Evaluation of the implementation of the objective structured clinical examination in health sciences education from a low-income context in Tunisia: A cross-sectional study.

Background: Objective structured clinical examination (OSCE) is well-established and designed to evaluate students' clinical competence and practical skills in a standardized and objective manner. While OSCEs are widespread in higher-income countries, their implementation in low-resource settings presents unique challenges that warrant further investigation. Aim: This study aims to evaluate the perception of the health sciences students and their educators regarding deploying OSCEs within the School of Health Sciences and Techniques of Sousse (SHSTS) in Tunisia and their efficacity in healthcare education compared to traditional practical examination methods. Methods: This cross-sectional study was conducted in June 2022, focusing on final-year Health Sciences students at the SHSTS in Tunisia. The study participants were students and their educators involved in the OSCEs from June 6th to June 11th, 2022. Anonymous paper-based 5-point Likert scale satisfaction surveys were distributed to the students and their educators, with a separate set of questions for each. Spearman, Mann-Whitney U and Krusakll-Wallis tests were utilized to test the differences in satisfaction with the OSCEs among the students and educators. The Wilcoxon Rank test was utilized to examine the differences in students' assessment scores in the OSCEs and the traditional practical examination methods. Results: The satisfaction scores were high among health sciences educators and above average for students, with means of 3.82 ± 1.29 and 3.15 ± 0.56, respectively. The bivariate and multivariate analyzes indicated a significant difference in the satisfaction between the students' specialities. Further, a significant difference in their assessment scores distribution in the practical examinations and OSCEs was also demonstrated, with better performance in the OSCEs. Conclusion: Our study provides evidence of the relatively high level of satisfaction with the OSCEs and better performance compared to the traditional practical examinations. These findings advocate for the efficacy of OSCEs in low-income countries and the need to sustain them.

Impact of Covid 19 on Primary healthcare in Tunisia.

INTRODUCTION: Public primary healthcare facilities, the cornerstone of the Tunisian health system, were impacted by the Covid 19 crisis as all health systems in the world. AIM: This study aims to assess this impact of the pandemic. METHODS: We analyzed the budgetary evolution of the basic healthcare group (BHG) of Medenine and Djerba between 2019 and 2020. Similarly, we examined the evolution of all the activities of BHG of Medenine. This analysis was also completed by a semi-structured questioning with a regional expert. RESULTS: Our results showed an increase in expenditure, a decrease in revenue (12.4% for GSB of Medenine and 10.8% for BHG of Djerba), and an accumulation of outstanding payment. BHG's activities have been affected by the pandemic. We showed that stomatology and vaccine activities were well maintained during the pandemic. However, we noted a regression in the number of patients and some illnesses. Activity related to child health and health education has significantly decreased. This impact has had and will have repercussions on the epidemiological state of the population. Despite the intervention of the regional management in terms of organization, training and strengthening of equipment and human resources, the pandemic has generally impacted the operation process of these establishments, which are already facing several challenges. CONCLUSION: We recommend above all to activate the already existing opportunities to replenish the financial resources of primary healthcare facilities, to improve work environment and continuous professional development and to computerize the data and its analysis according to a scientific approach.

Staphylococcus aureus Ventilator-Associated Pneumonia: A Study of Bacterio-Epidemiological Profile and Virulence Factors.

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.

Elizabethkingia meningoseptica: An Uncommon Cause of Peritonitis In Continuous Ambulatory Peritoneal Dialysis: A Case Report and Review of Literature.

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.

Dynamics and molecular features of OXA-48-like-producing Klebsiella pneumoniae lineages in a Tunisian hospital.

OBJECTIVES: The aim of this study was to elucidate the molecular features of genes, plasmids and clones of OXA-48-like producingKlebsiella pneumoniae isolates recovered in Sahloul Hospital (Sousse, Tunisia) in the period 2012-2014. METHODS: In vitro antimicrobial susceptibility testing, S1 nuclease pulsed-field gel electrophoresis (S1-PFGE), Southern blotting and PCR-based replicon typing (PBRT) were performed. Extended-spectrum ß-lactamase (ESBL) and carbapenemases genes were detected by PCR and sequencing. The clonality of isolates was assessed by PFGE and multilocus sequence typing (MLST). RESULTS: Klebsiella pneumoniae accounted for 26.8% (1095/4083) of clinical Enterobacterales isolates identified during 2012-2014, of which 21.9% (240/1095) were resistant to carbapenems, mostly harbouring blaOXA-48-like genes (196/240; 81.7%). Plasmid analysis showed that blaOXA-204 and blaOXA-48 were mostly carried by IncA/C and IncL plasmids, respectively. The current data highlight the dominance of two ST101 and ST147 lineages spreading OXA-48 and OXA-204, respectively, through successive clonal spreads at this hospital. In addition, a large diversity of other K. pneumoniae lineages was also identified, such as ST15, ST36 and ST525 spreading OXA-48 as well as ST340, ST2032, ST301, ST199 and ST1561 spreading OXA-48 or OXA-204, constituting a reservoir of possible dominant clones in the future. CONCLUSION: This study reports the full molecular characterisation of carbapenem resistance in K. pneumoniae and the predominance of a few clones responsible for the dissemination of OXA-48 and OXA-204 enzymes in a Tunisian hospital.

Epidemiology of resistance and phenotypic characterization of carbapenem resistance mechanisms in Klebsiella pneumoniae isolates at Sahloul University Hospital-Sousse, Tunisia.

OBJECTIVE: To assess the prevalence of ESBL producing and carbapenem resistant Klebsiella pneumoniae isolated from in-come and out-come patients at Sahloul-university hospital. METHODS: A retrospective study over a 3 years period (January 2012 and December 2014) focused on 2160 strains of Klebsiella pneumoniae. Statistical analysis was carried out using SPSS program. ESBL detection was performed using a double disc diffusion method and carbapenemase detection was realized by Rosco-Disk kit. RESULTS: A total of 2160 Klebsiella pneumoniae strains were isolated during the period of the study, 26.2% (n=566) were ESBL-producers and 15.8% (n=342) showed resistance to carbapenem. The wards most affected by these strains were basically urology and intensive care units. Eighty four percent of studied strains (203/241) were resistant to temocillin, which correlate with the production of a class D (OXA-48-like) carbapenemase and 7% (17/241) showed sensitivity to EDTA and dipicolinic acid, which indicate the production of metallo-enzyme. The rate of resistance to colistin remains low. CONCLUSION: Resistance of Enterobacteriaceae, including K. pneumoniae, to third generation cephalosporins (3rd GC) and carbapenem through the mechanism of ESBL and carbapenemases production is becoming increasingly worrying. This suggests a more rational use of antibiotics, as well as the rigorous application of hygiene measurement.

Epidemiology and Whole-Genome Analysis of NDM-1-Producing Klebsiella pneumoniae KP3771 from Tunisia.

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 blaNDM-1 and blaCTX-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.

Spread of blaCTX-M-15-Producing Enterobacteriaceae and OXA-23-Producing Acinetobacter baumannii Sequence Type 2 in Tunisian Seafood.

Bivalves are filter-feeding animals and markers of bacterial pollution. We report a massive spread of blaCTX-M-15 through dominant Escherichia coli and Klebsiella pneumoniae lineages and/or plasmid subtypes (F31:A4:B1) as well as the presence of OXA-23-producing Acinetobacter baumannii sequence type 2 (ST2) in seafood, highlighting a direct risk for the consumer. These findings should urge authorities to consider hospital effluents, and also farm and urban effluents, as important sources of extended-spectrum-beta-lactamase (ESBL)/carbapenemase producers that filter-feeding animals can concentrate and further spread to humans.

Whole-genome sequencing of NDM-1-producing ST85 Acinetobacter baumannii isolates from Tunisia.

BACKGROUND: New Delhi metallo-ß-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 blaNDM-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 1st June 2013 and 31st 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 blaNDM-1, four of 246 positive for blaNDM-1 and blaOXA-23, 231 positive for blaOXA-23). Conjugation and electroporation experiments suggested that the blaNDM-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 blaNDM-1 gene revealed that the gene was located within a truncated isoform of Tn125 transposon (ΔTn125). The blaOXA-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 blaNDM-1 gene in A. baumannii was linked to clonal spread in this study.

Genomic analysis of in vivo acquired resistance to colistin and rifampicin in Acinetobacter baumannii.

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 blaOXA-23-like carbapenemase gene. Kieser extraction revealed an ca. 74 kb plasmid harbouring blaOXA-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 ß-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.

Genomic Insights into Colistin-Resistant Klebsiella pneumoniae from a Tunisian Teaching Hospital.

The emergence of colistin-resistant Klebsiella pneumoniae (CoRKp) 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 CoRKp 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 ß-lactamases, including extended-spectrum beta-lactamase and carbapenemase genes (blaCTX-M-15, blaOXA-204, blaOXA-48, and blaNDM-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 CoRKp 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 CoRKp strains in Tunisia and highlights that the small-transmembrane protein MgrB is a main mechanism for colistin resistance in K. pneumoniae.

Outbreak of colistin-resistant carbapenemase-producing Klebsiella pneumoniae in Tunisia.

OBJECTIVES: Mechanisms of colistin and carbapenem resistance among a collection of Klebsiella pneumoniae isolates recovered in a university hospital in Tunisia were studied. METHODS: In vitro antimicrobial susceptibility testing, S1 nuclease pulsed-field gel electrophoresis (S1-PFGE), Southern blotting and PCR-based replicon typing (PBRT) were performed. Extended-spectrum ß-lactamases (ESBLs), carbapenemases, AmpC-type enzymes and mgrB genes were detected by PCR and sequencing. Clonality of isolates was assessed by PFGE and multilocus sequence typing (MLST). RESULTS: Of 940 Enterobacteriaceae isolates recovered from June 2015 to March 2016 in Tahar Sfar Hospital (Mahdia, Tunisia), 220 were identified as K. pneumoniae, among which 29 were carbapenem-resistant. Carbapenem resistance was mostly due to expression of blaOXA-48 or blaOXA-204 in combination with blaCMY-4. Seven isolates carried blaNDM-1, of which two also harboured blaOXA-48, together with blaCMY-16 in one of them. All but two isolates also harboured blaCTX-M-15. All 20 blaOXA-48 genes were part of transposon Tn1999 on an IncL plasmid, whereas blaOXA-204 was found on transposon Tn2016 on an IncA/C plasmid. Finally, all blaNDM-1 genes were located within a Tn125 transposon on an IncFIIk plasmid. Interestingly, 7 (24.1%) of 29 carbapenem-resistant isolates were resistant to colistin, of which 6 were assigned to ST101, had similar PFGE profiles and presented the same 2-kb insertion in the mgrB gene. CONCLUSIONS: This study reports, for the first time in Tunisia, the full molecular characterisation of colistin resistance in K. pneumoniae. There is an urgent need for control measures and prudent use of colistin in treatment of infections with carbapenemase-producing K. pneumoniae.

Extended-spectrum ß-lactamases and plasmid-mediated quinolone resistance in enterobacterial clinical isolates from neonates in Tunisia.

This study was conducted to investigate extended-spectrum-ß-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 blaCTXM-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.

Nosocomial urinary tract infections caused by extended-spectrum beta-lactamase uropathogens: Prevalence, pathogens, risk factors, and strategies for infection control.

INTRODUCTION: Our goal was to investigate the prevalence and antibiogram pattern of extended spectrum beta-lactamase (ESBL) production among uropathogens using isolates from urine samples collected at the Department of Urology in the Sahloul Hospital, Tunisia We also aimed to identify the risk factors for nosocomial urinary tract infections (UTIs) in patients who underwent transurethral resection of the prostate (TURP) and the measures for infection control. METHODS: Laboratory records of a five-year period from January 2004 to December 2008 were submitted for retrospective analysis to determine the incidence of ESBL infections. A total of 276 isolates were collected. A case-control study involving comparisons between two groups of patients who underwent TURP was performed to determine the risk factors for ESBL infection. Group 1, designated case subjects, included 51 patients with nosocomial UTI after TURP. Group 2, designated control subjects, consisted of 58 randomly selected patients who underwent TURP without nosocomial UTI in the same period. Factors suspected to be implicated in the emergence of ESBL infection were compared between the two groups in order to identify risk factors for infection. A univariate regression analysis was performed, followed by a multivariate one. RESULTS: The annual prevalence of ESBL infection ranged from 1.3-2.5%. After performing univariate and multivariate regression analysis, the main risk factors for ESBL infections were identified as: use of antibiotics the year preceding the admission, duration of catheter use, and bladder washout (p=0.012, p=0.019, and p<0.001. CONCLUSIONS: Urologists have to perform a good hemostasis, especially in endoscopic resections, in order to avoid bladder irrigation and bladder washout and to reduce the time of bladder catheterization, which is a strong risk factor of nosocomial UTIs.

Impact of food animal trade on the spread of mcr-1-mediated colistin resistance, Tunisia, July 2015.

We report a high prevalence of MCR-1 and CTX-M-1-producing Escherichia coli in three Tunisian chicken farms. Chickens were imported from France or derived from French imported chicks. The same IncHI2-type plasmid reported to carry those genes in cattle in France and in a food sample in Portugal was found in Tunisian chickens of French origin. This suggests a significant impact of food animal trade on the spread of mcr-1-mediated colistin resistance in Europe.

Imipenem resistance in Klebsiella pneumoniae is associated to the combination of plasmid-mediated CMY-4 AmpC ß-lactamase and loss of an outer membrane protein.

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 ß-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 ß-lactams, including third generation cephalosporins and carbapenems. The screening of ß-lactamases showed the presence of three ß-lactamases: TEM-1, SHV-61, and CMY-4. The CMY-4 ß-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 ß-lactamase: AmpC CMY-4, together with alteration in permeability led to resistance to all available ß-lactams and carbapenems.