corynomycolic acid profiles of Corynebacterium species tested after culture on sheep blood agar and/or sheep blood agar supplemented with Tween 80, which demonstrated that identification at the species or group level is possible. However, Corynebacterium striatum and CDC group I-1 bacteria as well as CDC group G-1 and group G-2 bacteria had indistinguishable HPLC patterns. Conversely, some variations were observed within some species as Corynebacterium xerosis, C. striatum, and Corynebacterium minutissimum. The evaluation procedure of this HPLC method by mass spectrometry analysis of isolated eluted peaks revealed that analytical reverse-phase HPLC alone does not provide any structural information, since isomers with identical polarities coeluted as a single peak. Nevertheless, HPLC is a rapid and reliable method for identification of corynomycolic acid-containing bacteria in the clinical microbiological laboratory.

Diagnostic value of the indirect immunofluorescence assay in cat scratch disease with Bartonella henselae and Afipia felis antigens.

Serum samples from 35 cat scratch disease (CSD) patients, 180 control patients (123 without lymph node enlargement and 57 with lymph node enlargement not evoking CSD), and 102 nonpatient subjects (35 with cat contact and 67 without cat contact) were tested by semiquantitative indirect immunofluorescence assay for the presence of antibodies directed to Afipia felis (ATCC 53690T) or Bartonella henselae (ATCC 49882T). The CSD group had statistically higher antibody titers against B. henselae than the control groups (P < 10(-5)), whereas no difference in A. felis antibody titers was evidenced among all groups tested. Among the 317 serum samples studied, the three with high A. felis antibody titers ( > or = 64) also had high antibody titers against other alpha-2 proteobacteria. The value of the indirect immunofluorescence assay with B. henselae antigen for the diagnosis of CSD was as follows: for a cutoff of 32, sensitivity was 0.80, specificity was 0.85, and the likelihood ratio was 5.1; for a cutoff of 64, the likelihood ratio was 12.1. In summary, in France, CSD is associated with high antibody titers against B. henselae, as previously described in the United States. However, the causes for B. henselae seronegativity in CSD patients and those for high antibody titers outside the typical nosological frame of CSD still have to be identified.

Mass spectrometry as a tool for identifying group D2 corynebacteria by their fatty acid profiles.

Corynebacterium group D2 (CGD2) are lipophilic antibiotic-multiresistant bacteria involved in some infections of immunocompromised patients. The fatty acid composition and structure of different strains was established by several mass spectrometric methods, particularly negative ion tandem mass spectrometry coupled with capillary gas chromatography. Non-hydroxylated fatty acid profiles of three strains of CGD2 (ATCC 43042, ATCC 43043, ATCC 43044) were almost identical and revealed the presence of several straight chain unsaturated fatty acids from the omega-9 series, with even carbon numbers ranging from 14 to 24. Branched saturated fatty acids were mainly anteiso-heptadecanoic acid and tuberculostearic acid. Surprisingly, a relatively large quantity of 10-methylene octadecanoic acid was found. The non-hydroxylated fatty acid profile of one rare beta-lactam susceptible strain (SC1) was different; 10-methylene octadecanoic acid was lacking whereas tuberculostearic acid was much more abundant. In contrast, the four CGD2 strains displayed highly similar mycolic acid patterns. The major mycolic acid species corresponded to C32, C30 and C28 bis-unsaturated with a double bond on each branch at the omega-9 position. The comparison of the mycolic acid composition and structure with those of other medically important corynebacteria strains, revealed a characteristic pattern for CGD2 strains, and CGD2 strains were easily distinguished from Corynebacterium jeikeium (CIP 82.51).

Genomic diversity among Corynebacterium jeikeium strains and comparison with biochemical characteristics and antimicrobial susceptibilities.

Levels of DNA relatedness were determined by performing DNA-DNA hybridization experiments (S1 nuclease procedure) with 13 human isolates exhibiting various antimicrobial susceptibility patterns which had been identified as Corynebacterium jeikeium by classical tests and the API Coryne system and with reference strains of C. jeikeium and related taxa. Twelve of 13 isolates which formed three genomic groups showed between 22 and 75% relatedness with the type strain of C. jeikeium. One of these genomic groups included all the strains resistant to penicillin and gentamicin and is genomically related to the C. jeikeium type strain at the species level. In addition, the reference strain of "Corynebacterium genitalium" biotype II was found to belong to this genospecies and therefore can be considered as a synonym of C. jeikeium. In contrast, one isolate and the reference strains of "Corynebacterium pseudogenitalium" biotypes C-3 and C-4 which were assigned to C. jeikeium by the API Coryne system were less than 10% related to the C. jeikeium type strain. These nongenomically related strains can be differentiated from the jeikeium-related strains on the basis of positive acidification from fructose and growth under anaerobic conditions. Furthermore, these strains exhibited full susceptibility to penicillin whereas the strains related to the C. jeikeium type strain are resistant to or only moderately susceptible to penicillin. No genomic relationship was found between C. jeikeium-related strains and other lipophilic coryneforms, identified as Corynebacterium accolens or Corynebacterium group G or F. Our study demonstrates the necessity to perform the fructose fermentation test or respiratory-type test for the correct identification of lipophilic coryneforms as C. jeikeium. Although these strains show genomic diversity at the species level, in a practical aspect, biochemical properties as well as antimicrobial susceptibility may allow the classification of such isolates in this single taxon.

Coryneform bacteria isolated from middle ear fluid.

Nineteen strains of facultatively anaerobic gram-positive rods isolated in pure culture from middle ear fluids were identified. All effusions were collected by tympanocentesis in children with acute otitis media. Identification of microorganisms to the genus level was done by studying the cell wall composition. Sixteen strains contain meso-diaminopimelic acid and arabinogalactan polymer but lack mycolic acids; therefore, these strains do not belong to a previously described taxon. Because of similarities with Corynebacterium afermentans (Centers for Disease Control group ANF-1), we temporarily classified these mycolateless strains ANF-1 like. Isolation of these microorganisms in pure culture from middle ear fluids collected by tympanocentesis is a strong argument for their involvement in acute otitis media.