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Abstract
Ventilator-associated pneumonia (VAP) is a serious and common complication in patients admitted to intensive care unit (ICU) and contributes to mortality. Multidrug Gram-negative bacteria such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae are frequently associated with VAP in ICU. A prospective study was set up in three ICUs of the University Hospital Center Zagreb and one ICU in General Hospital Pula from September 2017 to March 2018. Antibiotic susceptibility was determined by broth microdilution method. Production of extended-spectrum β-lactamases (ESBLs) was determined by double-disk synergy test and carbapenemases by Hodge and carbapenem inactivation method (CIM). The genes encoding ESBLs, carbapenemases of class A, B and D and qnr genes were determined by PCR. In total 97 Gram-negative bacteria isolates were analyzed. P. aeruginosa demonstrated high resistance rates for imipenem and meropenem with 74% and 68% of resistant strains, respectively. Moderate resistance rates were observed for ceftazidime andpiperacillin/tazobactam, ciprofloxacin and gentamicin (44%). All except three A. baumannii isolates, were resistant to carbapenems and to all other antibiotics apart from colistin and amikacin. Eight A. baumannii isolates were positive for blaOXA-23 and 12 for blaOXA-24 genes. Four K. pneumoniae and two E. cloacae strains were ESBL positive and harboured group 1 of CTX-M β-lactamases. Three P. mirabilis strains were positive for plasmid-mediated ampC β-lactamase of CMY family. Two carbapenem-resistant K. pneumoniae harboured OXA-48 and one carbapenem-resistant E. cloacae VIM-1. A high proportion of multidrug-resistant P. aeruginosa, K. pneumoniae and extensively resistant A. baumannii was reported. Acquired resistance mechanisms, mainly production of carbapenemases and ESBLs were dominant in A. baumannii and K. pneumoniae, respectively. Resistance of P. aeruginosa isolates was more likely due to upregulation of efflux pumps or porin loss. A marked diversity of β-lactamases was identified in Enterobacteriaceae.
Conflict of interest
Authors: Daniela Bandić-Pavlović, Tajana Zah-Bogović T, Marta Žižek, Luka Bielen, Vesna Bratić, Pero Hrabač, Slobodan Mihaljević, Domagoj Slačanac, and Branka Bedenić declare no conflict of interest.
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Notes on contributors
Daniela Bandić-Pavlović
Daniela Bandić-Pavlović, MD, PhD, Assistant professor works School of Medicine, University of Zagreb, University Hospital Center Zagreb. She is the leader of the elective course and I participate in other elective courses. She also participates in the postgraduate specialist study of Anesthesiology, Reanimation and Intensive Medicine (where she is also a leader of one subject) and General Surgery. She is the leader of one postgraduate study of the first category, and I participate in many other postgraduate studies of the first category. Her scientific interest are anestesiology, intensive, care, infections in ICU.
Tajana Zah-Bogović
Tajana Zah Bogović, MD, PhD. Worked at University Hospital Center Zagreb, School of Medicine, University of Zagreb as assistant professor. She is specialist in anestesiology and intensive care. Tajana Zah Bogović, was born on January 30th 1977 in Zagreb. She enrolled at the Faculty of Medicine, University of Zagreb in 1996, and graduated in 2002. In 2003, she passed the professional exam for doctors of medicine. Since 2004 she has been working at the Clinical Hospital Center Zagreb at the Clinic of Anesthesiology, Resuscitation and Intensive Care as a resident, then a specialist and subspecialist. She passed the specialist exam in 2008, and the subspecialist exam in the specialty of intensive care medicine in 2013. In 2014 she defended her doctoral dissertation entitled “Transfusion-induced acute lung injury (TRALI): Properties and frequency in the Republic of Croatia”. She attended professional training in the USA (Cincinnati Children's), Austria and Italy. She is the author and co-author of a number of scientific and professional published papers. From 2008 to 2018 she has been actively participating in teaching and researching at the Medical School, University of Zagreb, field anesthesiology and intensive care. From September 2018. she is working and living in Austria. Her scientific interest are infections in ICU.
Marta Žižek
Marta Žižek, PhD, molecular biologist, Associate Researcher is employed at the University of Zagreb, Faculty of Sciences. She participated in FAPIC in FAPIC (http://www.fapic.eu.) in validation of a fast and cost-effective test for the diagnosis of sepsis. She was associate researcher in Nanjing, Jiangsu, China from Apr 2019 – Dec 2019, As a member of Microbiology research group, she was working on a research project towards PhD in the areas of Environmental Health and Aerobiology. The main outcome was development of a pipeline for temporal and spatial analysis of propagules in the air. She has constructed the robotic impact type samplers that were deployed to collect the air samples. Gained skills in electronics and physics, like soldering and electrical equipment testing. She was in the PhD programme in Aberystwyth University UK from 2013 to 2018. Her scientific interests are molecular microbiology, mycology, environmental microbiology.
Luka Bielen
Luka Bielen, MD, PhD, assistant professor, internist, is empoyed at Medical Faculty, University of Zagreb, University Hospital Center Zagreb at the Deparment for Intensive Care Medicine. He was in 2014 a collaborator, project “Prospective randomized clinical trial: efficacy of fosfomycin vs. ertapenem in the treatment of urinary tract infections caused by extended spectrum β-lactamase producing strains of E. coli”. His scientific interest are clinical pharmacology and antibiotic chemotherapy. His scientific interest are antimicrobial chemotherapy and resistance.
Vesna Bratić
Vesna Bratić is employed at the University Hospital Center Zagreb, Department for Anesteziology and intensive care of the University Hospital Center Zagreb. E. Chief nurse at the Department for Anesteziology and intensive care of the University Hospital Center Zagreb. Enrolled at the PhD programme at the University of Zagreb Dental School.
Vesna Bratić, graduated in 2013 from the Medical Faculty of the University of Zagreb with a master's degree in nursing. From 1995 to 2003 she worked at the University Hospital Center Zagreb, as a nurse in the Intensive Care Unit of Surgical Patients, from 2004 to 2006 as a nurse in anesthesia. From 2006 to 2019 she worked as the head nurse of the Department of Anesthesiology and intensive treatment of surgical and urological patients, and from 2020 in the position of head nurse of the Clinic for Anesthesiology, Resuscitation and Intensive Care. 2008 he is elected to the associate title of assistant as the title title at the Department of Health Care at the Medical Polytechnic in Zagreb. She was training at St George's Healthcare NHS, London, England, in 2011. She has published a number of professional and scientific papers at home and abroad, and is currently working on chapters in the book related to anesthesiology and intensive care. Her scientific interest are infections in ICU.
Pero Hrabač
Pero Hrabać, MD, PhD employed at University of Zagreb, School of Medicine. Graduated from the University of Zagreb School of Medicine. Enrolled in the PhD Programme “Biomedicine and Health”. He worked in the pharmaceutical industry: – Boehringer Ingelheim, Pfizer, Lek, JGL, Belupo, CSC Pharmaceuticals, Cyathus. Participated in the multicenter clinical studies. Experienced in the organization of the informatic support for the clinical investigation of the new drugs. Since 2011 CARNET systemic ingener for the University of Zagreb School of Medicine. Since 2014 assistant at the Department for informatics, statistics and epidemiology. His scientific interests are medical informatics, medical statistics.
Domagoj Slačanac
Domagoj Slačanac, MD, PhD is employed at the Hospital Düsseldorf, North Rhine-Westphalia, Germany. In 2012, he enrolled in the study of medicine at the Faculty of Medicine, University of Zagreb. During my Studies he worked as a Student Teaching Assistant at the Departments for Microbiology and Biochemistry, as well as contibuted to published scientific studies in the fields of Bacteriology and Virology at the Microbiology department of the School of Medicine in Zagreb. In April 2019 he acquired his License to practice Medicine and since then I has worked as an Assistant Physician in the field of Epidemiology in a Public Health Department and Psychiatry in a Maximum care Hospital. Currently he is involved as a Physician in the Biomonitoring of the CoV-19 Pandemic in the city of Cologne. His scientific interest are : antimicrobial resistance, mycology.
Slobodan Mihaljević
Prof. dr. sc. Slobodan Mihaljevic, MD, PhD is Head of the Department of Anesthesiology and Intensive Care Unit at the University Hospital Centre Zagreb. In over 35 years of clinical experience, he has specialized in anaesthesiology, emergency medicine and intensive care. In addition, he holds teaching positions in anaesthesiology at University of Zagreb and University of Mostar. His research interests include the latest advancements in the field.
Branka Bedenić
Branka Bedenić, MD, PhD, full professor tenure, employed at University of Zagreb School of medicine, University Hospital Center Zagreb. Since 2017 Full professor tenure, Department for microbiology and parasitology, School of Medicine. She participated in many projects on antimicrobial resistance and is collaborator at the Horizon project FAPIC (Fast assay for pathogen identification and characterization). She published over 90 papers in established international journals and is leader of many subjects for undergraduate and postgraduate students. She was a visiting researcher in many established Institutions abroad. She supervised 10 PhD Thesis and 15 Bachelor degree Thesis. Her scientific interests are beta-lactamases, carbapenemases and antimicrobial resistance.