Abstract Extended-spectrum β-lactamases (ESBLs) hydrolyse extended-spectrum cephalosporins (ESC) and aztreonam. As ESBL-producing organisms have been identified in food producing animals, the aim of our study was to detect and analyse such Escherichia coli isolates from poultry. Antibiotic susceptibility of the isolates was determined with disk-diffusion and broth microdilution methods. ESBLs were detected with the double-disk synergy and inhibitor-based test with clavulanic acid. The transferability of cefotaxime resistance was determined with conjugation experiments, and genes encoding ESBLs, plasmid-mediated AmpC β-lactamases, and quinolone resistance determinants identified by polymerase chain reaction. The study included 108 faecal samples (cloacal swabs) from 25 different poultry farms in the Zenica-Doboj Canton, Bosnia and Herzegovina. Of these, 75 (69.4 %) were positive for E. coli, of which 27 were resistant to cefotaxime, amoxicillin, cefazoline, and cefriaxone, and susceptible to imipenem, meropenem, ertapenem, and amikacin. All 27 cefotaxime-resistant isolates were positive in double-disk synergy and combined disk tests. Eighteen isolates transferred cefotaxime resistance to E. coli recipient. Twenty-one isolates were positive for the blaCTX-M-1 cluster genes and seven for blaCTX-M-15. Fourteen were positive for the blaTEM genes. The most frequent plasmid incompatibility group was IncFIB, whereas IncFIA and Inc HI1 were present in only a few isolates. Two different sequence types (STs) were identified: ST117 and ST155. The emergence of ESBL-producing E. coli in farm animals presents a public health threat, as they can colonise the intestine and cause infections in humans.

Department of Clinical Microbiology, Institute for Health and Food Safety Zenica, Fra Ivana Jukića 2, 72000 Zenica, Bosnia and Herzegovina Department of Epidemiology, Institute for Health and Food Safety Zenica, Fra Ivana Jukića 2, 72000 Zenica, Bosnia and Herzegovina Autoimmune Diseases Department, Vasculitis Research Unit, IDIBAPS, CELLEX 4B, Hospital Clinic Barcelona, Spain Department of Chemistry Diagnostics, Institute for Health and Food Safety Zenica, Fra Ivana Jukića 2, 72000 Zenica, Bosnia and Herzegovina Department for Epizootiology, Institute for Health and Food Safety Zenica, Fra Ivana Jukića 2, 72000 Zenica, Bosnia and Herzegovina

Aim The damage caused by the COVID-19 pandemic has made the prevention of its further spread at the top of the list of priorities of many governments and state institutions responsible for health and civil protection around the world. This prevention implies an effective system of epidemiological surveillance and the application of timely and effective control measures. This research focuses on the application of techniques for modelling and geovisualization of epidemic data with the aim of simple and fast communication of analytical results via geoportal. Methods The paper describes the approach applied through the project of establishing the epidemiological location-intelligence system for monitoring the effectiveness of control measures in preventing the spread of COVID-19 in Bosnia and Herzegovina. Results Epidemic data were processed and the results related to spatio-temporal analysis of the infection spread were presented by compartmental epidemic model, reproduction number R, epi-curve diagrams as well as choropleth maps for different levels of administrative units. Geovisualization of epidemic data enabled the release of numerous information from described models and indicators, providing easier visual communication of the spread of the disease and better recognition of its trend. Conclusion The approach involves the simultaneous application of epidemic models and epidemic data geovisualization, which allows a simple and rapid evaluation of the epidemic situation and the effects of control measures. This contributes to more informative decision-making related to control measures by suggesting their selective application at the local level.

Objectives: To determine the epidemiological features of patients and animals after bites/scratches from rabies-suspected animals in Zenica-Doboj Canton, Bosnia and Herzegovina. Methods: Data from all patients (and the causative animals) admitted to the Antirabies Service of the Institute for Health and Food Safety Zenica in the 2009-2017 period were analyzed, including age, sex, anatomical site of the bite/scratch, animal type (stray/owned/wildlife), veterinary observations of the animal, and whether antirabies post-exposure prophylaxis (PEP) was indicated and/or administered. Results: In total, 1716 patients were admitted. Bites/scratches were most frequently recorded during April and May (n=181, 10.5% and n=163, 9.5%, respectively). The persons admitted were mostly from the Zenica municipality (n=1278, 74.5%; incidence: 11.55/ 1000), which is 66.6% urbanized. Males were more frequently represented (n=1089, 63.6%). The patients were mostly 50-64 and 25-49 years of age (n=425, 24.7% and n=390, 22.7%, respectively). Dog bites were the most common cause (n=1634, 95.1%, of which n=1258, 77.0% were caused by stray dogs). PEP was indicated for 997 (58.1%) patients. Only 340 (19.9%) animals underwent veterinary observations (3.1% of stray and 76.1% of owned animals). The largest number of injuries were presented at lower extremities, 1044 (60.8%) cases. Conclusions: Zenica-Doboj Canton is a rabies-free region. Due to the high rate of stray animals not undergoing veterinary observations, the non-existence of a unique dog registry, and the consequent lack of information about stray animals in terms of number, vaccination, neutering, and euthanasia, there is an urgent need for improving the prevention and control of rabies within the One Health framework.

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 bla OXA-51-like , nine for bla OXA-40-like and one for the bla OXA-23-like gene. IS Aba1 was found upstream of bla OXA-51 in all and upstream of bla OXA-23-like gene in one isolate. Sequencing of the selected PCR products revealed the presence of OXA-72 β-lactamase (strain 1) and OXA-23 β-lactamase (strain 41). WGS of the selected isolate (AB 5) revealed the presence of bla OXA-72 , chromosomal genes bla OXA-69 and bla ADC . Moreover, the aac (3 )-1a and aadA1 genes encoding aminoglycoside resistance, and sul1 encoding 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.

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.

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.

BACKGROUND/AIM The aim of this study was to determine the molecular characteristics and antibiotic resistance of 13 (10 inpatient and three outpatient) Acinetobacter baumannii beta-lactamase-producing isolates collected in Bosnia and Herzegovina between December 2009 and May 2010. MATERIALS AND METHODS Susceptibility testing was performed by disk diffusion and broth microdilution methods. The modified Hodge and combined disk test with EDTA/phenylboronic acid was used to screen for carbapenemase production. Production of extended spectrum beta-lactamases (ESBLs) was determined by double-disk synergy test. PCR was used to detect blaESBL/blacarb genes. RESULTS Ten (22.2%) inpatient and three (13.6%) outpatient isolates produced beta-lactamases, ESBLs, or oxacillinases. More than 50% of the isolates showed multidrug resistance. Resistance rates to gentamicin and ciprofloxacin of the inpatients and outpatients were 80.0%, 60.0%, 75.0%, and 25.0%, respectively. MICs of carbapenems for resistant isolates ranged from 32 to >256 μg/mL. All imipenem-resistant Acinetobacter baumannii strains contained blaOXA-51. Three of the 10 inpatient isolates and one outpatient isolate containing blaOXA-51 additionally produced other beta-lactamases (TEM/CTX-M/OXA-1). None of the inpatient or outpatient isolates were positive for other carbapenemases, especially acquired oxacillinases (blaOXA-23/blaOXA-24/blaOXA-58/blaOXA-143). CONCLUSION Production of blaOXA-51 presents an emerging threat in imipenem-resistant Acinetobacter spp. from Bosnia and Herzegovina.