Aim To characterize methicillin-resistant S. aureus (MRSA) strains phenotypically and genotypically and to determine their clonal affiliation, representation and antibiotic resistance profile. Methods A total of 62 randomly selected MRSA isolates of different clinical samples collected from 2009 to 2017 were phenotypically and genotypically analysed. Phenotypic analyses were performed by standard microbiological procedures, and using VITEK 2/AES instrument as well as MALDI-TOF (matrix-assisted laser desorption/ionization) technology. Genotypic characterization included spa, MLST (multilocus sequence typing) and SCCmec typing, and detection of the Panton-Valentine leukocidin (PVL) and other enterotoxin encoding genes. Results The largest number of isolates, 21 (33.87%) belonged to ST228-MRSA-I, spa type t041, t1003 and t001. Other major clones were: ST239-MRSA-III, spa type t037 and t030 (27.41%); ST8-MRSA-IV, spa type t008 and t121 (12.9%); ST247-MRSA-I, spa type t051 (4.83%). PVL was detected in 10 isolates (SCCmec IV/V). During 2009 and 2010 the most frequent MRSA strain was South German clone, ST228-MRSA-I (80% and 90%, respectively), while in later years it was replaced with Brazilian-Hungarian clone ST239-MRSA-III (75% in 2015 and 2016). The South German clone, spa type t041 in 90.48% of cases was resistant to clindamycin, ciprofloxacin, erythromycin, cefoxitin, gentamicin, kanamycin, tobramycin and penicillin, while 70.58% samples of the Brazilian-Hungarian clone spa type t037 were additionally resistant to tetracycline and rifampicin. Conclusion This research can supplement the existing knowledge about the clonal distribution of MRSA in Bosnia and Herzegovina and their sensitivity to antibiotics in order to improve the national control of these infections.

Introduction: The most common cause of myocardial ischemia is atherosclerotic epicardial coronary artery disease, present in 90% of patients. Risk factors positively correlate with the onset, development and subsequent complications of atherosclerotic disease. Aim: Determine the percentage frequency of classic risk factors for coronary disease in patients with non-ST segment elevation myocardial infarction (NSTEMI), with regard to gender. Methods: A retrospective study was conducted on 600 respondents, treated for NSTEMI at the Clinic for Internal Medicine of the University Clinical Center (UKC) Tuzla, in the period from June 2016 to December 2019. Results: Overall, smoking was the leading risk factor (65%), followed by hypertension (58%), hyperlipoproteinemia (39%), overweight (33%), positive family burden (30%) and diabetes mellitus (19%). In male patients, the leading risk factor was smoking, rating at 74%, while in female patients – it was hypertension at 67%. In younger groups of patients leading risk factors were smoking and a positive family burden. Conclusion: With adequate prevention and treatment measures, a significant reduction in the prevalence of the cardiovascular disease can be achieved, since the risk factors for its development have long been known. Quitting smoking is one of the most effective secondary prevention measure since it reduces the reinfarction risk rate by 50%. Knowledge of coronary risks, as well as success in reducing them, can greatly contribute to patients’ overall sense of contentment and significantly raise their self-confidence.

Aim: Nosocomial infections (NIs) represent a major public health problem in developed, and even more in developing countries. Based on the origin of the pathogen, NIs are divided into endogenous caused by microorganisms of the patient’s microflora, and exogenous caused by pathogens from the patient’s environment or by the same microorganism isolated elsewhere from patient’s body. The main aim of this research was to determine the frequency, etiology and types of NIs at the Clinic for Surgery of the University Clinical Center Tuzla. Material and Methods: 5.039 patients were prospectively followed for the development of endogenous and exogenous NIs (January-December 2015). The definition of NI was performed using standardized the National Healthcare Safety Network (NHSN) criteria. Results: Based on continuous epidemiological surveillance, the incidence of NI was 3.51%; with a total of 177 registered infections, 24 endogenous and 153 exogenous. The most common NIs were urinary tract infections (UTIs) (14.29% endogenous and 85.71% exogenous) and surgical site infections (SSIs) (16.67% endogenous and 83.33% exogenous), p<0.001. Gram-negative bacteria were predominant (76.84%) over Gram-positive bacteria (23.16%). Gram-positive bacteria mainly caused bacteremia, while Gram-negative bacteria were most commonly isolated from UTIs and SSIs. The most common Gram-negative bacteria were Klebsiella pneumoniae (24.86%) and Pseudomonas aeruginosa (22.6%), and among Gram-positive, Staphylococcus aureus (10.73%) and coagulase-negative staphylococci (7.91%). Conclusion: Epidemiological surveillance is considered a key link in the program for the prevention and control of NIs. The most important, and the ultimate goal and purpose of conducting epidemiological surveillance are to reduce and eliminate the risk factors, which can lead to a reduction in NI incidence rate. Determining the endemic rates of NIs provides an objective understanding of the overall NI status in an institution as well as existing risk factors for the occurrence of these infections.

Background: Colonization is the presence of bacteria in the intestines, skin, nose, throat or anywhere in the human body without any signs of infection but with increased risk for spreading bacteria to other patients and the emergence of new infections. Screening of colonized patients is used as part of the prevention and control of multidrug-resistant (MDR) infections caused by agents such as methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis and multi-resistant Gram-negative bacteria. Material and methods: Data from 169 screened respondents hospitalized at the University Clinical Center Tuzla, between October 1, 2018, and May 1, 2019, were analyzed. Swabs were taken from nostrils, throat, axilla and groin area for all patients, and also from the anorectal area for 157 of them. Identification of MDR bacteria was done by phenotypical methods, according to the recommendations of EUCAST Clinical Breakpoint Table v.8.0, 2018. Results: Out of 169 patients, negative screening test results were found in 93 examinees (55.02%), and positive in 76 (44.97%). The largest number of patients undergoing screening was hospitalized in the Intensive Care Unit (Surgical Block), 73/169 (43.19%). The average screening time was 2.2 days or 53 hours. In 18/76 (23.68%) of patients with positive screening, prior to screening regular microbiological testing was done, and in the remaining 58/76 (76.3%) screening was performed first. Analyzing respondents with positive screening, 27 (35.53%) had negative findings during regular microbiological testing of different biological samples and for 49 (64.47%) different/same strains of MDR bacteria were isolated.Conclusion: Knowing the phenotypic profile of bacteria colonizing patients in intensive care units is a very useful tool in preventing their spread intra- and inter-hospitals.

Aim To determine the prevalence rate and resistance profile of Streptococcus agalactiae (S. agalactiae) in vaginal swabs of pregnant and adult non-pregnant women in the Tuzla region, Bosnia and Herzegovina (B&H), as well as its association with other aerobic bacteria. Methods This prospective study included 200 women, 100 pregnant and 100 adult non-pregnant. The research was conducted at the Institute of Microbiology, University Clinical Center Tuzla from October to December 2015. Standard aerobic microbiological techniques were used for isolation and identification of S. agalactiae and other aerobic bacteria. Antimicrobial susceptibility was determined by the disk diffusion and microdilution method(VITEK 2/AES instrument). Results Among 200 vaginal swabs, 17 (8.50%) were positive for S. agalactiae, e. g., 7% (7/100) of pregnant and 10% (10/100) of adult non-pregnant women. In the pregnant group, 71.4% (5/7) of S. agalactiae isolates were susceptible to clindamycin and 85.7%(6/7) to erythromycin. In the adult non-pregnant group, only resistance to clindamycin was observed in one patient (1/10; 10%). S. agalactiae as single pathogen was isolated in 57.14% (4/7) of pregnant and 60% (6/10) of adult non-pregnant S. agalactiae positive women. In mixed microbial cultures S. agalactiae was most frequently associated with Enterococcus faecalis and Escherichia coli. Conclusion The rate of S. agalactiae positive women in the population of pregnant and adult non-pregnant women of Tuzla Canton, B&H is comparable with other European countries. Large studies are needed to develop a common national strategy for the prevention of S. agalactiae infection in B&H, especially during pregnancy.

Introduction: Acinetobacter baumannii is a frequent cause of infections in hospitals around the world, which is very difficult to control and treat. It is particularly prevalent in intensive care wards. Aim: The main objective of the research was to establish the application of epidemiological monitoring of nosocomial infections (NIs) caused by A. baumannii in order to determine: the type and distribution of NIs, and to investigate antimicrobial drug resistance of A. baumannii. Material and Methods: 855 patients treated at the Clinic of Anesthesiology and Reanimation, University Clinical Center Tuzla during 2013 were followed prospectively for the development of NIs. Infections caused by A. baumannii were characterized by the anatomical site and antibiotics resistance profile. Results: NIs were registered in 105 patients (12.3%; 855/105). The predominant cause of infection was A. baumannii with an incidence of 51.4% (54/105), followed by ESBL-producing Klebsiella pneumoniae with 15.2% (16/105) of cases, methicillin-resistant Staphylococcus aureus with 8.6% (9/105), and ESBL-producing Proteus mirabilis with 7.6% (8/105). According to the anatomical site, and type of NIs caused by A. baumannii, the most frequent were respiratory infections (74.1%; 40/54). Infections of surgical sites were registered in 11.1% (6/54) of cases, while bloodstream infections in 9.2% (5/54). A. baumannii isolates tested resistant against most antibiotics examined, but showed a high degree of susceptibility to tobramycin (87%; 47/54) and colistin (100%; 54/54). Conclusion: The increasing incidence of multi- and extensively drug-resistant Acinetobacter spp. emphasizes the importance of administration of an adequate antibiotic strategy and the implementation of strict monitoring of the measures for controlling nosocomial infections.

Introduction: Intensive care units (ICUs) are associated with a greater risk of developing nosocomial infections (NIs) than other departments. Aim: The aim of this study was to determine the rate, the site and causative organisms of NIs in the surgical ICU at University Clinical Center Tuzla. Methods: All patients admitted to the surgical ICU were followed prospectively, for the development of NIs (January-December 2010). Determination of NIs was performed using standardized the Centers for Disease Control and Prevention (CDC) criteria. Results: 94 out of 834 patients (11.27%) developed NIs. Respiratory tract infections were seen in 56 (60%), urinary tract infections in 15 (16%) and gastrointestinal tract infections in 8 (9%) patients. Other infections identified were surgical site, bloodstream and skin infections. Gram-negative organisms were reported in approximately 75% of cases (78.7% extended-spectrum beta-lactamase (ESBL)-producers). Klebsiella pneumoniae was the commonest (51.0%), followed by Proteus mirabilis (21.3%) and Pseudomonas aeruginosa (10.6%). Methicillin-resistant Staphylococcus aureus (MRSA) (16%), and Clostridium difficile (9.6%) were the commonest among gram-positive bacteria. Conclusion: Respiratory and urinary tract infections made up the great majority of NIs. ICU patients are more susceptible to NIs, emphasizing the importance of continuous surveillance and enforcement of specific infection control measures.

Introduction: Clostridium difficile (C. difficile) is currently the leading cause of healthcare-associated diarrhea, but almost nothing is known about the extent of C. difficile infection (CDI) in Bosnia and Herzegovina. Goal: We aimed to retrospectively analyze CDI in hospitalized patients at University Clinical Center (UCC) Tuzla, Bosnia and Herzegovina from January 2009 through June 2012. Methods: We analyzed all patients (except children ages 0-2), diagnosed with CDI based on anamnestic and epidemiological, clinical picture and microbiological tests (proof of toxins in the stool by enzyme-linked immunosorbent assay). Results: From a total of 989 patients tested for C. difficile toxin (60.2 per 10,000 inpatient days) 347 (35.08%) were positives. The mean incidence rate of CDI was 2.23 per 10,000 inpatient days (range 1.32-2.87). Annual rates of hospitalization were 15.68 per 10,000 admissions (range 8.99-20.35). Most patients had a previously identified risk profile of old age, comorbidity and recent use of antibiotics. 41/276 (14.86%) patients had died, and 11/41 (26.82%) were CDI-associated deaths. Complicated CDI were registered in 53/276 (19.21%) patients, and recurrent infections in 65/276 (23.55%). Conclusion: Our data suggest that CDI is largely present in our setting which represents a serious problem and points to the importance of international surveillance, detection and control of CDI.

Objectives: Hospital outbreaks of multidrug-resistant Klebsiella spp., especially those in neonatal wards, are often caused by new types of strains, the so-called extended-spectrum-beta-lactamase (ESBL) producers. Methods: The aim of this study was to determine the distribution of nosocomial infections caused by Klebsiella pneumoniae ESBL strain by location and kind of infections in the University Clinical Centre Tuzla during a period of one year. A prospective study was implemented for all patients who developed hospital infections caused by Klebsiella pneumoniae ESBL strain during the period from 1st of January to 31st of December 2010 year. Determination of nosocomial infections was performed using standardized CDC criteria. Mandatory registration of hospital infections was done via a written application form which is used for surveillance. Results: Hospital infections caused by Klebsiella pneumoniae ESBL strain were reported 141 times, during the year 2010. Most common hospital infection were urinary tract infections 52 times (36,9%), followed by respiratory tract infections 43 times (30,5%), infection of the gastrointestinal system 20 times (14,2%), infections of surgical sites 12 times (8,5%), 10 times (7,1%) the bloodstream infection and other infections 4 times (2,8%). Nosocomial infection was mostly reported in the Clinic of anesthesiology and reanimation 37 times (26,2%) and Clinic for children\'s diseases 33 times (23,4%). Regarding age, 41 (29,1%) of nosocomial infections were detected in patients below the age of one year and 35 (24,8%) in patients above the age of 65 years. Conclusion: Respiratory tract infections were reported more frequently in the intensive care unit than in the wards. However, infection of the urinary tract and gastrointestinal infections were reported more frequently in the wards than in intensive care units.

ABSTRACT Glycoprotein L (gL), which complexes with gH, is a conserved herpesvirus protein that is essential for Epstein-Barr virus (EBV) entry into host cells. The gH/gL complex has a conserved role in entry among herpesviruses, yet the mechanism is not clear. To gain a better understanding of the role of gL in EBV-mediated fusion, chimeric proteins were made using rhesus lymphocryptovirus (Rh-LCV) gL (Rh gL), which shares a high sequence homology with EBV gL but does not complement EBV gL in mediating fusion with B cells. A reduction in fusion activity was observed with chimeric gL proteins that contained the amino terminus of Rh gL, although they retained their ability to process and transport gH/gL to the cell surface. Amino acids not conserved within this region in EBV gL when compared to Rh gL were further analyzed, with the results mapping residues 54 and 94 as being functionally important for EBV-mediated fusion. All chimeras and mutants displayed levels of cell surface expression similar to that of wild-type gL and interacted with gH and gp42. Our data also suggest that the role of gL involves the activation or recruitment of gB with the gH/gL complex, as we found that reduced fusion of Rh gL, EBV/Rh-LCV chimeras, and gL point mutants could be restored by replacing EBV gB with Rh gB. These observations demonstrate a distinction between the role of gL in the processing and trafficking of gH to the cell surface and a posttrafficking role in cell-cell fusion.

Abstract Alteration minerals dickite and kaolinite were detected in two hydrothermal Pb-Zn-Ag sulphide deposits in the northern Kosovo region. Dickite is associated with skarn mineralization in the Trepča (Stari Trg) deposit and kaolinite occurs in vein sulphide parageneses in the Crnac deposit. The mineralogical characteristics of dickite and kaolinite were determined by X-ray powder diffraction, Raman spectroscopy and scanning electron microscopy with EDS detector. Fluid-inclusion microthermometry and ion chromatography of leachates provided information on the P-T-X conditions of the genesis of dickite at Trepča. It was formed at temperatures between 290 and 330ºC and pressures between 12 and 60 MPa from a fluid with salinity in the range 6-8.5 wt.% NaCl eq. and pH <5.5. Kaolinite was deposited from a fluid with minimum temperatures between 210 and 250ºC, minimum pressure of 1.7 to 3.7 MPa, and salinity between 4.6 and 5.1 wt.% NaCl eq. Both dickite and kaolinite are related to the acidic pre-mineralization phase in the deposits.