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Aims In Tunisia, limited research has focused on characterizing clinical vancomycin-resistant Enterococcus faecium (VREfm). This study aimed to bridge this knowledge gap by molecular characterization of antimicrobial resistance, determining the genetic elements mediating vancomycin-resistance, and whole-genome sequencing of one representative VREfm isolate Methods and results Over 6 years (2011–2016), a total of eighty VREfm isolates responsible for infection or colonization were identified from hospitalized patients, with the incidence rate increasing from 2% in 2011 to 27% in 2016. All of these strains harbored the vanA gene. The screening for antimicrobial resistance genes revealed the predominance of ermB, tetM , and aac(6′)-Ie-aph(2′')-Ia genes and 81.2% of strains harbored the Tn 1545. Pulsed-field gel electrophoresis identified seven clusters, with two major clusters (belonging to ST117 and ST80) persisting throughout the study period. Seven Tn 1546 types were detected, with type VI (truncated transposon) being the most prevalent (57.5%). Whole-genome sequencing revealed a 3 028 373 bp chromosome and five plasmids. Mobile genetic elements and a type I CRISPR-cas locus were identified. Notably, the vanA gene was carried by the classic Tn 1546 transposon with ISL3 insertion on a rep17pRUM plasmid. Conclusion A concerning trend in the prevalence of VREfm essentially attributed to CC17 persistence and to horizontal transfer of multiple genetic variants of truncated vanA -Tn 1546. [ABSTRACT FROM AUTHOR]

Aims Carbapenem-resistant Acinetobacter baumannii (CR- Ab) is an important cause of infections in burn patients. This study aimed to characterize the antimicrobial susceptibility pattern of CR- Ab isolated from burns in Burn Intensive Care Unit (BICU) of the Trauma and Burn Centre of Ben Arous, to determine the prevalence of β -lactamase-encoding genes and to search eventual genetic relatedness of CR- Ab strains. Methods and Results From 15 December 2016 to 2 April 2017, all nonduplicated CR- Ab isolated in burn patients in the BICU were screened by simplex Polymerase Chain Reaction (PCR) for the class A, B, C, and D β -lactamase genes. Sequencing was performed for NDM gene only. Genetic relatedness was determined by using pulsed field gel electrophoresis (PFGE) and by multilocus sequence typing. During the study period, 34 strains of CR- Ab were isolated in burns, mainly in blood culture (n  = 14) and central vascular catheter (n  = 10). CR- Ab strains were susceptible to colistin but resistant to amikacin (91%), ciprofloxacin (100%), rifampicin (97%), and trimethoprim-sulfamethoxazole (100%). All strains harbored bla OXA-51-like and bla OXA-23 genes, only or associated to bla GES (n  = 26; 76%), bla ADC (n  = 20; 59%), bla PER-1 (n  = 6; 18%) or/and bla NDM-1 (n  = 3; 9%). PFGE identified 16 different clusters and revealed that most strains belonged to one major cluster A (n  = 15; 44.1%). Among NDM-1 isolates, two were clonally related in PFGE and belonged to two single locus variant sequence type ST-6 and ST-85. Conclusions This is the first description of clonally related NDM-1 and OXA-23-producing A. baumannii strains in the largest Tunisian BICU associated with two single locus variant sequence types ST6 and ST85. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii has emerged as a main nosocomial pathogen exhibiting high rates of resistance to clinically relevant antibiotics. Six pandrug-resistant A. baumannii (PDR-A. baumannii) were recovered from three patients in a Tunisian Intensive Care Unit (ICU) between 10th and 16th of May 2018 resulting in one fatal case and raising the possibility of an outbreak. On 18th of May environmental screening of ICU surfaces was carried out. On 22nd of May a fourth patient was infected with PDR-A. baumannii and died. A second investigation was carried out for environmental screening and PDR-A. baumannii was isolated from the respirator. Antimicrobial susceptibility testing was performed according to EUCAST (2019) guidelines. MIC of colistin was determined by broth microdilution method. PCR was used to detect 14 beta-lactamases/carbapenemases and mcr (mcr-1 to mcr-5) genes. The genetic relatedness of PDR-A. baumannii isolates was determined by PFGE and MLST. Seven PDR-A. baumannii isolates were recovered from four patients, one MDR strain from wash basin, a PDR strain from hand sanitizer bottle and another PDR strain from respirator. All PDR-A. baumannii (n 5 9) harbored blaOXA-69 gene and none carried mcr. Moreover, seven carried blaGES and blaOXA-23 genes. PFGE identified four pulsotypes (A, B, C, and D) with the pulsotype A gathering seven PDR-A. baumannii isolates: six from three patients and one from hygiene sample. MLST revealed that all PDR-A. baumannii isolates of pulsotype A belonged to the pandemic clone ST2. Systematic screening of MDR and PDR-A. baumannii is highly recommended to limit dissemination of such strains in ICUs. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is one of the major infectious agents in burn patients. Globally, high rates of antimicrobial resistance in P. aeruginosa have been reported, which is a cause of concern. The objective of this study was to determine the rate of resistance to carbapenems in P. aeruginosa isolates recovered from burn patients in Tunisia, to search genes encoding for carbapenemases and to determine their epidemiological markers (serotypes). A retrospective study was conducted in the Burn Intensive Care Unit (BICU) of the Trauma and Burn Centre of Ben Arous, Tunisia, and P. aeruginosa isolates collected from burn patients, from January to December 2018 were investigated. Carbapenemase screening was performed by Carbapenem Inactivation Method (CIM) and by EDTA-disk test for all carbapenem resistant isolates. Genes encoding carbapenemases (bla VIM, bla IMP, bla GES, bla NDM, and bla KPC) were investigated by PCR and selected carbapenemase genes were sequenced. During the study period, 104 non duplicated P. aeruginosa isolates were recovered. Most of them were isolated from skin samples (45.1%) and blood culture (22.1%) and belonged to O:11 (19.2%), O:12, and O:5 (12.5%, each) serotypes. High rates of resistance were observed for carbapenems (64.4%). Among the 67 carbapenem resistant isolates, 58 (86.5%) harbored bla VIM gene and 55 (82%) bla GES gene; in addition, 48 (71.6%) co-harbored bla VIM and bla GES genes. After sequencing, the bla VIM-2 and bla GES-5 gene variants were identified in seven randomly selected isolates. To the best of our knowledge, this is the first description of P. aeruginosa simultaneously harboring bla VIM-2 and bla GES-5 genes. [ABSTRACT FROM AUTHOR]

Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen causing health care-infections in the world, especially in burns. The aim of this study was to assess the extent of dissemination of MRSA isolated from burn patients in Burn Intensive Care Unit in Tunisia and to evaluate the frequency of virulence and antibiotics resistance genes. Among the 72 S. aureus isolates analyzed in the study, 54% were MRSA. The majority of MRSA (94.8%) were multidrug resistant and they had a high resistance rates to kanamycin (94.8%), tobramycin (90%), tetracycline (94.8%) and ciprofloxacin and rifampicin (87%, each). The gene aac(6′)-Ie-aph(2″)-Ia conferring resistance to kanamycine and tobtamycin were detected in all isolates and the aph(3′)-Ia gene conferring resistance to gentamicin were detected in 2.8% of resistant isolates. Tetracycline resistance genes tet(M), tet(K) and tet(L) were detected in 100%, 10.8% and 2.8% of the isolates, respectively. The SCC mec type III and the agr type I were the most predominant (69.2% and 90%, respectively). The 27 SCC mec III- agr I isolates were clustered into two PFGE types A and B. The two representative isolates of PFGE clusters A and B belonged to ST239-t037 and ST241-t037 respectively. As conclusion, our results showed a high prevalence of MRSA in trauma burn intensive care unit belonging to two multidrug resistant clones ST239/ST241- agr I-t037-SCC mec III MRSA. We also demonstrated that MRSA was disseminated between burn patients. [ABSTRACT FROM AUTHOR]

This study aimed to characterize the first vancomycin-resistant Enterococcus faecalis (VREfs) isolate from patient with neutropenic in Tunisia by whole-genome sequencing (WGS). This strain was detected from routine rectal swab from an 8-year-old child with bone marrow aplasia, residing in a rural area, on September 20, 2021. The strain was isolated after 12 days of hospitalization at the National Bone Marrow Transplant Center. Minimum Inhibitory Concentrations of vancomycin and teicoplanin were >256 and 16 mg/L, respectively. WGS revealed that the strain belonged to the ST249 clone, exclusively reported in avian (poultry and ducks) vancomycin-susceptible E. faecalis isolates in six studies from four countries, primarily Denmark. The vanA gene was carried by the Tn1546 transposon mobilized by a pTW9-like plasmid. The ardA gene, a CRISPR-Cas system neutralization factor, was detected in this strain. In summary, this is the first report of avian-associated E. faecalis ST249 in clinical samples. Initially vancomycin susceptible, the strain acquired a pTW9-like plasmid carrying the classical vanA-Tn1546 transposon. This acquisition was facilitated by the sex pheromone-response mechanisms and the ardA gene and CRISPR-Cas system neutralization. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a common pathogen causing hospital infections. The increasing rate of healthcare-associated infections caused by methicillin-resistant Staphylococcus aureus (MRSA) in developing countries has led to many public health problems. This study aimed to investigate the molecular epidemiology as well as the antibiotic resistance pattern of clinical isolates of MRSA from Southern Iran. A total of 135 S. aureus isolates were collected from the patients referred to three hospitals in South Iran. The phenotypic and genotypic diagnosis of MRSA isolates was performed by disk diffusion and PCR methods, respectively. The antibiotic resistance pattern for MRSA isolates was performed using Kirby–Bauer method. The molecular epidemiology of isolates was performed by MLST, Spa typing and SCC mec typing. From 135 S. aureus isolates, 50 (37%) MRSA strains were detected from which two different sequence types including ST239 and ST605 were identified. SCC mec type III was the most common profile (50%) and t030 was the predominant spa type (48%) among the strains. The MRSA isolates had the highest resistance to penicillin (100%), tetracycline (88%), levofloxacin (86%), ciprofloxacin (84%), erythromycin (82%), gentamicin (80%), and clindamycin (78%). The results of this study show that the most common genetic type among the MRSA isolates was ST239-SCC mec III/t030. The rapid and timely detection of MRSA and the administration of appropriate antibiotics according to the published antibiotic resistance patterns are essential. Furthermore, the continuous and nationwide MRSA surveillance studies are necessary to investigate clonal distribution and spreading of MRSA from community to hospitals. [ABSTRACT FROM AUTHOR]

Cytomegalovirus reactivation (CMV-R) is a frequent complication post-allogeneic hematopoietic stem cell transplantation (allo-HSCT), associated with poor outcomes. Previous studies have demonstrated the protective effect of CMV-R against relapse after allo-HSCT for acute myeloblastic leukemia (AML). However, this impact remains unclear in acute lymphoblastic leukemia (ALL). We conducted a retrospective study on 81 patients with ALL who received allo-HSCT after myeloablative conditioning regimen from matched sibling donors between 2016 and 2022. All patients underwent weekly monitoring for CMV-R by quantitative polymerase chain reaction assay from engraftment until day +100 post allo-HSCT, and received antiviral prophylaxis with acyclovir from day +1 to 6 months after allo-HSCT. Preemptive treatment was initiated when a viremia was higher than 150 copies/mL. The median age was 20 years (range, 5-50 years). After allo-HSCT, 35% of patients developed CMV-R after a median of 39 days (range, 19-100 days). After a median follow-up of 30 months (range, 1-93 months), CMV-R was an independent factor associated with lower cumulative incidence of relapse (CIR) (OR: 0.17; 95% CI [0.03 - 0.98], p = 0.04) without survival benefit. Further studies are needed to validate the protective effect of CMV-R on ALL relapse., Competing Interests: The authors declare no competing financial interests

Acinetobacter baumannii has emerged as a main nosocomial pathogen exhibiting high rates of resistance to clinically relevant antibiotics. Six pandrug-resistant A. baumannii (PDR-A. baumannii) were recovered from three patients in a Tunisian Intensive Care Unit (ICU) between 10th and 16th of May 2018 resulting in one fatal case and raising the possibility of an outbreak. On 18th of May environmental screening of ICU surfaces was carried out. On 22nd of May a fourth patient was infected with PDR-A. baumannii and died. A second investigation was carried out for environmental screening and PDR-A. baumannii was isolated from the respirator. Antimicrobial susceptibility testing was performed according to EUCAST (2019) guidelines. MIC of colistin was determined by broth microdilution method. PCR was used to detect 14 beta-lactamases/carbapenemases and mcr (mcr-1 to mcr-5) genes. The genetic relatedness of PDR-A. baumannii isolates was determined by PFGE and MLST. Seven PDR-A. baumannii isolates were recovered from four patients, one MDR strain from wash basin, a PDR strain from hand sanitizer bottle and another PDR strain from respirator. All PDR-A. baumannii (n = 9) harbored blaOXA-69 gene and none carried mcr. Moreover, seven carried blaGES and blaOXA-23 genes. PFGE identified four pulsotypes (A, B, C, and D) with the pulsotype A gathering seven PDR-A. baumannii isolates: six from three patients and one from hygiene sample. MLST revealed that all PDR-A. baumannii isolates of pulsotype A belonged to the pandemic clone ST2. Systematic screening of MDR and PDR-A. baumannii is highly recommended to limit dissemination of such strains in ICUs.