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Ventilator-associated pneumonia (VAP) represents a major cause of nosocomial infections in the intensive care units in which Staphylococcus aureus is frequently involved. Better knowledge of this pathogen is required in order to enhance the patient's treatment and care. In this article, we studied the bacteriological profile and virulence factors of S. aureus-related VAP on a 3-year period. We included a collection of S. aureus strains (n = 35) isolated from respiratory samples from patients diagnosed with VAP in the intensive care units. We studied the bacteriological aspects and we searched for the presence of virulence factors (SpA, FnbpA, Hla, and PVL genes) in the strains, and we also studied the clinical and biological aspects of the infections. The average age of our patients was of 36 years and they were predominantly males (sex ratio = 3.37). A severe head trauma or a history of coma was noted in 73.43% of the patients. The average duration of ventilation was 29 days. Among the studied strains, five were Methicillin-resistant S. aureus of which three expressed the mecA gene. Overall, the Hla gene was detected in 85.7% of the strains and it was more prevalent in Methicillin-susceptible than Methicillin-resistant strains (93.3% versus 40%; P = 0.014). FnbpA, Spa, and PVL genes were detected, respectively, in 80%, 45.7%, and 20% of the strains. Therefore, our studied strains were essentially associated with the production of Hla and FnbpA genes. It is, however, important to elucidate their expression in order to establish their role in the VAP pathogenesis.

Peritonitis is a major complication of peritoneal dialysis (PD) and due to its gravity, it remains the primary reason to switch from PD to hemodialysis. Elizabethkingia meningoseptica, a non-fermentative Gram-negative bacillus, is rarely encountered as a pathogen causing peritonitis in adults. We present here a case report of an acquired infection with this organism in adult on PD. To the best of our knowledge, this is the first report of infection with this organism in a continuous ambulatory PD patient in Tunisia.

Introduction. Group A Rotavirus (RVA) is known to be a major cause of acute gastroenteritis (AGE) in children but its role as a potential pathogen in immunocompetent adults is probably underestimated. Aim. To compare RVA infections in patients from different age groups. Methodology. Fecal samples were collected from patients aged from birth to 65 years, hospitalized or consulting for AGE between 2015 and 2017. All samples were screened by RT-PCR for the detection of VP6 gene specific of RVA. RVA-positive samples were VP7 and VP4 genotyped using multiplex semi-nested RT-PCR. Full-length VP7 gene of G9-positive strains were sequenced and submitted for phylogenetic analysis. Results. Of 1371 stool specimens collected from children (<5 years; n =454), older children (5 to <15 years; n =316) and adults (15-65 years; n =601), 165 (12.0 %) were RVA-positive. RVA detection rates were significantly higher in children and adults than in older children (15.8 % and 12.1 Vs 6.3 %, respectively; P <0.001). While RVA infections were mostly detected during the coldest months in children, they were observed all year-round in patients aged >5 years. Although G1P[8] remained the most prevalent combination (41.7 %) detected in children, G9P[8] strains widely predominated in adults (58.1 %), followed by G2P[4] (12.9 %). All characterized G9 strains clustered in the modern lineage III. Conclusion. RVA play an important role in AGE not only in children but also in adults. The findings of a wide G9 predominance in patients >5 years highlights the need for continuing surveillance in both pediatric and mature populations.

Introduction. Tunisia is an intermediate hepatitis B virus (HBV) endemic country. The vaccination against hepatitis B was introduced in 1995 including four doses with a first dose administrated at birth. Decreasing the level of antibodies against hepatitis B surface antigen (anti-HBs) over time can be alarming. This study was conducted to explore the anti-HBV immune response among children under 6 years old, vaccinated according to the national vaccination schedule, by evaluating the immunological response to primary vaccination and by exploring the anamnestic immune response to a booster dose. Methods. We conducted a cross-sectional prospective study from June 2016 to June 2017 ( n =180), based on voluntary participation. Children were recruited from the public pediatric ward sectors in Sahloul University Hospital of Sousse in Central Tunisia. An anti-HB titre was determined based on electro-chemiluminescence micro-particle immunoassay (ECLIA), using Elecsys Anti-HBs II kit, Roche. Results. Mean age at the time of enrollment in the study was 33±14.8 months. The seroprotection rate was 77.2 %. The anti-HB titre differed significantly between the different age groups ( P =0.002). The predicting variable for having no seroprotective antibody level was older age. Children with anti-HB levels <10 IU l - 1 were offered an additional dose of HBV vaccine. Anamnestic response 1 month after the challenge dose was observed in 100 % of subjects. The probability of developing a high antibody response, following the booster dose increased in conjunction with an increased pre-booster antibody level. Conclusion. The response to a booster dose suggests the persistence of immune memory in almost all vaccinated individuals. Although a booster dose increases substantially anti-HB titre, the clinical relevance of such an increase remains unknown.

The aim of the present study was to report the molecular characterization of human group A rotaviruses (RVAs) circulating in Tunisia. Stool specimens were collected from children under 5 years of age who had been hospitalized or were consulting for gastroenteritis in Tunisian hospitals between 2015 and 2017. All samples were screened by reverse-transcription polymerase chain reaction (RT-PCR) for the detection of the VP6 gene specific for RVA. RVA-positive samples were further analysed for G/P genotyping by semi-nested multiplex RT-PCR. Among 454 tested samples, 72 (15.8 %) were positive for RVA. G1P[8] was the most prevalent detected strain (41.7%), followed by G9P[8] (32.8%), G2P[4] (7.5%), G12P[8] (7.5%), G1P[6] (3.0%), G2P[8] (1.5%) and G3P[8] (1.5%), with mixed infections in 4.5 % of cases. In the absence of a national anti-rotavirus vaccination strategy, RVAs remain the primary aetiological agent for gastroenteritis in Tunisian children.

Objectives: The whole-genome sequence (WGS) of Klebsiella pneumoniae KP3771 isolate was characterized. This strain was recovered from the urine sample of an 80-year-old man hospitalized in an intensive care unit of the University Hospital Tahar Sfar in Tunisia. Materials and Methods: WGS using a MiSeq platform was used. The assembled genome was subjected to several software analyses. Results: K. pneumoniae KP3771 was resistant to all antibiotics but colistin and tigecycline. WGS analysis found 18 transmissible genes encoding resistance markers, including bla NDM-1 and bla CTX-M-15 genes, which were carried by four plasmids belonging to the Inc Ib, IIk, and R groups. Three families of genes encoding virulence factors were detected, including adhesins ( fimH , fimA , fimB , fimC , mrkD , Kpn , and ycfM ), siderophores (enterobactin, aerobactin, and yersiniabactin siderophores), and protectin/invasin ( traT ). The strain was assigned to the sequence type 147. Conclusions: This study describes the genome of a carbapenemase-producing K. pneumoniae clinical isolate recovered in Tunisia. Bacteria WGS has become the reference technology to address epidemiological issues; this high level of information is particularly well suited to enrich epidemiological workflows' output.

Background: New Delhi metallo-β-lactamase (NDM)-producing Acinetobacter baumannii have been described in several countries worldwide, and studies have suggested that Acinetobacter spp. could play the role of intermediate progenitor of the bla NDM-1 gene between environmental progenitor and Enterobacteriaceae., Materials and Methods: In total, 246 carbapenem-resistant A. baumannii isolates from a teaching hospital in Sousse, Tunisia were investigated between 1 st June 2013 and 31 st December 2015 to detect metallo-ß-lactamase (MBL) production. Polymerase chain reaction (PCR), antibiotic susceptibility testing, and genetic and whole-genome sequencing tools were used to study the underlying carbapenem resistance mechanisms., Results: PCR screening of the 246 carbapenem-resistant A. baumannii isolates revealed that 242 of 246 isolates harboured carbapenemase genes (seven of 246 positive for bla NDM-1 , four of 246 positive for bla NDM-1 and bla OXA-23 , 231 positive for bla OXA-23 ). Conjugation and electroporation experiments suggested that the bla NDM-1 gene is likely to be chromosomally located. All the NDM-1-producing A. baumannii isolates were clonally related, and belonged to ST85 according to the Pasteur Institute's multi-locus sequence typing scheme. Analysis of the immediate genetic environment of the bla NDM-1 gene revealed that the gene was located within a truncated isoform of Tn125 transposon (ΔTn125). The bla OXA-23 gene was located within transposon Tn2008., Conclusion: This study showed the dissemination of a single clone of NDM-1-producing A. baumannii in a Tunisian hospital. Countries in north Africa may constitute a significant reservoir for NDM-1-producing A. baumannii. The spread of the bla NDM-1 gene in A. baumannii was linked to clonal spread in this study., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Acinetobacter baumannii is an opportunistic pathogen in healthcare facilities responsible for nosocomial infections mostly in immunocompromised patients. Colistin resistance is increasingly reported worldwide in A. baumannii. Here we describe the in vivo selection of colistin and rifampicin resistance in carbapenem-resistant A. baumannii. Antimicrobial susceptibility testing, plasmid analysis and whole-genome sequencing (WGS) were performed to fully characterise the resistome of two clinical isolates (AbS1 and AbS2) selected during treatment. Clinical isolate AbS1 remained susceptible to colistin, rifampicin and tigecycline, whilst AbS2 was susceptible only to tigecycline. PCR analysis revealed the presence of a bla OXA-23-like carbapenemase gene. Kieser extraction revealed an ca. 74 kb plasmid harbouring bla OXA-23 . WGS revealed genomes of 3.8 Mbp in size with a G + C content of 38.9%, and both belonged to ST281 according to the Oxford MLST scheme and ST641 according to the Institut Pasteur scheme. The resistome was also composed of naturally occurring β-lactamases, i.e. ADC-25 cephalosporinase and OXA-82 oxacillinase, aminoglycoside resistance genes [aac(3)-Ia, aadA1 and aph(3')-VIa (aphA6)], and mutations in DNA gyrases explaining fluoroquinolone resistance. Single nucleotide polymorphism analysis revealed that both isolates were identical except for a 30-nucleotide duplication within the pmrB gene and a point mutation in the rpoB gene resulting in colistin and rifampicin resistance, respectively. This study highlights the genomic plasticity of A. baumannii under antibiotic pressure. The 10-amino acid duplication in PmrB affects colistin susceptibility by regulating lipopolysaccharide modification through the PmrAB two-component system. These findings provide further information on the molecular mechanisms leading to colistin resistance in A. baumannii., (Copyright © 2017 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.)

The emergence of colistin-resistant Klebsiella pneumoniae (CoR Kp ) is a public health concern, since this antibiotic has become the last line of treatment for infections caused by multidrug-resistant (MDR) Gram negatives. In this study, we have investigated the molecular basis of colistin resistance in 13 MDR K. pneumoniae strains isolated from 12 patients in a teaching hospital in Sousse, Tunisia. Whole-genome sequencing (WGS) was used to decipher the molecular mechanism of colistin resistance and to identify the resistome of these CoR Kp isolates. It revealed a genome of ca. 5.5 Mbp in size with a G+C content of 57%, corresponding to that commonly observed for K. pneumoniae These isolates belonged to the 5 different sequence types (ST11, ST15, ST101, ST147, and ST392), and their resistome was composed of acquired β-lactamases, including extended-spectrum beta-lactamase and carbapenemase genes ( bla CTX-M-15 , bla OXA-204 , bla OXA-48 , and bla NDM-1 genes), aminoglycoside resistance genes [ aac ( 6 ') Ib-cr , aph ( 3 ″)- Ib , aph ( 6 )- Id , and aac ( 3 )- IIa ], and fosfomycin ( fosA ), fluoroquinolone ( qnr -like), chloramphenicol, trimethoprim, and tetracycline resistance genes. All of the isolates were identified as having a mutated mgrB gene. Mapping reads with reference sequences of the most common genes involved in colistin resistance revealed several modifications in mgrB , pmr , and pho operons (deletions, insertions, and substitutions) likely affecting the function of these proteins. It is worth noting that among the 12 patients, 10 were treated with colistin before the isolation of CoR Kp No plasmid encoding mcr-1 to mcr-5 genes was found in these isolates. This study corresponds to the first molecular characterization of a collection of CoR Kp strains in Tunisia and highlights that the small-transmembrane protein MgrB is a main mechanism for colistin resistance in K. pneumoniae ., (Copyright © 2018 American Society for Microbiology.)

This study was conducted to investigate extended-spectrum-β-lactamase (ESBL) producing Enterobacteriaceae isolates from the Center of Maternity and Neonatology of Monastir, Tunisia. Fourty-six strains out of 283 were found to produce ESBL: Klebsiella pneumoniae (n = 37), Escherichia coli (n = 6), Enterobacter cloacae (n = 2), and Citrobacter freundi (n = 1). Genotyping analysis, using ERIC2 and RAPD, showed that strains were clonally unrelated. PCR amplification followed by sequencing revealed that all strains produced CTX-M-15. This enzyme was co-produced with TEM and SHV determinants in 34 and 36 strains respectively. The bla CTXM-15 gene was bracked by ISEcp1 and/or IS26 in 42 out of the 46 ESBL positive strains. The quinolone resistance determinants were associated to the ESBL producing isolates: we identified the qnrB1 gene in six isolates and the aac(6')-Ib-cr gene in five isolates. This epidemiological study shows the widespread of CTX-M-15 and qnr determinants among enterobacterial isolates from neonates hospitalized at the center of Maternity and Neonatology of Monastir suggesting either mother portage or horizontal transmission., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Purpose: Group A rotavirus (RVA) is the leading cause of severe gastroenteritis in children younger than 5 years. The most common human G-types are G1-4 and G9. G12 genotype is currently emerging worldwide, becoming the sixth most prevalent RVA G-genotype. In Tunisia, an emergence of G12 RVA strains was observed. To understand the evolution and origin of these Tunisian G12 strains, phylogenetic analyses were conducted., Methodology: A total of 1127 faecal samples were collected from Tunisian children under 5 years consulting for gastroenteritis between 2009 and 2014. Samples were screened by ELISA for the presence of RVA antigen. RVA-positive samples were used for the detection of G12 RVA strains by semi-nested RT-PCR. G12-positive specimens were subjected to VP4 genotyping reaction. PCR products of the G12-positive samples were sequenced and characterized by phylogenetic analysis of partial VP7 gene sequence., Results: Globally, 270 (24 %) stool specimens were RVA-positive. Fourteen presented the G12 genotype (5.2 %) and were found to be in combination with either the P[6] (50.0 %) or the P[8] (50.0 %) genotype. Phylogenetic analysis revealed that all characterized Tunisian G12 strains clustered in the modern G12 lineage III and appear to form three different subclusters., Conclusion: Thus, the Tunisian G12 strains may have originated from not a single, but at least three distinct ancestral G12 strains. Detailed molecular characterization of the entire genome of these strains remains essential to help determine the extent of genetic variation and the relatedness of Tunisian G12 RVA strains to G12 strains described worldwide.

Group A rotavirus (RVA) represents the most important aetiological agent of diarrhoea in children worldwide. From January 2009 to December 2014, a multi-centre study realized through 11 Tunisian cities was undertaken among children aged <5 years consulting or hospitalized for acute gastroenteritis. A total of 1127 faecal samples were collected. All samples were screened by ELISA for the presence of RVA antigen. RVA-positive samples were further analyzed by PAGE and used for G/P-genotyping by semi-nested multiplex RT-PCR. Globally, 270 specimens (24 %) were RVA-positive, with peaks observed annually between November and March. Nine different electropherotypes could be visualized by PAGE, six with a long profile (173 cases) and two with a short one (seven cases). Mixed profiles were detected in two cases. Among the 267 VP7 genotyped strains, the predominant G- genotype was G1 (39.6 %) followed by G3 (22.2 %), G4 (13 %), G9 (11.5 %), G2 (5.2 %) and G12 (5.2 %). Among the 260 VP4 genotyped strains, P[8] genotype was the predominant (74.5 %) followed by P[6] (10.4 %) and P[4] (5.5 %). A total of 257 strains (95.2 %) could be successfully G- and P-genotyped. G1P[8] was the most prevalent combination (34.4 %), followed by G3P[8] (16.3 %), G9P[8] (10.3 %), G4P[8] (8.9 %), G2P[4] (4 %), G12P[6] (2.6 %) and G12P[8] (1.9 %). Uncommon G/Pgenotype combinations, mixed infections and untypeable strains were also detected. This is the first report, in Tunisia, of multiple detection of an emerging human RVA strain, G12 genotype. This study highlighted the need for maintaining active surveillance of emerging strains in Northern Africa.

Unlabelled: The evolution of multidrug resistance remains an alarming topic due to selection pressure related to the inappropriate use of antibiotics., Objectives: Our work is in this perspective and focuses on the evolution of the consumption of antibiotics active on gram-negative bacilli, and the evolution of bacterial resistance., Materials and Methods: International indicator of antibiotic consumption was based on the method of Defined Daily Dose reported to the number of days of hospitalization. The search for a correlation between bacterial resistance and antibiotic consumption was conducted by the Spearman test., Results: A statistically significant correlation was identified between the rates of enterobacteriaceae resistant to 3(rd) generation cephalosporin, particularly those secreting beta-lactamases with extended spectrum, and consumption of 3(rd) generation cephalosporin (p= 0.002) and imipenem (p= 0.04). Also, a statistically significant relationship between the multi-resistant bacteria and the rate of consumption of colistin (p= 0.041) and fluoroquinolones (p= 0.002) was also reported in this study., Conclusion: Monitoring of both evolution of multidrug resistance and the use of antibiotics helps us to better understand the situation and establish more efficient antibiotic protocols.

This study was conducted to identify the molecular mechanisms of imipenem resistance in a Klebsiella pneumoniae (Kp16137) isolate recovered in August 2008 at the University Hospital Sahloul, Sousse, Tunisia. The strain was identified with the API 20E system; antibiotic-containing disks were used for detection of antibiotic susceptibility by a disk diffusion assay. We investigated the presence of β-lactamases by PCR, using specific primers for bla(TEM), bla(SHV), bla(CTX-M), bla(OXA), bla(CMY), bla(ACC), bla(FOX), bla(IMP), bla(KPC), bla(VIM), and by sequencing. Extraction of plasmid DNA from Kp16137 and the transconjugant was performed by the method of Kado. Southern transfer was performed on nylon. The membrane was hybridized with a specific probe for the bla(CMY-2) gene. Outer membrane proteins were isolated and were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 12% polyacrylamide gel. K. pneumoniae Kp16137 was resistant to all available β-lactams, including third generation cephalosporins and carbapenems. The screening of β-lactamases showed the presence of three β-lactamases: TEM-1, SHV-61, and CMY-4. The CMY-4 β-lactamase was located on an 80-kb plasmid. An analysis of the outer membrane proteins of this isolate revealed that it lacked a porin of 42 kDa. The loss of this outer membrane protein band correlated with imipenem resistance in this strain. In K. pneumoniae 16137, synthesis of a plasmid-mediated β-lactamase: AmpC CMY-4, together with alteration in permeability led to resistance to all available β-lactams and carbapenems.

One hundred extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae were recovered from the intensive care unit and the urology ward of the University Hospital of Sahloul in Tunisia between May 2005 and May 2006. The majority of strains showed a high level of resistance to cefotaxime and ceftazidime. Double-disk synergy test and E-test strips were used to confirm production of ESBLs. The molecular analysis revealed that the majority of strains (91%) carried genes encoding CTX-M-15. SHV-12 and SHV-2a were produced, respectively, by 9% and 3% of the strains. Pulsed-field gel electrophoresis of ESBL-producing Klebsiella pneumoniae isolates revealed four different clonal groups and three for Escherichia coli, showing the absence of spread of any epidemic clone. The CTX-M-15 ESBL-producing E. coli of the major clonal group belong to the B2 phylogenetic group, to the sequence type 131, and has a high virulence potential. In conclusion, CTX-M-15 ESBLs accounted for the overwhelming majority of ESBL types among Enterobacteriaceae from our hospital. This study confirms the high rate of ESBLs in Tunisia and further demonstrates the worldwide spread of genes coding for CTX-M-15 enzymes in clinical isolates.

The aim of this study was to describe the distribution of the trimethoprim-sulfamethoxazole resistance genes and their association with class 1 integrons in a collection of clinical isolates of Enterobacteriaceae recovered at the University Hospital Sahloul in Tunisia. A total of 80 isolates of Enterobacteriaceae were studied, including six different species. There were 35 extended-spectrum beta-lactamases (ESBL)-producing isolates. Resistance to trimethoprim-sulfamethoxazole was assessed by the disk diffusion method. Polymerase chain reaction (PCR) with primers specific for sul1, sul2, and sul3 was used to detect the three known sulphonamide resistance genes. The presence of class 1 integrons in the studied isolates was detected using PCR and the resistance gene cassettes were characterized by directly sequencing the PCR products obtained with 5'conserved segment (5'CS) and 3'conserved segment (3'CS) primers. The int1 gene was found in 68 out of 80 enterobacterial isolates. The sul1 gene was found in 22 isolates (27.5%), sul2 gene in 5 isolates (6.25%), and both genes in 49 isolates (61.25%). Eight of the studied isolates had no dfr alleles, and in the remaining 72 isolates, 7 dfr genes were identified. The most prevalent were dfrA7 (40%) and dfrA17 (33%). Class 1 integrons were found to be an important genetic element of resistance to trimethoprim-sulfamethoxazole among the clinical isolates of Enterobacteriaceae. The types, combinations, and frequency of the gene cassettes in integrons provide useful data for the surveillance of antimicrobial resistance in our hospital and for the prescription practice of cotrimoxazole.

Extended-spectrum beta-lactamases (ESBLs) in Pseudomonas aeruginosa are increasingly reported worldwide. In our study, a total of 70 clinical isolates of multidrug-resistant P. aeruginosa were studied. Isoelectric focusing electrophoresis, PCR, and PCR product sequencing were designed to characterize the contained ESBLs. The Double Disk Synergy Test in Cloxacillin (250 microg/ml)-containing Mueller-Hinton agar plates with a 20 mm distance between disks was the most reliable ESBL-screening method. Seven out of 70 multidrug-resistant P. aeruginosa clinical isolates were positive for ESBL and have the bla(SHV-2a) ESBL gene. The bla(SHV-2a)-positive isolates were clonally related according to Enterobacterial Repetetive Intergenic Consensus-PCR (ERIC-PCR) results. The bla(SHV-2a) gene was found to be chromosomally located, and the flanking IS26 sequence in the immediate upstream region of the bla(SHV-2a) gene was detected in all SHV-2a-producing isolates. This is the first report of SHV-2a-producing P. aeruginosa isolates from Tunisia.

Among human rotaviruses, G9 has emerged as the fifth most important genotype circulating globally. Ongoing surveillance of rotavirus in Tunisia during the past 10 years identified the first G9 strains in 2004. These strains exhibited the P[8] VP4 genotype and had a long RNA electrophoretype. The G9 strains were characterized by phylogenetic analysis of the VP7 gene sequence and showed high identity with other human rotavirus G9 strains belonging to the rotavirus VP7 lineage group III.

This study was conducted to identify metallo-beta-lactamase (MBL) producers among a collection of 75 nonrepetitive carbapenem-resistant Pseudomonas aeruginosa isolates recovered between November 2003 and May 2007 at the University Hospital Sahloul, Sousse, Tunisia. Five isolates produced the MBL VIM-2. Those bla(VIM-2)-positive isolates were clonally related according to pulsed-field gel electrophoresis analysis. This carbapenemase was very likely chromosomally located and as a form of a gene cassette in a class 1 integron. This is the 1st report of spread of VIM-2 producers in Tunisia.

Ninety-nine carbapenem-resistant Acinetobacter baumannii isolates were obtained from patients hospitalized between October 2005 and January 2007 at the University Hospital Sahloul, Sousse, Tunisia. Thirteen of those isolates produced the carbapenem-hydrolyzing oxacillinase OXA-23. All the OXA-23-positive isolates were clonally related, and the bla(OXA-23) gene was found to be chromosomally located and associated with an upstream-located insertion sequence ISAba1. This study further highlights the worldwide emergence of OXA-23-producing A. baumannii isolates.

The relationships between host factors, virus strain, viral load, and illness severity in respiratory syncytial virus (RSV)-induced bronchiolitis are poorly defined. These relationships were evaluated prospectively in 81 previously healthy infants hospitalized with RSV bronchiolitis. Disease severity was determined by the respiratory rate, the duration of hospitalization, and whether patients during their hospitalization required pediatric intensive care unit admission or mechanical ventilation. RSV typing into subgroup A and B was obtained by RT-PCR-hybridization assay. The nasopharyngeal RSV viral loads were measured by real-time quantitative RT-PCR. Disease severity correlated significantly with the presence of risk factor (estimated gestational age < 37 weeks and/or birth weight < 2,500 g) and with chronologic age

Objective: to characterize epidemiological and clinical features related to the multi-drug Acinetobacter baumannii infections in the university hospital Sahloul in Tunisia., Material and Methods: retrospective study including twenty-four imipenem resistant Acinetobacter baumannii isolated from twenty patients hospitalized in different wards of the hospital. Study of clinical features related to the infection by multi-drug Acinetobacter baumannii, bacterial identification by classical identification scheme, antibiotic susceptibilities were determined by the disk diffusion method; genotyping was performed by arbitrarily-primed PCR., Results: the most incriminated ward was the intensive care unit with a high prevalence of septicaemia. All studied strains were multi-drug to all beta-lactams tested. Genotyping has shown the clonality of studied strains. Features incriminated in the acquisition of infection were essentially immunodeficiency, invasive manoeuvring and antibiotherapy., Conclusion: multidrug Acinetobacter baumannii is increasingly isolated in our hospital. Rational use of antibiotics and rigorous application of hygienic rules could contribute to limit dissemination of such strains.

Viral diarrhoea remains a major cause of childhood morbidity and mortality worldwide. Four major categories of viruses are now recognized as clinically important, including rotavirus, astrovirus, adenovirus, and calicivirus. This retrospective epidemiological study was conducted in the East centre part of Tunisia. A total of 638 stool samples were collected from children under 5 years of age presenting with acute diarrhoea at hospitals the East centre part of Tunisia between October 2003 and September 2005. All samples were analyzed using commercially available immunoenzymatic assay (EIA) kits to detect specific adenovirus antigens. Samples positive for adenovirus antigen were further screened using an ELISA technique allowing specific detection of species F enteric adenovirus types 40 and 41. Adenovirus was detected in 6% of the stools tested using ELISA. Among stool samples testing positive for adenovirus, 57% (20/35) were found to contain species F adenovirus types 40/41. In addition to diarrhoea that was present in all children studied, vomiting and fever were observed in 89% and 53% respectively and were associated with respiratory troubles in 32%. Enteric adenoviruses appear to play an important role in paediatric diarrhoea in Tunisia. Use of simple effective viral diagnostic techniques in paediatric hospitals could improve patient care by reducing unnecessary use of antibiotics.