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Background: Actually, no data on the prevalence of plasmid colistin resistance in Tunisia are available among clinical bacteria. Objectives: This study aimed to investigate the current epidemiology of colistin resistance and the spread of the mcr gene in clinical Gram-negative bacteria (GNB) isolated from six Tunisian university hospitals. Methods: A total of 836 GNB strains were inoculated on COL-R agar plates with selective screening agar for the isolation of GNB resistant to colistin. For the selected isolates, mcr genes, beta-lactamases associated-resistance genes and molecular characterisation were screened by PCRs and sequencing. Results: Colistin-resistance was detected in 5.02% (42/836) of the isolates and colistin-resistant isolates harboured an ESBL (blaCTX-M-15) and/or a carbapenemase (blaOXA-48, blaVIM) encoding gene in 45.2% of the cases. The mcr-1 gene was detected in four E. coli isolates (0.59%) causing urinary tract infections and all these isolates also contained the blaTEM-1 gene. The blaCTX-M-15 gene was detected in three isolates that also carried the IncY and IncFIB replicons. The genetic environment surrounding the mcr-carrying plasmid indicated the presence of pap-2 gene upstream mcr-1 resistance marker with unusual missing of ISApl1 insertion sequence. The Conclusions: This study reports the first description of the mcr-1 gene among clinical E. coli isolates in Tunisia and provides an incentive to conduct routine colistin susceptibility testing in GNB clinical isolates. [ABSTRACT FROM AUTHOR]

Background: Since 2012, few reports on the molecular epidemiology of Pseudomonas aeruginosa were reported in Tunisia. Objectives: This study aimed to evaluate carbapenem-resistance determinants and molecular epidemiology and to compare the carbapenemase-phenotypic detection methods of multidrug-resistant P. aeruginosa isolates. Methods: During a period of four years (2014 to 2017), all imipenem-ceftazidime-resistant P. aeruginosa isolates were retrospectively selected at the microbial laboratory of Charles Nicolle hospital of Tunis. These isolates were examined by the modified Hodge test, modified carbapenem inactivation method (mCIM), and another mCIM, called CIMTris, and their performance was evaluated using PCR analysis as the gold standard. Results: A total of 35 isolates were recovered among patients hospitalized in different units. All strains were colistin-susceptible.All carbapenem-resistant isolates showed a high-level resistance to carbapenems. CIMTris and mCIM showed 96.15% and 46.15% sensitivity and 44.44% and 100% specificity, respectively, for detecting carbapenemase production.Conclusions: CIMTris is a promising approach for detecting carbapenemase activity in P. aeruginosa and merits further testing. Moreover, this study described the first detection of GES-5- and GES-9-producing P. aeruginosa in Tunisia as well as the co-occurrence of the blaGES-5 and blaVIM-11 carbapenemase genes in one isolate. These findings are of great concern because the rapid dissemination of MDR strains represents a major therapeutic and epidemiological threat. [ABSTRACT FROM AUTHOR]

• Real time PCR is increasingly used for the diagnosis of invasive bacterial infections • Real time PCR was implemented in three North African countries for surveillance • Quality assurance schemes were also implemented • The results suggest the predominance of Neisseria meningitidis group B We implemented a project named MENINGSTOP in three countries of North Africa (Algeria, Morocco and Tunisia). The main objective was to use real-time PCR to detect, identify and type the three main agents (Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae) responsible for invasive bacterial infections (IBI). The protocol of WHO and US CDC for real-time PCR was used to detect and type the three bacterial agents in clinical samples. We also designated two quality exercises using an external interlaboratory study and cross-testing of 10% of randomly selected samples. Among the 752 samples tested, 18% were positive for one of the three agents. N. meningitidis was the most frequent globally reaching 9% of all samples (7% to 17% range) followed by S. pneumoniae 8% of all samples (6% to 15%). Group B meningococci was the most frequent (74% of all positive samples for meningococci and ranging from 50% to 90%). Quality assurance showed >85% correlation scores. Real-time PCR can help improving epidemiological surveillance. Data confirm the prevalence of meningococci B. Our project adds a reliable tool to enhance surveillance and to help decision making in vaccination strategies against IBI. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a feared pathogen in the burn center due to its opportunistic nature and its multidrug resistance. Our purpose was to study the incidence density of Acinetobacter baumannii colonization and infection and to determine the antibiotic susceptibility of the strains isolated in patients hospitalized in the Trauma and Burn Center Burn Unit in Tunisia. Our retrospective study included 1517 non-repetitive strains of Acinetobacter baumannii, between January 2012 and September 2020, with an average rate of 12.2% of the service's bacterial ecology. The incidence density of Acinetobacter baumannii colonization and infection was 13.7? days of in-patient stay and 14.1? days of in-patient stay, respectively. A positive and statistically significant correlation between Acinetobacter baumannii colonization and infection (rs=0,7; p=0.005) was noted in our study. The colonization strains were mainly isolated from central catheters (71.2%) and skin swab samples (22.9%). Infections were dominated by bacteremia (47.6%) and respiratory tract infections (25.4%). Bacteremia was microbiologically documented in 53% of cases. The most common source of bacteremia was central catheters (60.8%), skin (22.2%) and respiratory tract (15.5%). The rates of resistance inAcinetobacter baumannii to the antimicrobial agents tested were high: ceftazidime (85.2%), pipéracillin-tazobactam (95.6%), imipenem (95.3%), amikacine (91.1%), ciprofloxacin (93.5%), rifampicin (36.4%) and cotrimoxazole (88.1%). The resistance of colistin was noted in 1.8% of cases. [ABSTRACT FROM AUTHOR]

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Carbapenemase-producing Enterobacteriaceae (CPE) pose an emerging threat worldwide. The purpose of our work was to determine their prevalence among patients hospitalized in the Trauma and Burn Centre's Burn Unit, and to study their microbiological and molecular characteristics. Our study was longitudinal, conducted at the Trauma and Burn Centre's laboratory between January and June 2017. It focused on 42 CPE strains isolated from 34 patients. These strains were made up of K. pneumoniae (24), P. mirabilis (13), K. oxytoca (2), P. stuartii (1), E. coli (1), and E. cloacae (1). The overall prevalence of CPE among Enterobacteriaceae was 14.43% with a monthly decline. The blaNMD gene (59%) predominated on blaOXA-48 (33%) and 7% of strains co-expressed these two genes. We describe the first case of P. stuartii carrying blaNDM in Tunisia. Resistance to ertapenem, imipenem and meropenem was 83%, 57%, and 10% respectively. The antibiotics showing the highest resistant rates were third-generation cephalosporins (97%), gentamicin (98%) and norfloxacin (90%). Colistin and fosfomycin had the best-preserved activity in vitro with 4% and 33% resistance, respectively. The prevalence of EPCs is high among burns. Screening efforts, hygiene measures and the preservation of the few molecules still active are a vital issue. [ABSTRACT FROM AUTHOR]

Highlights • High prevalence of imported/acquired carbapenem-resistant A. baumannii (CRAB) faecal carriage among ICU patients. • OXA-23 was the most frequent carbapenemase among identified CRAB, followed by NDM-1. • Co-occurrence of carbapenem and aminoglycoside resistance genes among multidrug-resistant A. baumannii. • Circulation of clonally related CRAB strains. ABSTRACT Gastrointestinal colonisation by carbapenem-resistant Acinetobacter baumannii (CRAB) is a critical step before nosocomial infection. This study evaluated CRAB intestinal carriage in patients admitted to a Tunisian ICU and determined the antimicrobial resistance mechanisms involved. From December 2014 to February 2015, all 63 patients admitted to the ICU were screened for rectal CRAB colonisation upon admission and once weekly thereafter. ICU patients who acquired a CRAB nosocomial infection were also included. β-Lactamases and associated resistance genes were screened by PCR sequencing, and molecular typing was performed by PFGE and MLST. The CRAB faecal carriage rate at admission was 4.8% (3/63). The CRAB acquisition rate during ICU stay was analysed in 39 of the remaining 60 patients and the rate of acquired CRAB faecal carriage was 15.4% (6/39); 4 patients also showed an ICU-acquired CRAB infection (one patient was a faecal carrier and suffered infection). Overall, 13 CRAB isolates were collected from 12 patients, of which 11 isolates showed resistance to all antibiotics tested except colistin. bla OXA-23 and bla NDM-1 were detected in 11 and 2 isolates, respectively. All OXA-23-producing strains carried armA, tetB, sul1 and catB , and some of them carried aph(3′)-VIa, bla TEM-1 , aph(3′)-Ia and ant(2′′)-Ia. The bla NDM-1 -positive isolates harboured aph(3′)-VIa and catB. Three PFGE patterns and two STs were identified [ST195 (n = 11), ST1089 (n = 2, NDM-1-positive)]. Whether imported or acquired during ICU stay, CRAB colonisation is a major risk factor for the occurrence of serious nosocomial infection. Systematic screening of faecal carriage is mandatory to prevent their spread. [ABSTRACT FROM AUTHOR]

Our study aimed to investigate colistin resistance and the mechanisms involved in a collection of 35 extended-spectrum beta-lactamase (ESBL) and 13 CMY-2-producing E . coli strains which were previously recovered from chicken gut microbiota in Tunisia, as well as to determine the genetic location of mcr genes. Forty-eight ESBL and CMY-2-producing E . coli strains were obtained from 137 fecal samples of healthy chickens during 2013. These strains were tested for colistin resistance by the broth microdilution method, and screened for mcr-1 and mcr-2 genes by PCR. Two of these strains were colistin-resistant (MIC = 8 mg/L). Both harbored the mcr-1 gene, were CMY-2 producers, and were additionally resistant to tetracycline, ciprofloxacin, chloramphenicol, gentamicin, tobramycin and trimethoprim-sulfamethoxazole. They shared phylogroup A, the same pulsed-field gel electrophoresis (PFGE)-pattern, and were typed as ST2197. In both strains, IS Apl1 and pap2 were detected upstream and downstream of mcr-1 gene, respectively. The analysis of the two mcr-1 -positive strains and their transconjugants by PCR-based replicon typing and S1-PFGE, demonstrated that mcr-1 gene is linked to an IncP plasmid (~242 kb), and bla CMY-2 to an IncI1 plasmid (97 kb). The occurrence of E . coli harboring mcr-1 gene among intestinal microbiota in poultry and its location on a conjugative plasmid could represent a risk for public health. The evolution of this type of resistant microorganisms should be evaluated in the future. [ABSTRACT FROM AUTHOR]

This study was conducted to detect extended spectrum beta-lactamases (ESBLs) and plasmidic AmpC beta-lactamase (pAmpC-BL)-producing Escherichia coli isolates in industrial poultry samples were collected from healthy chickens of the three farms. Samples were inoculated onto desoxycholate-lactose-agar plates supplemented with cefotaxime (2 mg/L). E. coli was identified by biochemical and molecular methods and antibiotic susceptibility testing by the disk diffusion method. Genes encoding ESBLs and pAmpC-BL were detected by PCR and sequencing. Phylogenetic groups were determined by triplex PCR. The molecular typing of strains was done by pulsed field gel electrophoresis (PFGE) and Multilocus Sequence Typing (MLST) in those isolates showing different PFGE patterns. Cefotaxime-resistant E. coli isolates were recovered in 48 of 137 fecal samples (35%), and one isolate/sample was further studied. The following beta-lactamase genes were detected: bla CTX-M-1 (29 isolates, isolated in all three farms), bla CTX-M-15 (5 isolates, confined in farm II), bla CTX-M-14 and bla CMY-2 (one isolate and 13 isolates, respectively, in farm III). The 48 cefotaxime-resistant isolates were distributed into phylogroups: B1 (n = 21), A (n = 15) and D (n = 12). PFGE analysis revealed 19 unrelated patterns: 15 different profiles among ESBL-positive strains and 4 among the CMY-2-positive isolates. The following sequence types-associated phylogroups were detected: a) CTX-M-1-positive strains: lineages ST542-B1, ST212-B1, ST58-B1, ST155-B1 and ST349-D; b) CTX-M-15-positive strain: lineage ST405-D; c) CTX-M-14-positive strain: lineage ST1056-B1; d) CMY-2-positive strains: lineages ST117-D, ST2197-A, and ST155-B1. Healthy chickens constitute an important reservoir of ESBL- and pAmpC-BL-producing E. coli isolates that potentially could be transmitted to humans via the food chain or by direct contact. [ABSTRACT FROM AUTHOR]

This paper investigates the demand function for residential water in Tunisia, using the discrete/continuous choice model. The analysis uses data for all Tunisian governorates. The main empirical results suggest that water demand is both price and income inelastic, but with high elasticity values (in an absolute sense) estimated for the non-coastal governorates compared to the coastal governorates. Overall, one may argue that combined pricing and non-pricing policies should be implemented in order to save water resources. However, some disparities between Tunisian governorates should be taken into account when applying these policies in order to reach social equity in water consumption. [ABSTRACT FROM AUTHOR]

Water scarcity, climate change and population growth are already stressing the depletion of water supplies. Tunisia like most countries of the Middle East and North Africa region is experiencing severe water shortage problems. However, its water-use sectors are multiple and sometimes conflicting. This may call into question the ability of the current demand management policies to promote further conservation and sustainability of this natural resource. The main purpose of the current study is to shed some lights on the Tunisian water industry focusing mainly on the residential water sector. This latter is the most important use in an urban context because it is straightforwardly related to human survival and hygiene. The descriptive analysis shows, among others, that the residential water industry in Tunisia is far from simple regarding the implemented tariff structure, the involved cross subsidies between consumption blocks, etc. Indeed, despite the improvements in water supply, mobilisation, management and conservation, further efforts should be undertaken to promote the sustainability of the Tunisian water resources. [ABSTRACT FROM PUBLISHER]

Highlights • The extra-intestinal pathovar of E. coli was found to be highly prevalent. • The emergence of the pandemic clone ST131 non-ESBL producer was observed. • qnrS1 resistance gene was found in the intestinal microbiota of Tunisian healthy children. • Intestinal microbiota are important as reservoirs of virulent as well as resistant strains. • Reinforcing hand hygiene and antibiotic rational use are imperative to avoid diffusion of these pathogens in the community. Abstract This study was performed to investigate the distribution of antimicrobial resistance genes and extra-intestinal virulence determinants in a collection of 98 Escherichia coli strains isolated from rectal swabs of healthy children. Forty-six isolated strains were resistant to at least one of the tested antibiotics (usually active against enterobacteria). They were mainly resistant to ampicillin and ticarcillin (42.97%), tetracyclin (26.5%), and trimethoprim/sulfamethoxazole (18.4%). No resistance to the third generation of cephalosporins, carbapenems, aminoglycosides and colistin was found. Resistance to penicillins was encoded by bla TEM-1 (n =34) and bla SHV-1 genes (n =4). Tetracyclin resistance was encoded by tetB (n =12), tetA (n = 5), and tetC (n =1) genes. Amongst resistant quinolones isolated (n =5), chromosomal mutations in gyrA and parC genes were detected in four isolates and qnrS1 gene in two strains. Nine plasmid replicon types were detected; IncFIB (n =36) and IncI1 (n =7) were the most frequent ones. Isolates frequently belonged to phylogenetic groups A (51.1%) and D (27.5%). Extra-intestinal pathovar (n =38) occurred mainly in B2 phylogroup (P =0.0002). Amongst them, two isolates (non-extended-spectrum-β-lactamase (ESBL)-producers) belonged to the pandemic clone ST131. A significant distribution of virulence determinants and pathogenicity island marker was observed within strains belonging to B2 and D phylogroups. Interestingly, our results showed that ExPEC strains, including ST131 pandemic clone, are present within fecal isolates in healthy children. These findings highlight the importance of intestinal microbiota as a reservoir for virulent and resistant strains. Thus, reinforcing hand hygiene and antibiotic rational use is imperative to avoid the diffusion of these pathogens in the community. [ABSTRACT FROM AUTHOR]

Objectives: To describe clinical and molecular characteristics of an outbreak due to metallo-β-lactamases (MBLs) producing Klebsiella pneumoniae collected at Charles Nicolle Hospital of Tunis and to analyze the impact of outer membrane porin (OMP) loss on carbapenem resistance levels. Methods: Between 2010 and 2015, 178 carbapenem-resistant Enterobacteriaceae were isolated. Screening for MBL production was performed using combined disk diffusion method, with imipenem and ethylene diamine tetraacetic acid (EDTA) as inhibitors. Resistance genes and virulence factors were identified by polymerase chain reaction (PCR) and sequencing. Genotyping was performed by pulsed-field gel electrophoresis and multilocus sequence typing. Genetic environment of carbapenemase genes was determined by PCR mapping. Conjugation assays were performed, and plasmids were assigned to incompatibility groups by PCR-based replicon typing. OMPs were profiled by sodium dodecyl sulfate-polyacrilamide gel electrophoresis, and porin genes were sequenced. Results: Nineteen K. pneumoniae (10.6%) showing MBL activity were isolated from patients hospitalized on four different wards. NDM-1 was the only MBL identified, in association with blaOXA-48. All strains lacked at least one OMP, and carbapenem resistance levels were remarkably elevated in strains lacking OmpK35 and OmpK36. blaNDM-1 was located in IncFIA-type conjugative plasmid, with the same genetic context in all strains. The epidemiological diffusion of blaNDM-1 was due to two clones, one major clone belonging to sequence type (ST) 147 (n = 16) and the other clone belonging to ST307 (n = 3). Conclusions: This study describes an outbreak of NDM-1-producing K. pneumoniae strains, isolated from a Tunisian hospital, caused by two clones belonging to ST147 and ST307; and highlights the role of OMPs loss, in combination with β-lactamase expression, in conferring high carbapenem resistance. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is one of the most serious threats to human and animal health. Evidence suggests that the overuse of antimicrobial agents in animal production has led to the emergence and dissemination of multidrug-resistant isolates. The objective of this study was to assess the rate of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in calf feces and to characterize their resistance genes for antibiotics like beta-lactams and colistin, but also to determine their virulence genes. Fecal samples were collected from 100 diarrheic calves in the region of Bizerte, Tunisia. After isolation, E. coli isolates were screened for antimicrobial resistance against 21 antibiotics by the disc diffusion method. Characterization of β-lactamase genes and determination of associated resistance genes were performed by polymerase chain reaction. Among 71 E. coli isolates, 26 (36.6%) strains were ESBL-producing. Most of these isolates were multidrug-resistant (92.3%) and the most prevalent beta-lactamase genes detected were blaCTX-M (n = 26), blaSHV (n = 11), and blaTEM (n = 8), whereas only 1 isolate carried the blaCMY gene. In addition, resistance to carbapenems was detected in two isolates; one of them harbored both blaOXA-48 and blaIMP genes and the other isolate carried only the blaIMP gene. Several resistance genes were identified for the first time in Tunisia from cases of diarrheic calves. Furthermore, to the best of our knowledge, this is the first report of detection and identification of carbapenem resistance genes and virulence genes from calves in North Africa. A high occurrence of antimicrobial resistance of E. coli recovered from fecal samples of calves with diarrhea was observed, highlighting the need for prudent use of antimicrobial agents in veterinary medicine to decrease the incidence of multidrug-resistant bacteria for both animals and humans. [ABSTRACT FROM AUTHOR]

The studies of the HFE mutations: H63D and C282Y in North African populations have revealed the extreme rarity or even the absence of the C282Y mutation. We have examined 1140 chromosomes (570 Tunisian people) for the presence of the two HFE mutations by PCR-RFLP analysis. We have found that the allele frequencies are, respectively, 15.17% (±2.1%) for the H63D and 0.09% (±0.17%) for the C282Y. These results are consistent with the worldwide spread of the H63D mutation and the north European restriction of the C282Y. This study will be completed by determining whether homozygote trait for H63D and associated risk factors (ß thalassémia) can lead to iron overload in Tunisia. [Copyright &y& Elsevier]