The countries of the Balkan Peninsula have become the region with frequent outbreaks of the emerging and re-emerging diseases during the last decade of the 20th and the first decade of the 21st century. The majority of outbreaks were wildlife zoonotic, and vector-borne diseases, such as brucellosis, leptospirosis, listeriosis, tularemia, Q-fever, Lyme disease, anthrax, rabies, viral hemorrhagic fevers, sandfly fever, tick-borne encephalitis and leishmainiasis. Epidemiological factors determined by ecology of causative agents are often the most useful diagnostic clues. The recognition of evolving problems of emerging and re-emerging diseases emphasizes the need for the development of better laboratory diagnostic methods for the surveillance and tracking of the diseases, and for continued research of factors contributing to the transmission of the organisms. The continuous occurrence of previously unidentified infections requires prospective national strategies for timely recognition of the syndromes, causative agent identification, establishment of criteria and methods for the diagnosis, optimization of the treatment regime, and determination of successful approaches to prevention and control. Wildlife diseases surveillance in the most of the Balkan countries has been coordinated by the WHO since 1992. Although new technology and communication have extremely improved in the last decade, there is a need for optimal communication lines among the Balkan countries, better exploitation of communication technologies like the Internet and other media in the field of emerging diseases.

Aim:  The clinical course and outcome of patients with haemorrhagic fever with renal syndrome (HFRS) caused by Puumala (PUUV) and Dobrava viruses (DOBV) were analyzed and whether it left long‐term consequences on kidney function after 10 years was evaluated.

Postoperative wound infections represent about 16% of hospital-acquired infections. Staphylococcus aureus is the most common cause of nosocomial wound infections. Increased frequency of Methicillin-resistant Staphylococcus aureus (MRSA) in hospitalized patients and possibility of vancomycin resistance requires permanent control of MRSA spread in the hospital.The purpose of this study was to analyse the frequency of Methicillin-resistant Staphylococcus aureus (MRSA) in the swabs taken from the surgical wounds, the presence of MRSA infection in surgical departments and to examine antimicrobial susceptibility of MRSA isolates. Wound swabs were examined from January 2006 to December 2008. The isolates were identified by conventional methods. Antimicrobial susceptibility testing was performed by Kirby-Bauer disc-diffusion method as per NCCLS guidelines.A total of 5755 wound swabs were examined: 938 (16,3%) swabs were sterile and 4817 (83,7%) were positive. Staphylococcus aureus was isolated in 1050 (22,0%) swabs and it was the most common cause of wound infections. MRSA was isolated from 12,4% samples in 2006, from 6,7% samples in 2007 and from 3,7% samples during 2008. Wound infections caused by MRSA dominated in the department of plastic surgery (24,4%) and in the department of orthopaedic surgery (24,1%). Antimicrobial susceptibility testing showed that 73% of MRSA isolates were with the same antibiotic sensitivity pattern (antibiotyp)-sensitive only to vancomycin, tetracycline, fucid acid and trimethoprim/sulfamethoxasole. Our results show decreasing of MRSA infection in the surgical wards. These results appear to be maintained with strategies for preventing nosocomial infection: permanent education, strong application of protocols and urging the implementation of strict infection control policy.

Pregnancy represents a risk factor in the occurrence of vaginal candidosis. The objectives of our study were: to make determination of the microscopic findings of vaginal swab, frequency of Candida species in the culture of pregnant women and patients who are not pregnant, determine the Candida species in all cultures, and to determine the frequency and differences in the frequency of C. albicans and other non-albicans species. In one year study performed during 2006 year, we tested patients of Gynaecology and Obstetrics clinic of the Clinical Centre in Sarajevo and Gynaecology department of the General hospital in Sarajevo. 447 woman included in the study were separated in two groups: 203 pregnant (in the last trimester of pregnancy), and 244 non-pregnant woman in period of fertility. Each vaginal swab was examined microscopically. The yeast, number of colonies, and the species of Candida were determined on Sabouraud dextrose agar with presence of antibiotics. For determination of Candida species, we used germ tube test for detection of C. albicans, and cultivation on the selective medium and assimilation tests for detection of non-albicans species. The results indicated positive microscopic findings in the test group (40,9%), as well as greater number of positive cultures (46,8%). The most commonly detected species for both groups was C. albicans ( test group 40.9% and control group 23,0%). The most commonly detected non-albicans species for the test group were C. glabrata (4,2 %) and C. krusei (3,2%), and for the control group were C. glabrata (3,2%) and C. parapsilosis (3,2%). The microscopic findings correlated with the number of colonies in positive cultures. In the test group, we found an increased number of yeasts (64,3%), and the pseudopyphae and blastopores by microscopic examination as an indication of infection. In the control group, we found a small number of yeasts (64,6%) , in the form of blastopores, as an indication of the candida colonisation. Our results indicate that gravidity, as the risc factor for incidence of infection, has the significant role in the incidence of vaginal candidosis.

Fifty-seven nosocomial Klebsiella pneumoniae isolates producing extended-spectrum beta-lactamases (ESBLs) were collected between February 2007 and November 2007 in different wards of the Sarajevo (Bosnia-Herzegovina) reference hospital. These isolates comprise two major epidemic pulsed-field electrophoresis-defined clones plus two minor clones. In addition to the ESBL-mediated resistance, all strains uniformly showed resistance to ciprofloxacin, gentamicin and tobramycin. The beta-lactamases involved in this resistance phenotype were TEM-1, SHV-1, and CTX-M-15, as demonstrated by isoelectric focusing, PCR amplification, and sequencing. TEM-1 and CTX-M-15 beta-lactamases, as well as the aminoglycoside resistance determinants, were encoded in plasmids that could be transferred to Escherichia coli by conjugation. In three of the infected patients with the predominant clone, cefoxitin resistance development (MICs >128 mg/L) was documented. The analysis of the outer membrane proteins of the cefoxitin-susceptible and cefoxitin-resistant isolates revealed that the former expressed only one of the two major porins, OmpK36, whereas in the latter, the expression of Ompk36 was altered or abolished. This is the first report of CTX-M-15-producing K. pneumoniae in Bosnia-Herzegovina. Furthermore, we document and characterize for the first time cefoxitin resistance development in CTX-M-15-producing K. pneumoniae.

UNLABELLED Brucellosis is a worldwide zoonosis with a high degree of morbidity in humans. In Bosnia and Herzegovina a progressive increase of brucellosis among humans is evident. As the clinical picture of human brucellosis is fairly non-specific, a definitive diagnosis requires isolation of the causative organism, or the demonstration of the high levels of specific antibodies, or seroconversion. AIM To analyse the diagnostic value of the Rose Bengal test, blood culture and immunoenzymatic test (ELISA IgM and IgG) in patients with brucellosis and to examine the relationship between these diagnostic methods. METHODS We analysed the diagnostic methods in 525 brucellosis patients from 2004 to 2008. All patients were treated at the Infectious Diseases Clinic, University of Sarajevo Clinics Centre. The disease was diagnosed by positive blood culture results and/or by positive relevant serologic test results (ELISA, Rose-Bengal plate-agglutination test). RESULTS In total 162/525 (30.8%) patients had positive blood cultures. The Rose Bengal test was positive in all patients--525/525 (100.0%). Brucella IgM antibodies with ELISA were positive in 341/525 (64.8%). Early in infection, antibodies of the IgM class predominate. Brucella IgG antibodies with ELISA were positive in 236/525 (56%). CONCLUSION This study clearly showed that only a combination of blood culture, Rose Bengal test and ELISA ensured early and precise diagnosis of human brucellosis. The Rose Bengal test is excellent for screening. Blood culture gave excellent results in patients with primary infections. ELISA(IgM, IgG) is the method of choice for the diagnosis of chronic disease and relapse.

SUMMARY The extent of hantavirus seroprevalence in the healthy population from Bosnia and Herzegovina has not yet been investigated; therefore, the aim of this study was to assess the hantavirus seroprevalence in the population from different regions of Bosnia and Herzegovina and in different risk groups. The serosurvey included 1331 subjects from endemic and non-endemic regions in Bosnia and Herzegovina. All sera samples were examined using IgG ELISA, and Western blot (Bunyavirus IgG) tests. Hantavirus seroprevalence was 7·4% in the endemic region and 2·4% in the non-endemic region (P<0·05). Former soldiers from the endemic region had significantly the highest seroprevalence (16·1%) compared to the general population from the endemic region (6·2%), the occupational risk group from the non-endemic region (5·6%) and the general population from the non-endemic region (0·8%) (P<0·01). No difference in hantavirus seroprevalence between gender or age groups was observed. Hantavirus seroprevalence in different populations in Bosnia and Herzegovina was found to be highest compared to other central European countries.

Brucellosis is an important public health problem in Bosnia and Herzegovina. The diagnosis of brucellosis in the country without any experiences with this kind of infection may be very difficult. The aim of this study was to evaluate diagnostic methods: Rose Bengal test, blood cultures and ELISA IgM and IgG in the patients with brucellosis. The study included 91 brucellosis patients in the period 2004 to 2007. All the patients were treated at the Clinic for Infectious Diseases, University of Sarajevo Clinics Centre. Blood cultures were positive in 28/91 (30, 8%) patients. This method often needs a long period of incubation and specimens need to be obtained early. These limitations make serology the most useful tool for the laboratory diagnosis of Brucella infection. Rose Bengal is a rapid plate agglutination test, very sensitive irrespective of the stage of the disease. In our study, Rose Bengal test was positive in all patients 91/91 (100, 0%). Brucella IgM antibodies with ELISA were positive in 59/91 (64, 8%). Brucella IgG antibodies with ELISA were positive in 51/91 (56%). In order to determine the diagnostic value of the different tests, we compared the sensitivity among test-methods: Rose Bengal test-100.0%, blood culture-30.8%, ELISA IgM-64.8% and ELISA IgG-56.1%. Sensitivity of test methods was different in the different stages of illness. It is necessary to use combination of different tests such are blood culture, Rose Bengal test and ELISA in order to ensure the diagnosis. Rose Bengal test is excellent for the screening. Blood culture is a method of choice for the diagnosis acute infection. ELISA is a very good method for the diagnostic chronic disease and relapse.

Diarrhoeal disease is a major cause of illness and death among infants and young children worldwide. Among the Escherichia coli (E. coli) causing intestinal diseases, there are six well-described categories: enteroaggregative E. coli (EAEC), diffusely adherent E. coli (DAEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enterohaemorrhagic E. coli (EHEC) and enterotoxigenic E. coli (ETEC). The aim of the present study was to investigate the relative contribution of different groups of diarrhoeagenic E. coli (DEC) in paediatric patients with diarrhoea. Clinical stool specimens from 380 children with diarrhoea, with ages ranging from birth to < 12 years, were selected for the study over a period of 17 months (August 2007 to December 2008). The study showed that 85/380 children (22%) had diarrhoea due to diarrhoeagenic E. coli. The most prevalent was enteropathogenic E. coli (EPEC) isolated from 46/85 paediatric patients (54%), followed by enterotoxigenic (ETEC) isolated from 19/85 (22.3%), enterohaemorrhagic (EHEC) from 18/85 (21.1%) and enteroinvasive (EIEC) from 2/85 patients (2.3%). The most prevalent serotypes of EPEC were O86:K61 and O44:K74 isolated from 10/46 (21.7%), O128:K67 from 6/46 patients (13%), followed by O158:K- and O126:K71 isolated from 4/46 patients (8.6%). Among the ETEC the most prevalent serotypes were O78:K80 isolated from 10/19 (56.7%) and O25:K11 from 9/19 patients (47.3%), especially during the first twelve months: 9/19 patients (47.3%). The most prevalent EHEC strain found in this study was O145:K- and O103:K-: 5/18 patients (27.8%). Two isolated strains of EIEC belong to serotype O164:K-. The average age of the patients was 2 years. Two patients with bloody diarrhoea had EHEC serotype O157:H7 which progressed to haemolytic-uremic syndrome (HUS). Our study shows that diarrhoeagenic E. coli is a significant causal agent of diarrhoeal diseases in paediatric patients in Bosnia and Herzegovina. This study is the first report about the frequency and most common serotypes of DEC in Bosnia and Herzegovina. Additionally, it is the first report of cases with an O157:K- infection which progressed to HUS, a serious and potentially fatal illness.

Urinary tract infection is commoner in patients with spinal cord injuries because of incomplete bladder emptying and the use of catheters that can result in the introduction of bacteria into the bladder. 145 patients suffering from spinal cord injuries, admitted to the Institute for physical medicine and rehabilitation, Centre for paraplegia of the Clinical Centre of the University of Sarajevo, were included. The patients were divided in three groups according to the method of bladder drainage: Group A (n=61) consisted of patients on clean intermittent catheterization; Group B (n=54) consisted of patients with indwelling catheters; Group C (n=30) consisted of patients who had performed self-catheterization. From a total of 4539 urine samples, 3963 (87,3%) were positive and 576 (12,7%) were sterile. More than 90% of the infected patients were asymptomatic. The overall rate of urinary infection amounted to about 2,1 episodes, and bacteriuria to 8,1 episodes per patient. 77% of infections (113/145) were acquired within seven days from catheterization. Infection was usually polymicrobial; the greatest number of urine samples 1770/3943 (44,9%) included more than one bacterium. The vast majority of cases of urinary tract infection and bacteriuria are caused by Gram-negative bacilli and enterococci, commensal organisms of the bowel and perineum, representative of those from the hospital environment. Providencia stuarti (18,9%) being the most common, followed by Proteus mirabilis (16,3%), Escherichia coli (11,8%), Pseudomonas aeruginosa (10,2%), Klebsiella pneumoniae (8,1%), Morganella morgani (5,4%), Acinetobacter baumannii (4,6%), Providencia rettgeri (3,5%). 15,7% of isolates were Gram-positive with Enterococcus faecalis (8,6%) as the most common. 55,3% of isolates were multidrug-resistant, and the highest rates of resistance were found among Acinetobacter baumannii (87,8%), Providencia rettgeri (86,7%), Pseudomonas aeruginosa (85,4%), Providencia stuarti (84,3%) and Morganella morgani (81,0%). Lower rates of resistance were found in Group C, i.e. patients on intermittent self-catheterisation. Eradication of organisms was achieved in only 53 (10,05%) of patients; hence, antibiotic therapy had no or very low effect. Significant correlations were found between the method of catheterization and the frequency of bacteriuria and urinary tract infections. The analysis of Group C showed a rate of lower urinary tract infection and bacteriuria than the other two Groups of patients. The objective of this study is the update of etiology and antimicrobial susceptibility in urinary tract infections in this group of patients. In addition, possible correlations between UTI and the type of bladder management were examined.

Sandfly fever viruses (SFV) are endemic in the Mediterranean, Middle East, northern African and western Asian countries. Toscana virus (TOSV), serotype of Sandfly fever Naples virus, is among of the three most prevalent viruses associated with meningitis during the warm seasons in northern Mediterranean countries. The historical data of the sandfly fever (Pappataci fever) indicates its origin in Bosnia and Herzegovina at the end of 19th century. There is a long period of time for which there are no data on research related to the SFV in Bosnia and Herzegovina. The purpose of the study was to investigate the presence of sandfly fever in Bosnia and Herzegovina in recent years. The 68 of serum samples were obtained from February 2006 until September 2008 from a group of patients with febrile illness of unknown etiology. The sera were tested on the presence of IgG and IgM antibodies against TOSV by specific serology test- recomLine Bunyavirus IgG/IgM immuno-line assay. The recent TOSV-infection was confirmed in the patients in each year during the study: 10,71% (3/28) in 2008; 9,38% (3/32) in 2007 and 12,50% (1/8) in 2006. The presence of specific antibodies to TOSV in the sera of the patients in recent years indicates re-emerging character of the disease in this region. It would be necessary to make biological, epidemiological and clinical research on the TOSV and related phleboviruses to elucidate the problem of SFV in Bosnia and Herzegovina.

Emerging and re-emerging infectious diseases create constant and serious concerns for public health. The majority of emerging infectious diseases (EID) are wildlife zoonotic diseases and vector-borne diseases. Croatia has a long tradition in the control, management and research of EID zoonotic diseases and vector-borne diseases. There has also been a long and advantageous tradition in the collaboration of different experts and professionals in EID research in Croatia involving physician clinicians in infectious diseases, microbiologists, pathologists, veterinarians and animal scientists, ecologists, forestry experts, wildlife scientists, public health specialists and epidemiologists and laboratory scientists. The University Hospital for Infectious Diseases in Zagreb established the Centre for Emerging and Re-emerging Infectious Diseases in liaison with national and international partners from Europe and the United States. This Centre is working in line with the 'One Health initiative' which recognises the inter-relationships between human, animal and environmental health.

The goal of our research was to determine the presence of bacterial vaginosis in sexually active women in Tuzla Canton area. Diagnosis determination for bacterial vaginosis was conducted on the basis of three out of four internationally accepted criteria according to Amsel and isolation and identification of Gardnerella vaginalis (G. vaginalis) by standard microbiological procedures. Bacterial vaginosis was diagnosed in 20,5 % (41/200) women who asked for gynaecologist's help due to their personal discomfort, since significantly higher percentage of diagnosed bacterial vaginosis of 48,80% (41/84) was determined in women with personal discomfort typical for this disease. All relevant factors, according to available literature, for genesis of bacterial vaginosis were processed in this research. In respect to the obtained outputs, bacterial vaginosis is significantly more frequent occurrence in women who are not married, since the number of sexual partners, the time of the first sexual intercourse, the use of intrauterine contraceptive device and smoking do not cause the genesis of bacterial vaginosis. According to Nugent, an increased vaginal discharge with unpleasant odour after sexual discourse, its pH>4,5, a positive amino odour test, an occurrence of clue cells in a direct microscopic concoction of vaginal discharge and assessment of the state of vaginal flora for bacterial vaginosis are significantly more frequent occurrences in women with individual discomforts. It was proved that G. vaginalis is a dominant micro organism in 95% of women with clinical signs of vaginosis although it was isolated from vaginal discharge in 40 to 50% of healthy women. In our research, G. vaginalis was isolated in 63,41% of examined women with all signs of bacterial vaginosis, in 36,59% of examined women with one or more clinical signs of bacterial vaginosis and in 2,58% of examined women of control group without clinical signs.

The natural habitat of Gardnerella vaginalis is a vagina since it could be located among 69% of women who have no signs of vaginal infection and in the vagina of as many as 13.5% girls. G. vaginalis is almost certainly identified among women diagnosed with bacterial vaginosis as well as in the urethra of their sexual partner. The increase in prevalence and concentration of G. vaginalis among patients diagnosed with this syndrome confirms that G. vaginalis plays a significant role in its pathogenesis. In our research, based on Amsel criteria for three or more clinical signs of bacterial vaginosis, it was diagnosed in 20.5% of women with subjective problems of vaginal infection, and in 48.80% of women with subjective symptoms characteristic of this disease. G. vaginalis was isolated from vaginal secretion of women without clinical signs characteristic of bacterial vaginosis. In 2.58% of cases it was solitary, while in 1.28% it was found in combination with other aerobic and anaerobic bacteria and, in 1.28% women combined with Candida albicans. The isolation of G. vaginalis was significantly increased (p<0.05) in the group of women with clinical signs of bacterial vaginosis in comparison to the group of women without these signs. Frequent recurrence of bacterial vaginosis, which is found in 20-30% of women within a three months treatment, is explained as reinfection with other biotype of G. vaginalis, different from a source biotype or as a consequence of wrong treatment. Following Piot biotype scheme, biotypes 2., 3. and 7. G. vaginalis are significantly more often isolated from women who suffer from bacterial vaginosis. Biotype 7. G. vaginalis, isolated from the group of women without clinical signs of bacterial vaginosis, accounted for 2.58% cases. Following Benit biotype scheme, biotypes IVa, IVc and IIc were identified in 12.90% cases, while biotypes IIIa, IIa, Ia, IVb, IIb were found in 6.45% cases. Lipase-positive isolates of G. vaginalis were significantly more frequently accompanied by the syndrome of bacterial vaginosis.

Importation of viruses from other continents caused prolonged circulation and large outbreaks in the WHO European Region.