Search

Asymptomatic faecal carriage of Clostridioides difficile has been widely evaluated, but its prevalence across a wide range of clinical departments and related risk factors are not well described. The objectives of the PORTADIFF study were to evaluate the prevalence and identifying risk factors leading to asymptomatic carriage of both toxigenic and non-toxigenic C. difficile.The PORTADIFF study was a 1-day prevalence study carried out in 10 different French hospitals. Adult patients, who agreed to participate, were included in this study and provided a fresh stool sample. C. difficile strains isolated from carriage were characterized by polymerase chain reaction (PCR) detection of tcdA, tcdB, cdtA and cdtB, and PCR ribotyping.In total, 721 patients were included in this study. The median age was 73 years (range 18-101 years) and the male/female ratio was 1.06. C. difficile (either toxigenic or non-toxigenic strains) was isolated from 79 (11%) patients; 42 (5.8%) strains were toxigenic. The prevalence rates of asymptomatic carriage ranged from 5% on surgical wards to 19% on long-term care wards. The main risk factors associated with asymptomatic carriage were antibiotic treatment within the preceding 3 months (81.8% vs 53.7%; P0.01), hospitalization within the preceding 2 months (55.8% vs 33%; P0.01), cumulative duration of hospital stay before study inclusion (mean 50.1 vs 34.5 days; P0.047), and hospitalization on a ward with high global incidence of C. difficile infection.Eleven percent of hospitalized patients were asymptomatic carriers of toxigenic or non-toxigenic C. difficile, and may constitute a potential reservoir of C. difficile strains.

BACKGROUND Trichosporonosis is a rare invasive infection in humans mainly due to Trichosporon asahii, and especially recovered from patients having haematological malignancy. Since 2012, IGS1 region sequencing is used as a genotyping method to distinguish isolates, with high frequency of one haplotype worldwide and a geographic specificity for some haplotypes. OBJECTIVES We compared the IGS1 genotyping method and whole genome sequencing (WGS) to study the relationship between clinical isolates involved in two grouped cases in France. METHODS IGS1 sequencing and antifungal susceptibility testing were performed for 54 clinical isolates. Clinical data for 28 isolates included in surveillance programs were analysed. Whole genome was sequenced for 32 clinical isolates and the type strain. RESULTS All isolates were intrinsically resistant to flucytosine, while voriconazole had the most potent in vitro activity. The majority of the isolates was recovered from patients with haematological malignancies (42.86%), with a high proportion of children (

Resume But de l’etude Clostridium difficile est le principal agent des diarrhees bacteriennes postantibiotiques et nosocomiales. Depuis l’emergence du clone NAP1/027 hypervirulent et epidemique en Europe, il apparait necessaire d’isoler les souches de C. difficile pour realiser un suivi epidemiologique par typage moleculaire. Ce travail a eu pour objectif de comparer trois conditions de culture selective pour l’isolement de C. difficile. Methodes Cent trente selles de patients hospitalises a l’hopital Jean-Verdier ont ete ensemencees sur le milieu commercial CLO (BioMerieux) et sur un milieu prepare au laboratoire (CCTa : Columbia, cefoxitine 8 mg/l, cycloserine 250 mg/l, sang de cheval 5 %, taurocholate de sodium 0,1 %) avec et sans choc alcoolique (EtOH) prealable. C. difficile a ete isole dans 38 selles et les colonies ont ete denombrees sur chaque milieu. Resultats L’intensite de fluorescence des colonies de C. difficile est comparable sur CLO et CCTa-EtOH, leur aspect etant plus caracteristique sur CLO. Ce milieu apparait tres selectif contrairement au milieu CCTa sur lequel une flore associee gene la lecture de la fluorescence et necessite le reisolement des colonies suspectes. Sur CCTa ± EtOH on denombre en moyenne 30 fois plus de colonies de C. difficile que sur CLO, suggerant une forte proportion de spores dans les selles. Conclusions Le milieu CLO est performant pour l’isolement de C. difficile malgre sa selectivite. Neanmoins, il apparait interessant d’associer un milieu favorisant la germination des spores tel que le milieu CCTa ensemence apres choc alcoolique, permettant d’augmenter la sensibilite de detection en s’affranchissant des conditions de conservation et de transport.

Resume Clostridium difficile est le principal agent responsable de diarrhees postantibiotiques nosocomiales. Depuis 2006, des foyers epidemiques ont ete decrits en France impliquant la souche hautement virulente NAP1/027 qui presente un haut niveau de resistance a l’erythromycine et aux fluoroquinolones. L’objectif a ete de suivre l’evolution epidemiologique et de surveiller la resistance aux antibiotiques de souches de C. difficile isolees dans le CHU Jean-Verdier–Rene-Muret dans des services medecine, chirurgie, obstetrique (MCO) et geriatriques. Au total, 33 souches toxinogenes isolees, entre 2001 et 2007, de patients symptomatiques ont ete etudiees. Les toxines A et B ont ete recherchees par methode immuno-enzymatique (ICTAB, Meridian). La sensibilite des souches vis-a-vis du metronidazole, de la vancomycine, de l’erythromycine, de la moxifloxacine a ete determinee par diffusion en milieu gelose. Les concentrations minimales inhibitrices (CMI) au metronidazole, a la vancomycine et a la tigecycline ont ete determinees par le E-test (AES). Pour la toxine binaire, les genes cdtA et cdtB ont ete recherches par PCR. Trente-deux souches ont fait l’objet d’un typage moleculaire par PCR-ribotypage utilisant les amorces decrites par Bidet et al. Sur la periode etudiee, aucune souche n’etait resistante au metronidazole, a la vancomycine et a la tigecycline. En revanche, on note une diminution de la sensibilite a la moxifloxacine (100 % en 2001 versus 28,5 % en 2007) et a l’erythromycine (60 % en 2001 versus 14 % en 2007). La PCR-ribotypage a permis de mettre en evidence : (1) la persistance d’un clone epidemique sur une periode de huit mois en 2002 ; (2) un clone epidemique chez sept patients d’un meme service de MCO entre juin et octobre 2007 et (3) des souches non reliees epidemiologiquement. A ce jour, aucune souche NAP1/027 n’a ete caracterisee parmi les souches resistantes a l’erythromycine et la moxifloxacine.

The in vitro activities of caspofungin and micafungin against 1,038 yeast isolates have been determined. The caspofungin and micafungin MICs were lower for Candida albicans , Candida glabrata , and Candida tropicalis than for Candida parapsilosis , Candida guilliermondii , and Candida krusei . A clear correlation was seen between the MICs for the two drugs.

Four immunoenzymatic tests for detecting Clostridium difficile toxins A and B were studied: two rapid tests (Tox A/B QUIK CHEK-Techlab and NoviView Toxine-A-Hiss diagnostics) and two Elisa tests (C. difficile TOX A/B II -Techlab and Toxin A+B Elisa Test, Novitec-Hiss diagnostics). The results were compared to those obtained with ImmunoCard Tox A+B -ICTAB (Meridian), C. difficile Toxine A (Oxoid) for rapid test and Elisa Premier A+B Meridian for Elisa. A total of 41 stools and 16 isolates were studied with rapid tests. On stools, the sensitivity and specificity of QUIK CHEK test was 94.1% and 100% respectively compared to the test ICTAB. On the isolates, sensitivity and specificity was 100%. With the Noviview test, the sensitivity on stools and isolates was respectively 88.2 and 85.7% and the specificity was 100% compared to Oxoid. A total of 38 stools were studied with Elisa tests. With Techlab test compared to the test Premier, sensitivity and specificity was 100%. The Novitec test gave five false negative reactions with consequently a sensitivity of 70.6%.

Resume Objectif. – Evaluer le niveau de resistance aux antibiotiques du pneumocoque en Ile de France. Methode. – En 2001, 637 souches cliniques de pneumocoque ont ete etudiees a partir de 32 laboratoires de microbiologie. Resultats. – Cinquante et un pour cent des souches ont ete isolees chez les enfants de moins de 15 ans et 49 % chez les adultes. Chez l'enfant 76 % des souches provenaient de pus d'oreille, 20 % d'hemocultures ; chez l'adulte la plupart des souches (92 %) provenaient d'hemocultures. Le pourcentage de pneumocoque de sensibilite diminuee a la penicilline G est globalement de 61 %, plus eleve chez l'enfant (73 %) que chez l'adulte (50 %). Parmi les souches de moindre sensibilite, 21,8 % sont resistantes a la penicilline (CMI > 1 mg/l). La sensibilite diminuee a l'amoxicilline et au cefotaxime concerne respectivement 38 et 17 % des souches. Les souches resistantes a ces deux molecules (CMI > 2 mg/l) sont rares 2,6 et 0,4 % respectivement. Parmi les autres antibiotiques, les taux de resistance sont de 63 % pour l'erythromycine, 47 % pour le cotrimoxazole, 40 % pour les tetracyclines et 23 % pour le chloramphenicol. Les serogroupes les plus frequents sont les serogroupes 19 et 14, respectivement 23 et 18 %. Les serotypes contenus dans le vaccin heptavalent couvrent 90 % des souches isolees chez l'enfant de moins de deux ans. Conclusion. – Les taux de resistance a la penicilline sont plus particulierement eleves dans les pus d'oreille chez l'enfant (76 %).

We report two cases of pregnancy-associated Plasmodium falciparum malaria discovered fortuitously. These two women were born in Africa and their last visit in an endemic area was more than one year before. It is well known that pregnancy is one of major risks of late onset of P. falciparum malaria. In the two cases reported in this study, clinical signs of malaria were not specific and we will describe the interest to detect more systematically pregnant African women, first arrival immigrants.

Resume En 1994, nous avons ete confrontes, a l'hopital Jean Verdier, a une epidemie de Klebsiella pneumoniae a beta-lactamase a spectre etendu (11 cas en 1994 pour 3 cas en 1993). Entre janvier et avril, ont ete repertories 3 cas sporadiques dont un suspect d'etre a l'origine d'une diffusion epidemique. A partir de cette periode, la succession des cas d'infection ou de portage nous a amenes a pratiquer une enquete epidemiologique. Les souches de K. pneumoniae ont ete isolees de prelevements a visee diagnostique ou epidemiologique. Elles ont ete comparees par leur biotype, leur antibiotype (phenotype de resistance aux beta-lactamines et aux aminoglycosides) et leur genotype (AP-PCR). Parallelement une enquete epidemiologique a ete entreprise (origine du patient, lieu et duree d'hospitalisation). Dans un premier temps, l'enquete epidemiologique nous a permis de mettre en evidence les contacts possibles entre les differents patients, dans les differents services. Dans un second temps, l'etude des souches a eu pour but de mettre en evidence une souche epidemique eventuelle. Toutes les souches ont le meme biotype. L'etude des phenotypes de resistance aux antibiotiques a permis de definir six groupes. Les profils d'amplification obtenus par AP-PCR ont permis de definir trois groupes de souches respectivement de genotype A, B et C. L'existence de souches de genotypes differents exclut la dissemination d'une seule souche epidemique dans l'hopital. En revanche, deux souches semblent responsables d'une diffusion hospitaliere : les souches de genotype A et B. Ces souches ont toutes ete isolees apres au moins 72 heures d'hospitalisation d'ou une acquisition probable dans l'hopital. La souche de genotype C a ete isolee d'un seul patient dans les 72 premieres heures de l'hospitalisation d'ou une importation probable de cette souche sans diffusion ulterieure.

The intestinal colonization by Enterobacter cloacae strains with a derepressed cephalosporinase was studied in a paediatric ward between February 1990 and January 1991. Environmental sampling was performed simultaneously. Fifty-two isolates were recovered from 200 neonates (stool, blood) and 14 strains were isolated from the neonatal environment. An epidemiological study based on the typing of 36 Enterobacter cloacae isolates was carried out using antibiotyping, biotyping and ribotyping methods. The isolates selected were from 21 neonates (24 isolates), the neonatal ward environment (8 isolates) and from other wards (4 isolates). Thirty-two isolates had the same antibiotic resistance pattern, corresponding to a derepressed cephalosporinase and resistance to the following aminoglycosides: kanamycin, gentamicin, tobramycin and netilmicin. No predominant biotyping pattern could be established. Ribotyping done with two endonucleases (EcoRI and BamHI) showed 28 Enterobacter cloacae isolates to have a single pattern. Ribotyping was the most discriminating method used in this study, permitting identification of cross-contamination with Enterobacter cloacae in the paediatric ward.

Preventive treatment of mice with trehalose 6,6' dimycolate (TDM), an immunomodulator of bacterial origin, enhances their resistance to encephalomyocarditis (EMC) virus infection. The protective effect of TDM is totally abolished by the injection of silica particles in mice, demonstrating the role of macrophages in the antiviral action of TDM. In vitro, peritoneal macrophages from mice treated with TDM (TDM-PM) exhibit an intrinsic antiviral activity against EMC virus, while resident peritoneal macrophages (RES-PM) are permissive to this virus. Greater amounts of interferon are detected in supernatants of cultures of TDM-PM than of RES-PM. Neutralization of interferon (IFN) by addition in vitro of anti-IFN alpha/beta serum markedly reduces the antiviral activity of TDM-PM. These results indicate that interferon alpha/beta is involved in the intrinsic anti-EMC virus activity of peritoneal macrophages from mice treated with TDM.

Clostridium difficile is the most common agent of postantibiotic and nosocomial bacterial diarrhoea. Since the emergence of the highly virulent and epidemic strain NAP1/027 in Europe, it appears necessary to isolate C. difficile strains to realize an epidemiologic follow-up by molecular typing. The aim of this work was to compare three selective culture conditions for the isolation of C. difficile.One hundred and thirty stools collected from patients hospitalized at Jean Verdier were swabbed on the commercial medium CLO (BioMérieux) and on a medium prepared at the laboratory (CCTa: Columbia, cefoxitine 8 mg/l, cycloserine 250 mg/l, horse blood 5 %, sodium taurocholate 0.1 %) with and without preliminary alcoholic shock (EtOH). C. difficile was isolated from 38 stools and colonies were counted on each medium.The fluorescence intensity of C. difficile colonies is comparable on CLO and CCTa-EtOH media, however their aspect is more characteristic on CLO. This medium appears very selective contrary to the CCTa medium on which an associated flora obstructs the fluorescence reading and requires a new isolation of the suspect strains. On average 30 times more colonies of C. difficile are counted on CCTa+/-EtOH than on CLO, suggesting the presence of great proportions of spores in the stools.The medium CLO is successful for the isolation of C. difficile despite of its selectivity. Nevertheless, it appears interesting to associate a medium enhancing spore germination as the CCTa medium inoculated after alcoholic shock to increase the sensitivity of detection while being freed from conservation and transport conditions.

We report two cases of pregnancy-associated Plasmodium falciparum malaria discovered fortuitously. These two women were born in Africa and their last visit in an endemic area was more than one year before. It is well known that pregnancy is one of major risks of late onset of P. falciparum malaria. In the two cases reported in this study, clinical signs of malaria were not specific and we will describe the interest to detect more systematically pregnant African women, first arrival immigrants.

Clostridium difficile is the most common agent of nosocomial bacterial diarrhoea in adults. In 2006, C. difficile outbreaks were described in France with the highly virulent strain PCR-ribotype 027, which is also resistant to moxifloxacin and erythromycin. The aim of this study is to perform a phenotypic and molecular characterization of C. difficile strains isolated in Jean-Verdier-René-Muret hospitals. Thirty three C. difficile toxigenic strains isolated in symptomatic patients from 2001 to 2007 were studied. Toxins A and B detection was performed with an immunoenzymatic method (ICTAB, Meridian). The agar diffusion method was performed for determination of antibiotic susceptibility for metronidazole, vancomycin, erythromycin and moxifloxacin. The E-test was performed for determination of metronidazole, vancomycin and tigecycline MIC. Binary toxin genes cdtA and cdtB were detected by PCR. PCR-ribotyping was performed according to Bidet et al. From 2001 to 2007, all the isolates studied were susceptible to metronidazole, vancomycin and tigecyclin. We observed a significant decrease of susceptibility to moxifloxacin (100% in 2001 versus 28.5% in 2007) and to erythromycin (60% in 2001 versus 14% in 2007). Toxins A/B were detected in all the isolates. Fifteen per cent of the isolates studied produced the binary toxin not correlated with a specific PCR-ribotype. Ribotype 18 was the most prevalent PCR-ribotype detected since 2006. The isolates displaying this PCR-ribotype were resistant to erythromycin and moxifloxacin and were principally isolated in the same ward, suggesting cross infection. This study showed that: (1) over a six-year period, the susceptibility to metronidazole and vancomycin remained stable; (2) different clones of C. difficile circulated during these six years. Recently an epidemic strain resistant to erythromycin and moxifloxacin of ribotype 18 has emerged in the gastroenterology unit where fluoroquinolones are frequently used demonstrating the role of antibiotic selection pressure. The emergence of these isolates could explain the significant decrease of susceptibility to moxifloxacin and erythromycin observed in 2007. However, today, no isolate with a PCR-ribotype 027 was detected.

In vitro determination of Clostridium difficile susceptibility to antibiotics is not routinely performed. The aim of this study was to evaluate the performance of antibiotic susceptibility determination with the disk diffusion method for screening C. difficile isolates with decreased susceptibility to antibiotics.Thirty-six C. difficile isolates (toxigenic or not) isolated in 2005 and 2006 from three hospitals Assistance publique-Hôpitaux de Paris (Jean-Verdier, René-Muret, Beaujon) were studied by disk diffusion method with 14 antibiotics. Mueller-Hinton agar supplemented with sheep blood (Bio-Rad*) were swabbed with a C. difficile suspension at 1 McFarland. To check the results obtained with the disk diffusion method, Minimal Inhibitory Concentration (MIC) were performed respectively with E-test for glycopeptides and metronidazole and with the agar dilution reference method and E-test for new molecules with a potential activity against anaerobes: imipenem, ertapenem, linezolid and moxifloxacin.The decreased susceptibility (resistant and intermediate) observed was 40% for amoxicillin-clavulanate, 60% for piperacillin-tazobactam, 100% for ceftriaxone, 81% for imipenem, 61% for ertapenem, 2% for chloramphenicol, 34% for erythromycin, 90% for lincomycin, 2% for linezolid, 98% for levofloxacin, 17% for moxifloxacin and 0% for vancomycin, teicoplanin and metronidazole. The results obtained with the disk diffusion method were compared to MICs obtained with E-test and reference method.The disk diffusion method seems to be a good method to detect isolates suspected to have a decreased susceptibility and consequently to reduce MIC determinations.

Our objective is to determine in vitro efficiency of moxifloxacin (MXF) alone or in combination with cefotaxime (CTX) on Group B streptococcus (GBS).For 21 strains of GBS isolated from newborn invasive infections (6 meningitis and 15 bacteraemia), the bacterial growth in Mueller Hinton broth with MXF and/or CTX leaded to the determination of MIC and MBC, the determination of tolerance for CTX and the evaluation of the bacteriostatic action of these antibiotics combination by calculating the FIC index. Time-kill studies were conducted for MXF and CTX alone or in combination for the first four hours, with concentrations likely reached in CSF.Study of GBS growth with crossed concentrations of MXF and CTX showed no resistant strains, no tolerant strains, and no antagonism between MXF and CTX. Killing curves demonstrated that MXF is ten-fold more active than CTX in the first four hours.MXF is an interesting antibiotic for its good activity on the GBS, suggesting that MXF is a good candidate for further evaluation in GBS meningitis in animal model.

Four immunoenzymatic tests for detecting Clostridium difficile toxins A and B were studied: two rapid tests (Tox A/B QUIK CHEK-Techlab and NoviView Toxine-A-Hiss diagnostics) and two Elisa tests (C. difficile TOX A/B II -Techlab and Toxin A+B Elisa Test, Novitec-Hiss diagnostics). The results were compared to those obtained with ImmunoCard Tox A+B -ICTAB (Meridian), C. difficile Toxine A (Oxoid) for rapid test and Elisa Premier A+B Meridian for Elisa. A total of 41 stools and 16 isolates were studied with rapid tests. On stools, the sensitivity and specificity of QUIK CHEK test was 94.1% and 100% respectively compared to the test ICTAB. On the isolates, sensitivity and specificity was 100%. With the Noviview test, the sensitivity on stools and isolates was respectively 88.2 and 85.7% and the specificity was 100% compared to Oxoid. A total of 38 stools were studied with Elisa tests. With Techlab test compared to the test Premier, sensitivity and specificity was 100%. The Novitec test gave five false negative reactions with consequently a sensitivity of 70.6%.

Congenital malaria is uncommon in France. The purpose of this report is to describe a case involving a six-week-old infant who was hospitalized with fever, hepatosplenomegaly, anemia and thrombopenia. Thick and thin blood smears were positive for Plasmodium malariae. The infant responded favorably to chloroquine. Based on this experience, we performed a search of the literature to find case reports on congenital malaria in France and compare clinical and epidemiologic data with series reported in the United States and from endemic areas. The placenta appears to provide an effective barrier against Plasmodium since infection is much more common than disease. The delay for onset of clinical symptoms is longer in temperate zones than in endemic areas. The type of parasite could account for this difference since African congenital malaria are due to Plasmodium falciparum while most cases described in the United States are due to Plasmodium vivax. We also discuss the possible implications of coinfection by HIV in the mother.

We report the case of a pregnant woman (29 th week), living in a Paris suburb, about 20 miles from an international airport. She presented with septic shock and severe acute respiratory distress syndrome (ARDS). A few parasitized erythrocytes were discovered in a hemorrhagic bronchoalveolar lavage (BAL), specimen and many were found on examination of the placenta after a caesarean section had been performed. The patient's condition dramatically improved once given quinine therapy. This is an uncommon case on account of: (1) the unusual clinical course with no organ failure but ARDS, (2) the unusual way the diagnosis was made, (3) the very unusual way the patient became contaminated (airport malaria), (4) the pregnant condition of the patient.

The authors wanted to assess the level of Streptococcus pneumoniae antibiotic resistance in Ile de France.In 2001, 637 clinical strains of S. pneumoniae were prospectively collected from 32 microbiology laboratories.Fifty one percent of strains were isolated from children under 15 years of age and 49% from adults. In children, 76% of strains came from otitis media, 20% from blood culture, in adults most strains (92%) came from blood culture. The overall prevalence of non-susceptible penicillin pneumococci was 61% higher in children (73%) than in adults (50%). Among the non-susceptible penicillin pneumococci 21.8% were resistant (CMI1 mg/l). Strains with decreased susceptibility to amoxicillin and cefotaxime were 38% and 17% respectively. Resistant strains to these two drugs (CMI2 mg/l) were rare 2.6% and 0.4% respectively. Among other antimicrobial agents, rate of resistance was 63% to erythromycin, 47% to cotrimoxazole, 40% to tetracycline, and 23% to chloramphenicol. The most frequent serogroups were serogroups 19 and 14, respectively 23% and 18%. Serotypes included in heptavalent vaccine covered 90% of children strains under 2 years of age.The prevalence of resistance to penicillin was high in children particularly in otitis media pus (76%).

A rapid commercial agglutination test (Bactigen Strepto B) for detection of group B streptococci in gastric aspirates of neonates was evaluated. One hundred and sixty-one gastric samples were analyzed with conventional bacteriological techniques and with the commercial test after modification of the extraction technique. The sensitivity of the test relative to the culture technique was 90.4 %, the specificity 94.2 %, the positive predictive value 70.3 % and the negative predictive value 98.5 %. The commercial test could be performed in one hour and showed good sensitivity and specificity. If a test result was negative colonization could be excluded, obviating the need for empirical antibiotic therapy, whereas a positive result suggested colonization or neonatal infection with group B streptococci.

The treatment of intestinal Clostridium difficile infections rests on administration of either a glycopeptide or metronidazole. Given the current shifts in resistance patterns of anaerobes to antimicrobials, a study of the susceptibility of C. difficile to metronidazole was timely. The objective of this study was to evaluate the influence of the culture medium on the Minimal Inhibitory Concentration (MIC) of metronidazole as determined using the E-test. Thirty-one strains were grown on three different media supplemented with 5% horse blood, namely Columbia agar, Wilkens Chalgren agar, and Brucella agar. Results were compared to those obtained using the reference agar dilution method (ADM). As recommended by the French Society for Microbiology, susceptibility was defined as an MICor = 4 mg/L. When used on strains susceptible by the ADM, the E-test yielded lower values than the ADM with all three media. Furthermore, findings suggest that E-test results obtained with strains whose MIC is in the 4 to 8 mg/L range by the ADM should be interpreted with caution and, in some cases, tested using the ADM.

The production of proteolytic enzymes by 10 Clostridium difficile isolates of varying toxigenicity and clinical origin was studied to determine if all isolates secreted proteases. Different protease substrates were studied: gelatin, collagen, phenylazobenzyloxycarbonyl-leucyl-glycyl-L-prolyl-D-arginine (Pz-peptide), casein, azocasein, and azocoll. All isolates degraded gelatin, collagen, and azocoll. The supernatants of all isolates contained an enzyme capable of attacking gelatin incorporated in a polyacrylamide gel (zymograms) and forming two closely spaced lytic bands with an estimated molecular mass of 35-40 kDa. Polyclonal antibodies, produced against the C. difficile gelatinase, revealed in Western blots a 35-kDa protein in the culture supernatants of all C. difficile isolates. In the same manner, Clostridium perfringens collagenase polyclonal antibodies detected a 120-kDa protein in the culture supernatants of all isolates; this suggests that at least two proteases may exist in C. difficile. The protease activities of the 10 strains examined did not seem strikingly different quantitatively but were in general weak and their role in pathogenicity is suspect.

Septic osteomyelitis of the hip in a previously healthy child is described. A weakly toxigenic Corynebacterium diphtheriae strain was isolated from the bone aspirate. The results of the treatment were rapidly satisfactory, after surgical drainage and antibiotic therapy with pristinamycin.This case report shows that C. diphtheriae has not disappeared in the developed world and can be responsible of systemic infections.

International audience; Since echinocandins are recommended as first line therapy for invasive candidiasis, detection of resistance, mainly due to alteration in FKS protein, is of main interest. EUCAST AFST recommends testing both MIC of anidulafungin and micafungin, and breakpoints (BPs) have been proposed to detect echinocandin-resistant isolates. We analyzed MIC distribution for all three available echinocandins of 2,787 clinical yeast isolates corresponding to 5 common and 16 rare yeast species, using the standardized EUCAST method for anidulafungin and modified for caspofungin and micafungin (AM3-MIC). In our database, 64 isolates of common pathogenic species were resistant to anidulafungin, according to the EUCAST BP, and/or to caspofungin, using our previously published threshold (AM3-MIC ≥ 0.5 mg/L). Among these 64 isolates, 50 exhibited 21 different FKS mutations. We analyzed the capacity of caspofungin AM3-MIC and anidulafungin MIC determination in detecting isolates with FKS mutation. They were always identified using caspofungin AM3-MIC and the local threshold while some isolates were misclassified using anidulafungin MIC and EUCAST threshold. However, both methods misclassified four wild-type C. glabrata as resistant. Based on a large data set from a single center, the use of AM3-MIC testing for caspofungin looks promising in identifying non-wild-type C. albicans, C. tropicalis and P. kudiravzevii isolates, but additional multicenter comparison is mandatory to conclude on the possible superiority of AM3-MIC testing compared to the EUCAST method.

Invasive yeast infections represent a major global public health issue, and only few antifungal agents are available. Azoles are one of the classes of antifungals used for treatment of invasive candidiasis. The determination of antifungal susceptibility profiles using standardized methods is important to identify resistant isolates and to uncover the potential emergence of intrinsically resistant species. Here, we report data on 9,319 clinical isolates belonging to 40 pathogenic yeast species recovered in France over 17 years. The antifungal susceptibility profiles were all determined at the National Reference Center for Invasive Mycoses and Antifungals based on the EUCAST broth microdilution method. The centralized collection and analysis allowed us to describe the trends of azole susceptibility of isolates belonging to common species, confirming the high susceptibility for Candida albicans (n = 3,295), Candida tropicalis (n = 641), and Candida parapsilosis (n = 820) and decreased susceptibility for Candida glabrata (n = 1,274) and Pichia kudriavzevii (n = 343). These profiles also provide interesting data concerning azole susceptibility of Cryptococcus neoformans species complex, showing comparable MIC distributions for the three species but lower MIC(50)s and MIC(90)s for serotype D (n = 208) compared to serotype A (n = 949) and AD hybrids (n = 177). Finally, these data provide useful information for rare and/or emerging species, such as Clavispora lusitaniae (n = 221), Saprochaete clavata (n = 184), Meyerozyma guilliermondii complex (n = 150), Candida haemulonii complex (n = 87), Rhodotorula mucilaginosa (n = 55), and Wickerhamomyces anomalus (n = 36).

Preliminary results were presented at the 28th European Congress of Clinical Microbiology and Infectious Diseases (21–24 April 2018, Madrid, Spain; abstract number 7965).; International audience; Isavuconazole is a recent extended-spectrum triazole with activity against yeasts. However, few data are available about the in vitro activity of rare yeast species. We report the MIC distribution of isavuconazole compared with fluconazole for a large collection of common or rare yeasts.METHODS:Isavuconazole and fluconazole MICs were determined using the EUCAST method for 1457 clinical isolates, mainly recovered from invasive infections, belonging to 29 species. They were sent to the National Reference Centre for Invasive Mycoses & Antifungals between January 2015 and October 2017 and species identification was performed using a polyphasic approach (matrix-assisted laser desorption/ionization time of flight analysis and a molecular method).RESULTS:Isavuconazole had effective in vitro activity against Cryptococcus neoformans (MIC90 < 0.25 mg/L), the five most common Candida spp. (MIC90 ≤ 0.5 mg/L for Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, and Candida krusei) and also against the majority of rare species, including Candida kefyr and Candida lusitaniae. A few isolates of C. albicans (0.7%, 3/404), C. glabrata (2.7%, 5/184), C. tropicalis (1.0%, 1/96) and C. parapsilosis (0.8%, 1/127) exhibited MIC ≥4 mg/L. All were also resistant to fluconazole according to the EUCAST breakpoints. Some isolates with isavuconazole MIC ≥4 mg/L were also observed among rarer species: Meyerozyma guilliermondii (8.7%, 2/23), Wickerhamomyces anomalus (10.0%, 1/10). Other rare species Saprochaete clavata, Magnusiomyces capitatus, and Rhodotorula mucilaginosa had high MIC50 (≥1 mg/L) and MIC90 (≥4 mg/L) and could be considered as resistant to isavuconazole.CONCLUSIONS:We confirmed the good in vitro activity of isavuconazole against common Candida, Cryptococcus species and the majority of the rare yeast species studied

International audience; Microsatellite length polymorphism (MLP) typing is a PCR-based method used for genotyping of the diploid yeast Candida albicans. However, MLP is subject to homoplasia which can hamper the accuracy of the results. We combined fragment length analysis, high-resolution DNA melting (HRM) analysis, and SNaPshot minisequencing after a single amplification of the CDC3 locus to study 95 epidemiologically independent C. albicans isolates. HRM analysis for a given electrophoretic group led to a maximum of three different curves due to the presence of a SNP upstream of the tandem repeat which could be characterized using the SNaPshot assay. The combination of the three methods had a discriminatory index of 0.88 in complete congruence with previous MLP typing (Mantel test R=0.99, P