Search

The main objective of this study was to identify carbapenem resistance mechanisms among clinical, commensal and environmental carbapenem-resistant Acinetobacter baumannii (CRAB) strains isolated in 5 Tunisian intensive care units (ICUs).CRAB isolates were recovered from different sources: clinical specimens, rectal and environmental swabs. Bacterial identification was carried out using conventional methods and susceptibility testing according to EUCAST recommendations. Evaluation of phenotypic carbapenemase production of was performed using seven different methods, and molecular detection of carbapenemase-coding genes (blaAll in all, 46 CRAB isolates were identified in clinical specimens (n = 26/26), rectal swabs (n = 17/36) and environmental swabs (n = 3/63). Most of them (n = 41/46) were clonally related and found in the different ICUs. All of the CARB isolates harbored blaThe emergence and diffusion of clonal CRAB strains inducing high mortality rates in ICUs is a major public health concern. Enhanced infection control practices and mandatory staff education are needed to control the spread of these multidrug-resistant bacteria.

Tuberculous Spondylodiscitis is the most common form of musculoskeletal tuberculosis. Molecular techniques on fresh tissues are proved to improve the diagnosis of tuberculous spondylodiscitis and to allow a rapid diagnosis to initiate the treatment and prevent neurological complications.The objective of the present study was to assess the diagnostic performances of single tube nested PCR and GeneXpert ultra in the diagnosis of tuberculous spondylodiscitis on formalin fixed paraffin embedded tissues.This study included 63 tuberculous spondylodiscitis cases collected from June 2014 to January 2020 and corresponding to 27 definite tuberculous spondylodiscitis with positive microbiology, and 36 probable tuberculous spondylodiscitis, with histopathological, clinical and radiological findings consistent with tuberculous spondylodiscitis but with negative microbiology. The sensitivity, specificity, positive predictive value and negative predictive value of nested PCR and GeneXpert ultra were determined with reference to microbiology.Nested PCR was positive in 47 (75%) cases: 26/27 definite tuberculous spondylodiscitis and 21/36 probable tuberculous spondylodiscitis. GeneXpert ultra was positive in only 6 (10%) cases corresponding to definite tuberculous spondylodiscitis. The sensitivity, specificity, positive predictive value and negative predictive value of nested PCR on formalin fixed paraffin embedded tissues were 96%, 100%, 100%, 83% respectively. For GeneXpert ultra, these rates were 22%, 100%, 100% and 25% respectively.Nested PCR and GeneXpert ultra on formalin fixed paraffin embedded tissues are useful tools for the diagnosis of tuberculous spondylodiscitis, especially for cases where microbiological investigations were not carried out. Both techniques have excellent specificity but single tube nested PCR is more sensitive. Key Points • Molecular techniques are routinely performed on fresh tissues • GeneXpert and nested PCR on formalin fixed paraffin embedded tissues are reliable for the diagnosis of tuberculous spondylodiscitis • Nested PCR is more sensitive than Genexpert for diagnosing tuberculous spondylodiscitis.

Objective: To determine the correlation between superficial, and intra-operative specimens in diabetic foot infections (DFIs).Methods: We conducted a cross-sectional study in patients with DFIs hospitalized in a Tunisian teaching hospital. Superficial specimens were collected for all patients, and intra-operative specimens were collected in operated patients. The specimens were processed using standard microbiology techniques. Antimicrobial susceptibility testing was carried out according to the protocol established by the European Committee on Anti-microbial Susceptibility Testing. Intra-operative and superficial specimens were considered correlated if they isolated the same microorganism(s), or if they were both negative.Results: One hundred twelve patients, 81 males and 31 females, mean age 56 years, were included. Superficial samples were positive in 77% of cases, and isolated 126 microorganisms. Among the positive samples, 71% were monomicrobial. The most frequently isolated microorganisms were Enterobacteriaceae (53%), followed by streptococci (21%) and Staphylococcus aureus (17%). Nine microorganisms (7%) were multi-drug resistant. Intra-operative samples were positive in 93% of cases. Superficial specimens were correlated to intra-operative specimens in 67% of cases. Initial antibiotic therapy was appropriate in 70% of cases. The lower-extremity amputation and the mortality rates were 41% and 1%, respectively.Conclusion: In our study, DFIs were most frequently caused by Enterobacteriaceae and superficial specimens were correlated to intra-operative specimens in only two thirds of cases. Clinicians should emphasize on the systematic practice of intraoperative specimens in all patients with DFIs treated surgically, while well-performed superficial specimens could be useful for prescribing appropriate antibiotic therapy in other patients.Keywords: Diabetic foot infections, superficial specimens, intra-operative specimens.

Emergence of the New Delhi metallo-β-lactamase (NDM-1), an Ambler class B metallo-β-lactamase able to hydrolyse all β-lactams except monobactams, constitutes a critical and increasingly important medical issue. The acquisition of blaNDM-1 is of particular concern for Proteus mirabilis, which is intrinsically resistant to tetracycline, tigecycline and colistin, as this will make clinical treatment extremely difficult. To the authors' knowledge, this is the first report of the blaNDM-1 gene in an extensively-drug-resistant P. mirabilis clinical isolate carrying plasmid-mediated resistance to carbapenems (blaNDM-1), cephalosporins (blaCMY-4), aminoglycosides (aph3 VIa and aph3 Ia) and fluoroquinolones (qnrA6).

The diagnosis of Tuberculous Cervical lymphadenitis (TCL) is challenging. The present study aimed to assess the performance of GeneXpert ultra (GXu) in the diagnosis of TCL on Formalin Fixed, Paraffin Embedded Tissues (FFPET). This study included 35 TCL cases confirmed by positive microbiology and/or positive GXu on Fresh Tissues (FT). The diagnostic performance parameters of GXu on FFPET were determined with reference to microbiology (positive Ziehl Neelsen and/or positive culture) and with reference to positive microbiology and/or positive GXu on FT. The GXu on FFPET was positive in 26/35 (74%) cases. With reference to positive ZN and or culture, the sensitivity, specificity, positive predictive value, and negative predictive value of GXu on FFPET were 63%, 100%, 100% and 71% respectively. With reference to positive microbiology and/or positive GXu on FT, these rates were 74%, 100%, 100% and 40% respectively. GXu on FFPET is a reliable tool for the detection of Mycobacterium tuberculosis complex particularly for cases where microbiological investigations have not been performed.

Objectives: Multidrug resistant Corynebacterium species are increasingly reported as the ethiological agent of various clinical infections. Thus, the purpose of this research was to evaluate the in vitro antimicrobial activity of three essential oils Salvia sclarea, Eucalyptus globulus and Eugenia caryophyllata against Corynebacterium species. Methods: Twenty-four multidrug resistant strains including C. striatum, C. amycolatum, C. urealyticum, C. aurimucosum, C. imitans, and C. jeikeium were used in the study. Inhibition diameter zone, minimum inhibitory concentration and minimum bactericide concentration of these oils were determined using agar disc diffusion method and microdilution method. Tigecycline was used as positive control. Results: Our study showed that Eugenia caryophyllata had the best activity. Eucalyptus globulus extract exhibited a moderate activity and Salvia sclarea was inactive against all the species tested. We found that C. amycolatum was more resistant to the essential oils than other species. On the other hand, tigecycline was effective on the majority of the strains (37.5%), but his action was lower than Eugenia caryophyllata oil. Conclusion: These results support the use of clove oil as a natural alternative to treat infections caused by multidrug resistant corynebacteria.

Resume L’examen cytobacteriologique des urines represente l’une des principales activites de routine d’un laboratoire de microbiologie. Compte tenu de cette charge de travail, l’analyse cytologique par methode microscopique manuelle est difficile a maitriser et manque parfois d’objectivite. L’automatisation de la cytologie urinaire permet de standardiser cet examen. Les performances de l’UriSed® (77 Electronica, Budapest, Hongrie) ont ete evaluees et les resultats de la numeration leucocytaire compares a la methode microscopique manuelle. La sensibilite (Se), la specificite (Sp), la valeur predictive positive (VPP) et negative (VPN) de l’automate etaient respectivement de : 85,1%, 88,7%, 85,1% et 88,7%. Une bonne correlation entre les deux methodes a ete notee, notamment pour les echantillons dont le taux de leucocytes est inferieur a la valeur seuil de 10 leucocytes/mm3. Dans 67% des cas d’urines a culture positive, les resultats de la bacteriurie determinee par l’UriSed® etaient concordants. En considerant une bacteriurie a 1 (+) comme seuil de pathogenicite, les Se, Sp, VPP et VPN etaient respectivement de 66,7%, 66,1%, 29,3% et 90,4% alors qu’avec un seuil de (++), les Se, Sp, VPP et VPN etaient de : 37,9%, 94,2%, 57,9% et 87,7%. Malgre le fait qu’il ne permette pas un gain de temps technique par rapport a la methode conventionnelle, l’UriSed® permet de standardiser et de maitriser l’analyse cytologique des urines avec une fiabilite globale acceptable.

Objectifs Les bacteries resistantes aux fluoroquinolones (FQs) ont ete frequemment identifiees comme cause de complications infectieuses apres biopsie de la prostate (BP). L’objectif de ce travail est d’etudier la prevalence du portage digestif d’enterobacteries resistantes aux FQs (ERFQ) chez les patients proposes pour BP et d’etudier l’interet de la prophylaxie ciblee selon la culture rectale sur le taux de complications infectieuses apres BP. Methodes Il s’agit d’une etude prospective qui a randomise 161 patients proposes pour BP en deux groupes. Groupe 1 (G1) : les patients ont recu une antibioprophylaxie empirique par la ciprofloxacine. Groupe 2 (G2) : un ecouvillonnage rectal a ete realise 10 jours avant la BP et ces patients ont recu une antibioprophylaxie adaptee selon la culture rectale. La prevalence du portage digestif d’ERFQ et les facteurs de risque de portage de germe resistants ont ete etudies. Une etude comparative de l’incidence des complications infectieuses apres la BP dans chaque groupe a ete realisee. Resultats Le G2 comprenait 84 contre 77 patients pour le G1. Il n’y avait pas de difference entre les deux groupes pour l’âge, le diabete, le volume de la prostate, le PSA, le nombre de carottes de biopsiques, et les facteurs de risque de resistance aux FQs ( Tableau 1 ). Dans le G2, la prevalence du portage digestif des ERFQ etait de 57,1 %. En cas de portage digestif des ERFQ, une antibioprophylaxie adaptee selon la culture rectale par cephalosporines de 3e generation a ete realisee dans 77 % des cas. Les patients ayant une ERFQ avaient un antecedent de prise de FQs au cours des six derniers mois dans 16,7 % des cas (p = 0,04) ( Tableau 2 ). Le taux de complications infectieuses apres BP etait de 13 % dans le G1 et 3,6 % dans le G2 (p = 0,02). Conclusion La prevalence du portage digestif d’ERFQ chez les patients proposes pour BP etait elevee (57,1 %). Le facteur de risque de resistance dans notre etude etait la prise de FQs au cours des six derniers mois. L’utilisation d’une antibioprophylaxie adaptee a la culture rectale, principalement par les cephalosporines de 3e generation, a considerablement abaisse nos taux de complications infectieuses apres BP.

Introduction La pyelonephrite aigue lithiasique (PNAL) est une urgence diagnostique et therapeutique frequente et potentiellement grave. Cette etude rapporte le profil bacteriologique et epidemiologique des PNAL au sein d’un service d’urologie. Materiels et methodes Etude retrospective realisee entre le 01/01/2019 et le 10/01/2020 incluant les patients admis pour PNAL confirmee. Tous les patients presentant une lithiase urinaire obstructive, associee a des lombalgies febriles ont recu une antibiotherapie probabiliste et un drainage des voies excretrices en urgence avec prelevement systematique des urines pyeliques (PUP) pour examen cytobacteriologique. Les hemocultures (HC) n’ont pas ete faites systematiquement. L’identification des bacteries isolees a partir d’un examen cytobacteriologique des urines (ECBU), des HC ou des PUP a ete effectuee selon les methodes conventionnelles, l’etude de la sensibilite aux antibiotiques par la methode de l’antibiogramme (normes du CA-SFM). Resultats Un total de 57 patients (18 hommes, 32 %) a ete inclus, dont 3 (5 %) se sont presentes dans un tableau de sepsis et 2 (3,5 %) dans un etat de choc septique. Un total de 35 bacteries ont ete isolees, 81 % a partir de L’ECBU, 4,5 % des HC et 47 % des PUP. L’ECBU etait negatif dans 30 cas (42 %) parmi lesquels 6 PUP etaient positifs, et aucune hemoculture positive. L’enquete bacteriologique a ete negative chez 9 patients. Tous les germes isoles etaient des enterobacteries, predominees par E. coli chez 21 patients (37 %), K. pneumoniae chez 7 patients (12 %), P. mirabilis chez 6 patients (11 %), et C. freundii chez 1 patient (2 %). La resistance aux cephalosporines de 3e generation a ete notee chez 11 souches (31 %). Quatre enterobacteries (11, 5 %) etaient productrices de betalactamase a spectre elargi (EBLSE). Deux souches etaient resistantes aux carbapenemes (1KP et 1 C. freundii). Concernant les aminosides, la resistance a la gentamicine a ete observee dans 4 cas (11,5 %) et l’amikacine etait efficace dans 91 % (32 souches). Sept souches (20 %) etaient resistantes aux fluoroquinolones et 10 (28,5 %) au cotrimoxazole. L’association cefotaxime et gentamicine a ete utilise comme traitement probabiliste chez 50 patients (88 %). L’evolution apres antibiotherapie adaptee et drainage des cavites excretrices a ete favorable chez 55 patients. Un seul patient, dont l’enquete bacteriologique etait negative, est decede dans un tableau d’etat de choc septique avec un syndrome de defaillance multiviscerale. Conclusion Le profil bacteriologique des PNAL de notre etude est concordant avec l’ecologie bacterienne des infections urinaires. Cependant, la proportion d’enterobacteries BLSE est tres importante, ce qui menace la reponse a l’antibiotherapie probabiliste utilisee selon les recommandations actuelles.

Objectives: In this study we investigated the susceptibility profiles against macrolides and lincosamides of 85 C. striatum strains isolated at a clinical centre in Sousse (Tunisia). Methods: The strains were identified by the routine biochemical assays and then confirmed by Vitek-Maldi-Tof-MS. MIC?s of erythromycin and clindamycin were determined using the microdilution method. The detection of erm(X), erm(B), msr(A), mph(A) and mef(A-E) resistance genes was performed by PCR. The strains were typed by PFGE using XbaI. Results: Sixty-nine (81.17%) strains were resistant to erythromycin, 58 (67.44%) strains were resistant to clindamycin. There was a high correlation between the resistance to erythromycin and clindamycin and the presence of erm(X) gene in 85.50% and 89.65% respectively. The erm(B) gene was detected in 21(24.70%) strains whereas, no others genes were detected in our strains collection. By PFGE, the 85 strains belonged to 18 different clones. Conclusion: erm(X) is implicated in macrolide resistance for almost all the Corynebacterium strains analyzed in our study. Other resistance genes like erm(B) must also be implicated in this resistance, although its presence seems to be unusual in previously reported studies.

Corynebacterium spp. are part of the human microbiota. Recently, species of this genus are increasingly implicated in different types of infections especially in immunocompromized and hospitalized patients. The significance of the presence of the genus Corynebacterium in foods is not clearly established. These bacteria may be involved in spoilage or ripening of cheese and meats. This review focused on different researches concerning the implication of Corynebacterium species in food’s contamination.

The aim of this study is to test a pooling approach for the RT-PCR test to detect low viral loads of SARS-CoV-2. We found that a single positive specimen can still be detected in pools of up to 10. Each laboratory should conduct its own evaluation and validation of pooling protocols according to its specific context., Highlights • Pooling samples of SARS-COV-2 is a cost-saving method with the ability to detect the virus in specimens with low viral load • We found that a single positive specimen can still be detected in pools of up to 10 • Additional amplification cycles, or inclusion of a re-test for negative group samples, could reduce false negative rate

Introduction Le diagnostic de la COVID-19 se base sur la detection d’une ou de deux regions specifiques du genome de SARS-CoV-2 par RT-PCR en temps reel. Le Laboratoire national de reference (LNR) de surveillance de la grippe et des autres viroses respiratoires a ete designe pour assurer cette tâche jusqu’au 27 mars, date a laquelle le reseau des laboratoires a ete elargi pour augmenter la capacite nationale du testing. La presente etude rapporte l’experience du LNR pendant la periode ou il a assure a lui seul le diagnostic de la COVID-19 (6 fevrier au 26 mars). Materiels et methodes Apres optimisation de la technique de RT-PCR en temps reel se basant sur le protocole de Hong Kong (genes cibles N et ORF1b), l’analyse de 1913 prelevements nasopharynges a ete realisee. Les resultats etaient quotidiennement adresses a l’Observatoire national des maladies nouvelles et emergentes (ONMNE) pour assurer la surveillance, l’investigation et le contact tracing en cas de detection de cas positifs et ce afin de limiter l’extension de la maladie. Resultats Au cours de cette periode, 229 (12 %) cas positifs ont ete colliges. Le 2 mars, le premier cas a ete diagnostique. Il s’agissait d’un cas importe d’Italie. Ce dernier a ete place en confinement. Le 10 mars, les deux premiers cas autochtones ont ete identifies. La capitale et les villes avoisinantes etaient les regions les plus touchees avec 113 (49,3 %) cas. Le sex-ratio etait de 1,14 soit 122 hommes (53,3 %) et 107 de femmes (46,7 %). La moyenne d’âge etait de 46,18 ans avec des extremes allant de 6 mois a 85 ans. La tranche d’âge entre 15 et 45 ans etait la plus touchee avec une frequence de 48,9 %. La transmission etait autochtone chez 117 (51,1 %) patients, versus 112 (48 %) cas importes. La majorite des cas etaient symptomatiques (82,5 %). La fievre (93 %), la toux (77 %) et l’asthenie (43 %) etaient les symptomes les plus rapportes. Les cas symptomatiques etaient significativement plus frequents parmi les cas importes 93 % contre 72,6 % des cas autochtones avec un OR de 4,9 et un intervalle de confiance 95 % = [2,14–11,23]. Les mesures prises pour les malades asymptomatiques etaient l’auto-isolement a domicile (64,2 %) ou l’hebergement dans des centres de confinement (16,6 %). Quarante quatre (19,2 %) patients avaient necessite une hospitalisation dont 20 (43,5 %) en reanimation avec un taux de letalite de 7,4 % (17 deces sur 229). Le personnel de sante representait 12,2 % (28 cas) du total des personnes infectees par le SARS-CoV-2, avec une transmission majoritairement autochtone (75 %). Conclusion Les mesures judicieuses entreprises au cours de cette periode ont permis d’etablir un plan de prevention et de riposte au risque de diffusion du SARS-CoV-2. Ceci est etroitement lie a la collaboration entre les differents intervenants de tous les secteurs.

Introduction La prevalence des infections associees aux soins a enterobacteries productrices de carbapenemases (EPC) est en augmentation dans le monde. Ces infections, difficiles a traiter et souvent source d’impasses therapeutiques, representent un danger pour les patients avec un risque d’epidemies hospitalieres pouvant devenir incontrolables. Materiels et methodes Une etude retrospective a ete menee entre janvier 2015 et decembre 2019, portant sur les patients qui ont ete hospitalises au service d’urologie chez qui une EPC a ete identifiee sur l’examen cytobacteriologique des urines (ECBU). Les caracteristiques epidemiologiques, cliniques et bacteriologiques de ces infections ont ete etudiees. Resultats Au cours de la periode d’etude, nous avons identifie 25 patients porteurs d’EPC. L’âge moyen etait de 50,5 ans. Une hospitalisation dans les 3 mois precedents a ete retrouvee dans 11 cas (44 %), la prise d’antibiotiques dans les 6 mois precedant l’infection a ete retrouvee dans 13 cas (52 %), et 11 patients (44 %) avaient un ECBU positif a un autre germe dans les 12 mois precedant l’infection. Au cours de l’hospitalisation, un geste urologique a ete realise dans 20 cas (80 %), un materiel etranger urologique etait en place dans 15 cas (60 %) et le delai moyen entre l’hospitalisation et l’infection etait de 12 jours ± 4,8. Sur le plan bacteriologique, les souches identifiees etaient K. pneumoniae (n = 20), E. cloacae (n = 2), E. coli (n = 2) et R. terrigena (n = 1). L’etude de la sensibilite aux antibiotiques a montre qu’elles etaient toutes resistantes a l’ertapeneme, que 10 l’etaient a l’imipeneme (40 %) et que 13 l’etaient au meropeneme (52 %). Par ailleurs, toutes les souches etaient resistantes aux cephalosporines de 3e generation et sept l’etaient a l’amikacine (28 %). Une souche etait resistante a la colistine (etudiee dans 18 cas), et quatre souches l’etaient a la tigecycline (etudiee dans 21 cas). Cliniquement, les EPC etaient responsables d’une bacteriurie asymptomatique dans 14 cas (56 %), et d’une infection declaree dans 11 cas (44 %), le plus souvent une pyelonephrite aigue (5 cas). Deux patients ont presente un choc septique dont un est decede des suites de l’infection. En cas d’infection declaree (11 cas), une double antibiotherapie adaptee a ete prescrite, associant colistine/amikacine(cinq cas), colistine/imipeneme (quatre cas) et imipeneme/amikacine (deux cas). Conclusion Les EPC representent une menace reelle et preoccupante car elles sont capables de provoquer des infections contre lesquelles les moyens therapeutiques sont extremement limites, de se disseminer rapidement et possedent un potentiel epidemiogene important. Les bonnes pratiques d’hygiene hospitaliere sont une pierre angulaire du controle de ces bacteries associee a une utilisation reflechie et limitee des carbapenemes pour diminuer la pression de selection des souches productrices de carbapenemases.

The pandemic spread of multidrug-resistant (MDR) bacteria (i.e., methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum b-lactamase-producing Enterobacteriaceae (ESBLPE), vancomycin-resistant enterococci, carbapenemase-producing Enterobacteriaceae (CPE), multiresistant Pseudomonas aeruginosa and multiresistant Acinetobacter baumannii ) pose a threat to healthcare Worldwide. We found limited data of MDR bacteria in pediatric patients hospitalized in Tunisian tertiary healthcare.The aim of the study is to evaluate the acquisition rate of MDR acquisition during hospitalization and to explore some of the associated risk factors for both carriage and acquisition at the pediatric department, Sahloul University Hospital. During September and October 2016, newly admitted patients were screened, at admission, during care and at discharge. Risk factors for colonization were explored by multivariate analysis. Of 112 newly admitted patients, 8.92% were colonized with at least one MDR. No risk factor was identified at admission. During hospitalization, five newly acquisition MDR (4.9%) were detected and eight (7.84%) at discharge. The specie most frequently detected on admission was Escherichia coli (50%), whereas, on discharge, Escherichia coli and K. pneumoniae were the species most frequently detected (52.7%). The pediatric intensive care unit (PICU) hospitalization, the length of hospital stay (more than 3days) and age under 2 years were identified as risk factor for acquisition of MDR during hospitalization. We identified several independent risk factors for contracting MDR bacteria during hospitalization in a tertiary pediatric department. The incidence of symptomatic MDR Infection among those colonized should be under close surveillance and long-term screening for those children is required. An institutional screening program for MDR especially in PICU might be discussed in regards to cost effectiveness. Keywords: Children; colonization; multidrug resistance; hospitalization

Emergence of the New Delhi metallo-β-lactamase (NDM-1), an Ambler class B metallo-β-lactamase able to hydrolyse all β-lactams except monobactams, constitutes a critical and increasingly important medical issue. The acquisition of bla

Resume Introduction Nous presentons les donnees concernant le profil bacteriologique et la sensibilite aux antibiotiques des bacteries urinaires isolees chez des enfants de la region de Sousse (Tunisie). Population et methodes Il s’est agi d’une etude retrospective portant sur les examens cytobacteriologiques d’urines positifs provenant d’enfants ou de nouveau-nes et parvenus a notre laboratoire durant les annees 2012 et 2013. L’identification bacterienne a ete realisee selon les methodes conventionnelles. La realisation et l’interpretation de l’antibiogramme ont ete effectuees selon les recommandations du comite de l’antibiogramme de la Societe francaise de microbiologie. Resultats Nous avons collecte 1879 bacteries non redondantes dominees a plus de 73 % par Escherichia coli . Les bacteries provenaient majoritairement d’enfants et de nourrissons (84 %) ; la plupart (65,7 %) provenaient de milieu communautaire. Le taux de resistance des enterobacteries aux beta-lactamines etait eleve, avec production de beta-lactamase a spectre etendu pour 12,8 % de celles provenant d’enfants vs 22,6 % des souches neonatales. Une frequente efficacite des furadoines, de l’imipeneme, de la fosfomycine et de la colistine a ete notee. Conclusion L’impact du large usage des beta-lactamines en milieux neonatal et pediatrique est mesurable. La colistine, l’imipeneme et la fosfomycine sont les antibiotiques les plus frequemment actifs sur les bacteries responsables d’infections urinaires pediatriques et neonatales ; toutefois, leur prescription ne peut se faire de maniere probabiliste. Les nitrofuranes, actifs sur les bacteries uropathogenes chez l’enfant et le nouveau-ne, presentent certaines limites d’utilisation sur ce terrain, et leur indication en cas de pyelonephrite reste a discuter.

Corynebacterium striatum is a nosocomial opportunistic pathogen increasingly associated with a wide range of human infections and is often resistant to several antibiotics. We investigated the susceptibility of 63 C. striatum isolated at the Farhat-Hached hospital, Sousse (Tunisia), during the period 2011–2014, to a panel of 16 compounds belonging to the main clinically relevant classes of antimicrobial agents. All strains were susceptible to vancomycin, linezolid, and daptomycin. Amikacin and gentamicin also showed good activity (MICs90 = 1 and 2 mg/L, respectively). High rates of resistance to penicillin (82.5%), clindamycin (79.4%), cefotaxime (60.3%), erythromycin (47.6%), ciprofloxacin (36.5%), moxifloxacin (34.9%), and rifampicin (25.4%) were observed. Fifty-nine (93.7%) out of the 63 isolates showed resistance to at least one compound and 31 (49.2%) were multidrug-resistant. Twenty-nine resistance profiles were distinguished among the 59 resistant C. striatum. Most of the strains resistant to fluoroquinolones showed a double mutation leading to an amino acid change in positions 87 and 91 in the quinolone resistance-determining region of the gyrA gene. The 52 strains resistant to penicillin were positive for the gene bla, encoding a class A β-lactamase. Twenty-two PFGE patterns were identified among the 63 C. striatum, indicating that some clones have spread within the hospital.

Summary Background Escherichia coli is the predominant pathogen causing urinary tract infection (UTI), the most common bacterial infectious disease encountered in clinical practice, accounting for significant morbidity and high medical costs. The severity of UTI produced by E. coli is due to the expression of a wide spectrum of virulence factors. In this study we evaluated the role of E. coli virulence determinants in the pathogenesis of UTI. Methods A total of 90 uropathogenic E. coli strains were screened by PCR for the prevalence of seven virulence genes encoding type 1 fimbriae ( fimH ), pili associated with pyelonephritis ( pap ), S and F1C fimbriae ( sfa and foc ), afimbrial adhesins ( afa ), cytotoxic necrotizing factor ( cnf ), hemolysin ( hly ), and aerobactin ( aer ). Results The prevalence of genes coding for fimbrial adhesive systems was 68% for fimH , 41% for pap , and 34% for sfa/foc . The operons coding for afa afimbrial adhesins were identified in 20% of strains. The hly and cnf genes coding for toxins were amplified in 19% and 3% of strains, respectively. A prevalence of 52% was found for the aer gene. The various combinations of detected genes were designated as virulence patterns. The strains isolated from hospitalized patients displayed a great diversity of gene associations compared to those isolated from ambulatory patients. Conclusions Our study showed that investigation of the bacterial pathogenicity associated with UTI may contribute to a better medical intervention.

Resume Objectif L’epidemiologie de l’infection par le virus herpes humain de type 8 (HHV8) est inconnue en Tunisie. Notre objectif est d’evaluer sa prevalence dans une population du Centre-Est tunisien, ainsi que chez des sujets a risque eleve de transmission virale sanguine ou sexuelle. Methodes Un effectif de 553 personnes a ete concerne : 116 donneurs de sang, 100 femmes enceintes, 100 enfants, 50 sujets ayant une infection sexuellement transmissible non infectes par le VIH et 50 autres infectes par le VIH, 107 sujets polytransfuses et 30 personnes ayant eu une greffe de rein. La technique de detection des anticorps anti-HHV8 etait l’immunofluorescence indirecte. Resultats La seroprevalence des IgG anti-HHV8 etait de 13,8 % chez les donneurs de sang, 13 % chez les femmes enceintes et 12 % chez les enfants. Dans la population saine adulte, il n’existait pas de difference de prevalence selon le sexe, les caracteristiques sociodemographiques, les antecedents chirurgicaux et les marqueurs serologiques d’hepatite B. La prevalence des IgG anti-HHV8 etait significativement elevee dans le groupe a risque sexuel (p Conclusion La prevalence du HHV8 est elevee dans le Centre-Est tunisien a l’instar des pays mediterraneens. L’infection parait s’acquerir tot dans l’enfance, probablement par voie salivaire. Le risque de transmission sanguine du virus, controverse dans la litterature, est clairement etabli dans cette etude effectuee en region endemique. Un depistage precoce de l’infection dans les populations a risque de sarcome de Kaposi parait necessaire dans nos regions.

Corynebacterium striatum is frequently encountered in the routine clinical microbiology laboratory. It is widely disseminated in the environment and constitutes part of the normal micro-biota of the skin and mucous membrane. Identification of this species by biochemical methods remains difficult and several misidentifications have been reported previously. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for the identification of this microorganism was designed based on the hypervariable region of the polymorphic RNA polymerase β-subunit gene (rpoB). All available Corynebacterium rpoB sequences were analyzed by computerassisted restriction analysis. The rpoB PCR-RFLP pattern predicted by using endonucleases MseI and NlaIV clearly differentiated C. striatum from all other Corynebacterium species. This method was successfully applied for the reliable identification of 67 C. striatum clinical isolates and can be used for the timely detection of infected patients or for epidemiological studies.

The objective of the study is to compare the performance of conventional fluorescence microscopy (CFM) and light-emitting diode (LED) fluorescence microscopy (FM) for detection of acid-fast bacilli (AFB) in clinical samples. We included AFB smears, stained using the auramine O method and blindly examined with both CFM and LED-FM. Culture results were used as reference for evaluating the reliability of the FM. We included 180 culture positive specimens and an equal number of culture negative specimens. Sensitivities for the CFM and LED-FM were 79.4% and 82.2%, respectively. Both microscopes had a high specificity (97.2%). The negative-positive (>1 cross) inter-reader agreement of LED-FM and CFM was excellent. Therefore, detection of scanty AFB was higher with LED-FM. Both microscopes were equivalent with respect to time required to read smears. Although it was not faster than CFM, the higher detection of scanty AFB smears combined with ease of use supports the consideration of LED microscopy by all tuberculosis diagnostic laboratories, as a replacement for conventional fluorescence microscopes.

Objectives Extended-spectrum β-lactamases are widespread in hospital settings worldwide. The prevalence of ESBL producing Klebsiella pneumoniae (ESBL-KP) strains isolated from patients has recently increased in Tunisia. We conducted this study to determine the prevalence and the genetic characterization of ESBL-KP in a Tunisian Hospital. Patients and methods We performed antibiotic susceptibility testing, multiplex PCR, and DNA sequencing analysis on 118 non repetitive K. pneumonia strains isolated during three years, to determine the prevalence and genotypes of ESBL among K. pneumoniae clinical isolates. Results Most ESBL-producing K. pneumonia strains were isolated from hospitalized patients, especially in neonatal and pediatric wards. The resistance to other antibiotics was high. Most of the pathogens were isolated from the urinary tract (86.44%). Carbapenems were the most effective antimicrobial agents followed by amikacin and fosfomycin. The rate of blaSHV, blaTEM, and blaCTX-M genes among the isolates was 89, 56.78, and 81.35%, respectively. Sequencing revealed the amplicons encoding TEM-1, TEM-53, TEM-158, SHV-1, SHV-11, SHV-28, CTX-M-15, CTX-M-15-like. The blaCTX-M-15 was the dominant gene among Tunisian isolates, but this was the first report of blaTEM-53 and blaTEM-158 genes in the country. Conclusions Our results confirm the predominance of CTX-M-15 in Tunisia. To our knowledge, this is the first report of TEM-158 and TEM-53 in Tunisia.

TB diagnosis and resistance detection remain to suffer from complex techniques and long time taking. The rapid identification of drug-resistant strains of Mycobacterium tuberculosis is crucial for the timely initiation of appropriate antituberculosis therapy. Over the years, several new technologies have been proposed to accelerate and simplify the detection of resistant M. tuberculosis. In this context, we evaluated the Genotype MTBDRplus to detect resistance to isoniazid and rifampin in positive M. tuberculosis cultures. The performance of the Genotype MTBDRplus assay was compared with that of the phenotypic drug susceptibility tests, a gold standard culture based method. The Genotype MTBDRplus assay was quicker and more cost-effective for the detection of rifampin and isoniazid resistance, with a slightly lower detection of rifampin-resistant strains in our setting.

Lactococcus lactis cremoris is rarely involved in human pathology. A thirty two-year old pregnant woman with premature rupture of membrane history presented with chorioamnionitis due to L. lactis cremoris. She underwent an emergency caesarian section and was treated with antibiotics including the association of amoxicillin and clavulanic acid. She was completely recovered. This is the first case to our knowledge of chorioamnionitis due to this organism., Highlights • Lactococcus lactis cremoris is rarely involved in human pathology. • A thirty two-year-old pregnant woman had premature rupture of membrane history. • She presented with chorioamnionitis due to Lactococcus lactis cremoris. • She underwent an emergency caesarian section and was treated with antibiotics. • This is the first case to our knowledge of chorioamnionitis due to this germ.

Introduction: Salmonella Livingstone is one of the most common serotypes responsible for nosocomial outbreaks in Tunisia. In this study, 42 isolates of Salmonella Livingstone were analyzed. Most of these were isolated from humans (31 strains from Tunisia and 9 strains from Belgium) and 2 isolates came from food products (beef and pork). Methodology: All strains were characterized by antibiogram, multilocus sequence typing (MLST), and virulotyping. This last technique was carried out by simple PCR of five chromosomal genes (agfA, hin/H2, iroB, phoP/Q, and slyA) and two plasmid genes (spvA and spvC). Results: All Tunisian strains were resistant to amoxicillin, amoxicillin-clavulanic acid, ticarcillin, cefalotin, gentamicin, and kanamycin. They were also resistant to third-generation cephalosporin antibiotics (cefotaxim and ceftazidim). Belgian isolates were susceptible to all antibiotics tested. Further to MLST analyses, Tunisian strains belonged to the same sequence type, ST543. For Belgian isolates, eight strains had a ST543 profile, two strains had a ST638 profile, and one strain had a ST457 profile. Analyses of the virulence gene contents showed that strains isolated in different years and from different origins had the same virulence profile. These carried all five chromosomal genes and lacked plasmid-located virulence genes spvA and spvC. Conclusions: A combination of different typing methods showed that the majority of Belgian strains and all Tunisian strains were closely related; they belonged to the same sequence type (ST543) and had the same virulence profile, but different antibiotic resistance profiles depended on the country of origin.

We evaluated the performance and the cost of chromogenic medium Uriselect4 agar with regard to the standard medium for the detection and identification of urinary tract pathogens. A total of 503 clinical urine specimens containing leucocytes greater or equal to 104/mL were analysed prospectively, in parallel by two different persons on blood agar (GS) and Uriselect4 according to the manufacturers' instructions. Of the 503 urine specimens tested, 210 gave a positive culture on Uriselect4 versus 181 on GS. The majority of bacterial species grew on both media; enterobacteria grew on Uriselect4 better than GS. The identification of Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis), KES group and Enterococcus faecalis (E. faecalis) did not require the use of galleries Api and has a gain of 24 h. Positive pure cultures on Uriselect4 corresponding to negative cultures of GS were noted in 17 ases. Conversely, in seven cases a positive pure culture on GS was noted while the corresponding Uriselect4 cultures were negative. The cost of identification on GS (including the cost of galleries Api), was about two times higher than Uriselect4. Uriselect4 medium isolates the most frequent urinary tract pathogens and identify them so almost immediately, with a lower cost.

Background In Tunisia, country of intermediate endemicity for Hepatitis B virus (HBV) infection, most molecular studies on the virus have been carried out in the North of the country and little is known about other regions. The aim of this study was to determine HBV genotype and subgenotypes in Central-East Tunisia. A total of 217 HBs antigen positive patients were enrolled and determination of genotype was investigated in 130 patients with detectable HBV DNA. HBV genotyping methods were: PCR-RFLP on the pre-S region, a PCR using type-specific primers in the S region (TSP-PCR) and partial sequencing in the pre-S region. Results Three genotypes (D, B and A) were detected by the PCR-RFLP method and two (D and A) with the TSP-PCR method, the concordance between the two methods was 93%. Sequencing and phylogenetic analysis of 32 strains, retrieved the same genotype (D and A) for samples with concordant results and genotype D for samples with discordant results. The sequences of discordant genotypes had a restriction site in the pre-S gene which led to erroneous result by the PCR-RFLP method. Thus, prevalence of genotype D and A was 96% and 4%, respectively. Phylogenetic analysis showed the predominance of two subgenotypes D1 (55%) and D7 (41%). Only one strain clustered with D3 subgenotype (3%). Conclusions Predominance of subgenotype D7 appears to occur in northern regions of Africa with transition to subgenotype D1 in the East of the continent. HBV genetic variability may lead to wrong results in rapid genotyping methods and sequence analysis is needed to clarify atypical results.

The pathogenic capacity of streptococcus agalactiae or Group B Streptococcus (GBS) has been increasing in nonpregnant adults. We carried out a study about the pathogenic capacity and susceptibility to antibiotics of GBS strains isolated between January 2003 and June 2004 in Farhat Hached hospital of Sousse (Tunisian general hospital).Identification of strains was carried out according to usual bacteriological characters. Susceptibility of GBS to antibiotics was carried out according to CA-SFM recommendations.during the study period, sixty strains were isolated. They were essentially isolated from the urinary tract (52%) and the skin (36.2%). Women accounted for 68 percent of the cases. Among the 30 hospitalized patients, 20 (66%) had serious underlying disease. Diabetes mellitus was predominant (80%). All group B streptococci isolates were susceptible to amoxicillin, penicillin G, vancomycin and to pristinamycin. Sixty four percent of isolates were susceptible to erythromycin. Among the invasive infection, caused by GBS in nonpregnant adults, skin and urinary tract infections were the most common. All isolates were susceptible to amoxicillin, penicillin G. vancomycin and to pristinamycin.