Epidemiology of Campylobacter jejuni and Campylobacter coli Infections in the Zenica – Doboj Canton, Bosnia and Herzegovina – A Laboratory Based Surveillance in the 1999–2001 Period

Previous studies in the Zenica – Doboj Canton, Bosnia and Herzegovina, indicated some different epidemiological features of Campylobacter infections and high degree of antimicrobial resistance. Therefore, it was important to investigate epidemiology of Campylobacter jejuni and Campylobacter coli infections by demographic features and antimicrobial resistance in the 1999–2001 period. A total number of 40 (75.5%) C. jejuni and 13 (24.5%) C. coli non-repeated clinical isolates were analyzed. More than half of isolates, 30 (56.6%) were from urban dwellers. Campylobacter isolates mainly obtained from children under 6 years of age, 42 (79.2%), resulting in far off highest incidence rate of 41.4/100,000/year in this age group. There was noted high degree of resistance to ciprofloxacin in children less than 6 years of age (14.3%), and extremely high overall erythromycin-resistance rate (30%). Campylobacteriosis in this region is a public health concern not in the term of the number reported cases, but of distinctive epidemiologic features

Antibiotic resistance in Enterobacter cloacae strains with derepressed/partly derepressed/inducible AmpC and extended spectrum beta-lactamases in Zenica-Doboj Canton, Bosnia and Herzegovina

AIM: To investigate the prevalence of derepressed/partly derepressed/inducible and ESBL/AmpC-producing Enterobacter cloacae isolates and treatment options for infections associated with those isolates. ----- METHODS: Antibiotic susceptibility was determined by disc diffusion and broth microdilution according to CLSI guidelines. Doubledisk synergy test (DDST) was performed in order to screen for ESBLs and combined disk test with phenylboronic acid to detect AmpC β -lactamases. PCR was used to detect blaESBL/blacarb genes. Genetic relatedness of the strains was determined by pulsed-fieldgel-electrophoresis (PFGE). ----- RESULTS: Among 14 isolates with the ESBL positive E. cloaceae producing isolates, four (28.6%), nine (64.3%) and one (7.1%) isolates were derepressed/partly derepressed and inducible AmpC producers. Eleven (out of 14) isolates were resistant to cefotaxime, ceftazidime, ceftriaxone, aminoglycosides and fluoroquinolones. All isolates were susceptible to imipenem and meropenem, 79% to cefepime. Five (out of 14; 35.7%) isolates (four derepressed and one inducible AmpC carrying E. cloaceae) were negative in phenotypic test for ESBLs, but positive for broad spectrum TEM-1 β-lactamase. One (out of four derepressed) also produced CMY-2 β-lactamase. Four (out of nine) partly derepressed isolates were positive with the DDST, but did not yield PCR products with primers targeting TEM, SHV and CTX-M beta-lactamases. Four positive partly derepressed isolates carried a blaCTX-M-1 gene, two blaOXA-1 one blaCTX-M-15, OXA-1 and one blaCTX-M-28, OXA-1 (n=1). ----- CONCLUSION: Microbiology laboratories must be able to detect and recognize AmpC-carrying isolates in a timely manner, especially those that are falsely susceptible in vitro to drugs that may be consideredfor therapy of infected patients

Prevalence of co-existence genes and clonal spread of ESBL-producing isolates causing hospital and community-acquired infections in Zenica-Doboj Canton, Bosnia and Herzegovina

Introduction: Co-existence type of ESBL-producing isolates are serious problem in the public health world. Methods Antibiotic susceptibility was determined by disc diffusion and broth microdilution according to CLSI guidelines. Double-disk synergy test was performed in order to screen for ESBLs/pAmpC beta-lactamases. PCR was used to detect blaESBL/blaampC/blacarb genes. Genetic relatedness of the strains was determined by pulsed-field-gel-electrophoresis (PFGE). Results In this study 88 of the inpatient isolates (n=126; 10.0%) and 62 of the outpatient (n=184; 6.4%) Beta-lactamase-producing isolates were taken for the study. They included 50.0/29.0% K. pneumoniae, 12.5/30.6% E. coli, 11.4/4.8% A. baumannii, 8.0/14.5% K. oxytoca, 8.0/4.8% E. cloacae, 5.7/8.1% Proteus spp., and less than 3.5% of other isolates. Co-existence of more than two type of beta-lactamases was detected in 77.3% of inpatient and 45.2% of outpatient isolates. Among inpatient isolates, Klebsiella spp. and E. coli were the most frequent isolates which produce more than two type of genes; in ≈ 65% and ≈12% cases. Separately, combination of four: TEM+SHV+CTX-M+OXA-1 beta-lactamases in inpatient K. pneumoniae isolates were detected in 63.6% cases respectively. Differents in antimicrobial resistance were higher to cephalosporins agents in Klebsiella spp., and E. coli at inpatient and outpatient isolates which produce more than two types of beta-lactamases than in isolates which produce one type of beta-lactamases. Conclusion: This work demonstrates a progressively increasing prevalence of co-existence type of beta-lactamases expecially in inpatient isolates. Continous monitoring and surveillance and proper infection control and prevention practice will limit the further spread of these isolates

Amino Acids Composition and Antioxidant Activity of Selected Mushrooms from Bosnia and Herzegovina

Many studies highlight the health benefits of mushrooms, which are consequently becoming more and more interesting for researchers. The content of amino acids (AA), total phenolic content (TPC), and antioxidative activity (AAc) were determined in wild as well as cultivated mushrooms. The AA included: L-tryptophan (Trp), L-arginine (Arg), L-cysteine (Cys), L-methionine (Met), L-alanine (Ala), L-phenylalanine (Phe), L-lysine (Lys), L-valine (Val), L-glycine (Gly), and L-leucine (Leu). The wild mushrooms: Lactarius piperatus, Amanita caesarea, Lactarius deliciosus, Lycoperdon pyriforme, Macrolepiota procera, and cultivated mushrooms: Agaricus bisporus, Boletus aestivalis, Cantharellus cibarius, Pleurotus ostreatus, and Agaricus bisporus var. avellaneus were investigated in this study. The AA was determined by HPTLC method and quantified with a Camag TLC scanner and WINCAT software by scanning the plates at 540 nm. The TPC was spectrophotometrically estimated as gallic acid equivalents/g of fresh weight according to Folin-Ciocalteu’s method. The radical scavenging activity (RSA) of mushroom extracts was determined by DPPH assay. The highest content of Ala, Gly, Phe, Lys, Val, and Leu was found in Pleurotus ostreatus. The total phenolic content (mg GAE/g) in investigated mushrooms ranged from 1.90 to 35.56, and the % RSA ranged from 43.88 to 90.17. This study promotes the consumption of food rich in bioactive compounds, mushrooms being among such food. Further research on mushrooms from Bosnia and Herzegovina and their benefits in the overall maintenance of human health and protection from age-related diseases is necessary

Arrival of carbapenem-hydrolyzing-oxacillinases in Acinetobacter baumannii in Bosnia and Herzegovina

Multidrug-resistant Acinetobacter baumannii (MDR-AB) is an important threat for critically ill patients. It can infect the respiratory tract, blood, soft tissues, urinary tract and central nervous system. Recently, carbapenem-resistance was observed in A. baumannii clinical isolates from Bosnia and Herzegovina. This prompted us to analyze these isolates with regards to genotypic diversity, antibiotic susceptibility and occurrence of acquired carbapenem resistance genes. Twelve carbapenem-resistant isolates were collected at a University hospital during two different periods of 2011 and 2015-2016: four isolates in 2011 and eight isolates 2015-2016 and compared to determine the dynamic changes in carbapenemase resistance mechanisms and population structure. All twelve isolates were positive for intrinsic blaOXA-51-like, nine for blaOXA-40-like and one for the blaOXA-23-like gene. ISAba1 was found upstream of blaOXA-51 in all and upstream of blaOXA-23-like gene in one isolate. Sequencing of the selected PCR products revealed the presence of OXA-72 β-\ud lactamase (strain 1) and OXA-23 β-lactamase (strain 41). WGS of the selected isolate (AB 5) revealed the presence of blaOXA-72, chromosomal genes blaOXA-69 and blaADC. Moreover, the aac (3)-1a and aadA1 genes encoding aminoglycoside resistance, and sul1encoding sulphonamide resistance were identified. PFGE and rep-PCR revealed two clones containing highly similar isolates positive for OXA-40-like; one from 2011 and the other from 2015- 2016. Implementation of hospital hygiene measures, screening of the patients on admission for carriage of MDR-AB, and the early and accurate detection, with restriction of antibiotic use should be recommended to control the spread of these important hospital pathogenes. To our knowledge, this is the first report of A. baumannii isolates producing carbapenem-hydrolyzing oxacillinases (CHDL) from Bosnia and Herzegovina

Molecular epidemiology and antimicrobial susceptibility of AmpC- and/or extended-spectrum (ESBL) ß-lactamaseproducing Proteus spp. clinical isolates in Zenica-Doboj Canton, Bosnia and Herzegovina

Aim To investigate prevalence, antimicrobial susceptibility, molecular characteristics, and genetic relationship of AmpC- and/or extended spectrum beta lactamase (ESBL)- producing Proteus spp. clinical isolates in Zenica-Doboj Canton, Bosnia and Herzegovina. Methods Antibiotic susceptibility was determined by disc diffusion and broth microdilution methods according to CLSI guidelines. Double-disk synergy test was performed in order to screen for ESBLs, and combined disk test with phenylboronic acid to detect AmpC β -lactamases. PCR was used to detect blaESBL/blacarb genes. Genetic relatedness of the strains was determined by pulsed-fieldgel-electrophoresis (PFGE). Results Eleven ESBL-producing isolates were included in the study (six inpatients and five outpatients). Susceptibility rate to amoxicillin-clavulanic acid, imipenem and meropenem was 100%. Resistance rate to cefuroxime was 100%, gentamicine 90.9%, piperacillin/tazobactam 81.8%, cefotaxim, ceftriaxone and ceftazidime 72.7%, cefoxitine and ciprofloxacine 63.6% and to cefepime 45.5%. In five (out of 11) isolates multi-drug resistance (MDR) to cephalosporins, cefamicines, amynocligosides and fluoroquinolones was detected. Besides TEM-1 which was detected in all isolates, CTX-M+OXA-1 β-lactamases were detected in seven (out of 11; 63.6%) isolates (five blaCTX-M-1 and two blaCTX-M-15 genes), and CMY-2 β-lactamase in two isolates. PFGE showed no genetic relatedness. Conclusion Because of high prevalence of MDR strains in epidemiologically unrelated patients with AmpC- and/or ESBL producing Proteus spp. infection, further surveillance is needed. Molecular characterization and strain typing, or at least phenotypic test for AmpC/ESBL production is important for appropriate therapy and the detection of sources and modes of spread, which is the main step in order to design targeted infection control strategies