Characterization and clonal representation of MRSA strains in Tuzla Canton, Bosnia and Herzegovina, from 2009 to 2017.

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.

Ocular Dirofilariasis in Bosnia and Herzegovina: A Case Report and Review of the Literature.

PURPOSE: To report a rare occurrence of ocular dirofilariasis in Bosnia and Herzegovina and provide a short overview on clinical characteristics and treatment options for ocular dirofilariasis. METHODS: A 75-year-old woman was admitted to the University Clinical Centre Tuzla with pain and redness in her left eye. A live, white, coiled, and active worm was noticed in subconjunctival space of bulbar conjunctiva. RESULTS: After successful surgical extraction, the parasite was identified as the adult form of Dirofilaria repens. There were no signs of intraocular or systemic inflammation. CONCLUSION: Ophthalmologists should have in mind a possible infestation, especially in cases with repeated inflammatory reactions and swelling of the ocular region, which does not respond to conventional therapy.

Analysis of Clinical and Laboratory Characteristics of Patients with Leptospirosis in Five-year Period.

INTRODUCTION: Leptospirosis is an infection caused by spiral bacteria from the family Leptospiraceae, and is considered to be the most widespread zoonosis in the world. AIM: To analyze the clinical and laboratory characteristics of patients with Leptospirosis over five years. METHODS: The study included 160 patients aged 17-79 years, who in the period 01.01.2014. to 21.12.2018. were hospitalized at the Clinic for Infectious Diseases of the University Clinical Center Tuzla. They were diagnosed based on medical history, clinical examination, laboratory and microbiological results. The definitive diagnosis was confirmed by serological testing from the patients' blood. RESULTS: In the observed period, the highest number of patients were in 2014 80/160, and the lowest in 2015 15/160. Male patients were more likely to suffer from 118 (73.8%) than female 42 (26,3%). The mean age was ±56.5 years. The most common symptoms in patients were: fever (95,6%), headache (93,8%), malaise (87,5%) and myalgia (85,6%). In all patients, 160/160 (100%) accelerated erythrocyte sedimentation and elevated C-reactive protein was observed. The following findings were reported from white blood cell findings: leukocytosis in 81/160 (50,6%), neutrophilia in 103/160 (64,4%) and lymphopenia in 128/160 (80%) patients. Liver findings had the following values: elevated AST in 142/160 (88%) and ALT in 130/160 (81,3%) patients. Total bilirubin was elevated in 105/160 (65,6%) and direct in 107/160 (66,9%). Thrombocytopenia was in 142/160 (88%) patients. Urea was elevated in 103/160 (64,4%) and creatinine in 101/160 (61,3%) patients. CONCLUSION: it is very important that physicians in their day to day practice, especially in ambiguous febrile conditions, differentially diagnose leptospirosis and establish a timely diagnosis, this is ley to adequate and timely therapy, and therefore to reducing to development of complications and mortality.

Presence and resistance of Streptococcus agalactiae in vaginal specimens of pregnant and adult non-pregnant women and association with other aerobic bacteria.

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.

THE FREQUENCY OF COMMUNITY-ACQUIRED METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (CA-MRSA) AMONG SAMPLES IN INSTITUTE FOR PUBLIC HEALTH IN CANTON SARAJEVO.

BACKGROUND: The increase in the incidence of methicillin-resistant Staphylococcus aureus (MRSA) infections lacking risk factors for exposure to the health care system has been associated with the recognition of new MRSA clones known as community-associated MRSA (CA-MRSA). These strains have been distinguished from health care-associated MRSA (HA-MRSA) strains by epidemiological, molecular and genetic means as well as by antibiotic susceptibility profile, tissue tropism and virulence traits. OBJECTIVE: To assess prevalence and antibiotic susceptibility profile of CA-MRSA in Canton Sarajevo, Bosnia and Herzegovina. RESULTS: Out of 1.905 positive Staphylococcus aureus isolates from various samples of outpatients collected during six months, 279 (14,64%) were MRSA isolates. Out of 279 MRSA samples, 133 (47,67%) were found in nasal swabs, from which 48 (36,09%) were in the age group <1 year and 39 (29,32 %) are in the age 1-5 year. Rate of the positive skin swabs was highest among the subject of age group <1 year (46 or 54,12 %) and 1-5 year (18 or 21,18 %). Predominantly antibiotic types among MRSA strains are resistant to penicillin and cefoxitin (36,90 %) and to penicillin, cefoxitin and erythromycin (61,35 %). CONCLUSION: Continued monitoring of epidemiology and emerging drug resistance data is critical for the effective management of these infections.

Diagnostic Significance of Reduced IgA in Children.

INTRODUCTION: The finding of reduced value of immunoglobulin A (IgA) in children is frequent in daily medical practice. It is important to correctly interpret the findings as adequate further diagnostic evaluation of the patient in order to make the determination on the significance of such findings. In children younger than 4 years always consider the transient impairment of immunoglobulins, maturation of child and his immune system can lead to an improvement in the clinical picture. In older children decreased IgA may lead to serious illnesses that need to be recognize and acknowledge through the appropriate diagnostic methods. At the University Clinical Center Tuzla, children with suspected deficient immune response due to reduced values of IgA, goes through further diagnostic evaluation at the Polyclinic for Laboratory Medicine, Department of Immunology and Department of Microbiology, as well as the Clinic of Radiology. MATERIAL AND METHODS: Our study followed 91 patients, for the year 2013, through their medical charts and made evaluation of diagnostic and screening tests. CONCLUSION: The significance of this paper is to draw attention to the importance of diagnostic approach to IgA deficient pediatric patient and relevance of knowledge of individual diagnostic methods as well as to the proper interpretation of the results thereof.

Diagnostic significance of reduced IgA in children.

INTRODUCTION: The finding of reduced value of immunoglobulin A (IgA) in children is frequent in daily medical practice. It is important to correctly interpret the findings as adequate further diagnostic evaluation of the patient in order to make the determination on the significance of such findings. In children younger than 4 years always consider the transient impairment of immunoglobulins, maturation of child and his immune system can lead to an improvement in the clinical picture. In older children decreased IgA may lead to serious illnesses that need to be recognize and acknowledge through the appropriate diagnostic methods. MATERIAL AND METHODS: Research was realized at the University Clinical Center Tuzla. Children with suspected deficient immune response due to reduced values of IgA observed and, goes through further diagnostic evaluation at the Polyclinic for Laboratory Medicine, Department of Immunology and Department of Microbiology, as well as the Clinic of Radiology. In the period of year 2013, there were a total of 91 patients with reduced values of IgA, age up to 13 years, of which 55 boys and 36 girls. RESULTS: Our study followed 91 patients, for the year 2013, through their medical charts and made evaluation of diagnostic and screening tests. The significance of this paper is to draw attention to the importance of diagnostic approach to IgA deficient pediatric patient and relevance of knowledge of individual diagnostic methods as well as to the proper interpretation of the results thereof.

Predictors of the therapeutic response of patients with influenza A (H1N1) 2009 infection.

AIM: To determine a predictive potential of biochemical and clinical parameters, including independent predictors of the therapeutic outcome in patients with H1N1 influenza. METHODS: In this retrospective study, 119 patients treated at the Clinic for Infectious Diseases in Tuzla in the period July 2009 to February 2010 were included. They were at the age of 1 month up to 80 years. The patients showed clinical, epidemiological and biochemical parameters of the pandemic flu and they had also been positive for the virus A (H1N1), which was confirmed by the real time polymerase chain reaction (RT-PCR). RESULTS: One hundred and nineteen patients were positive to the virus A (H1N1), 60 (50.4%) were male, and 59 (49.6%) female patients. The average age of the patients was 26.74 years. The number of patients with co-morbidity was 53 (44.5%). There were 71 (59.7%) patients who had been admitted to the Intensive Care Unit. The complications of the flu had appeared in 45 (37.8%) patients. Pneumonia was recorded in 108 (90.8%), and reduced saturation in 62 (52.1%) patients. The highest number of cured patients was in the group of patients who had received the oseltamivir within 48 hours after the outbreak of the disease. CONCLUSION: The age, reduced saturation with oxygen, duration of the hospitalization, pregnancy and overweight were independent predictors of the severe form of the disease/lethal outcome. For an optimal therapeutic outcome, it is important to apply antiviral therapy as soon as possible.

Comparison of double disk synergy test, VITEK 2 and Check-MDR CT102 for detection of ESBL producing isolates.

OBJECTIVE: This study is to define the statistical significance for detection of ESBL producers by the double disk synergy test and molecular test (Check-MDR CT102), microdilution test (VITEK 2 with AES) and double disk synergy test (DDST), as well as the microdilution test and molecular test. MATERIALS AND METHODS: Phenotypic testing of 55 isolates Enterobacteriaceae (Escherichia coli (14/55), Klebsiella pneumoniae (34/55), Klebsiella oxytoca (3/55) and Proteus mirabilis (4/55) was performed by VITEK 2 Compact/AES. When this test showed positive results for the ESBL phenotype, then DDST with amoxicillin/clavulanate, ceftazidime, cefpodoxime, aztreonam, ceftriaxone and cefoxitin disks was performed along with Check-MDR CT102 which identified CTX-M, TEM and SHV ß-lactamases. RESULTS: Applying the McNemar test, we determined that there was a statistically significant difference in the results of detection of ESBLs bacteria using DDST compared to molecular methods (95% CI=41.92 to 54.55; p<0.0001), as well as a DDST and VITEK 2/AES (95% CI=40.13 to 52.73; p<0.0001). We did not find any statistically significant difference in the results of detection of ESBL producers using molecular techniques and VITEK 2/AES (CI=-4,43 to 5,36; p=1). Also we did not find any statistical.. difference between the resistance to cefpodoxime and ceftriaxone (50/50) compared to the results of molecular tests. CONCLUSION: In routine daily testing, good detection of ESBLs bacteria, especially CTX-M can be obtained with phenotypic methods with VITEK 2/AES and by DDST with cefpodoxime, and ceftriaksone disks.

Etiology of bacterial meningitis in children in Tuzla Canton.

INTRODUCTION: Bacterial meningitis is one of the most serious infectious diseases in childhood. AIM: To identify the most common causes of bacterial meningitis in children in Tuzla Canton. PATIENTS AND METHODS: This is a retrospective study which included 140 children, aged from 1 month to 14 years, treated at the Clinic for Infectious Diseases Tuzla, in the period 1999-2009, who had clinical and laboratory parameters of bacterial meningitis. RESULTS: In Tuzla Canton in ten year period were registered 140 cases of bacterial meningitis in children aged 1 month to 14 years. Prevalence was 1.53/1000. The average age of patients was 3.6 +/- 3.8 years. Male to female ratio was 1.3:1. April, May and November were the months with the most reported cases of bacterial meningitis (p = 0.02). Most affected were children 1-12 months of age (p < 0.001). The most common pathogens of bacterial meningitis were Haemophilus influenzae (13.6%), Neisseria meningitides (8.6%) and Streptococcus pneumoniae (5.7%) (p < 0.001). Mortality was 2.14%. CONCLUSION: Bacterial meningitis is present in Tuzla Canton, and three major pathogens are Haemophilus influenzae, Neisseria meningitis and Streptococcus pneumoniae. The most affected were children 1-12 months of age, and most cases of bacterial meningitis occurred in spring.

The importance of serological tests implementation in disseminated candidiasis diagnose.

Candidiasis is defined as an infection or disease caused by a fungus of the genus Candida. Rate of disseminated candidiasis increases with the growth of the number of immunocompromised patients. In the the last few decades the incidence of disseminated candidiasis is in growth as well as the mortality rate. The aim of this survey is to show the importance of serological tests implementation in disseminated candidiasis diagnose. This is a prospective study involving 60 patients with malign diseases with and without clinical signs of disseminated candidiasis and 30 healthy people who represent the control group. Apart from hemoculture, detection of circulating mannan antigen and adequate antibodies of Candida species applying comercial ELISA test was determined in each patient. This survey deals with relevant factors causing disseminated candidiasis. This survey showed that the group of patients with clinical signs of disseminated candidiasis had more patients with positive hemoculture to Candida species, then the group of patients without clinical signs of disseminated candidiasis. The number of patients being examined and positive to antigens and antibodies was higher (p < 0.01) in the group of patients with clinical signs of disseminated candidiasis (7/30; 23.3%), then in the group of patients without clinical signs of disseminated candidiasis (0/30; 0%): Average value of titra antigen was statistically higher (p < 0.001) in patients with Candida spp. positive hemocultures rather then in patients with Candida spp. negative hemocultures. In the group of patients with clinical signs of disseminated candidiasis 6/30 (20%) of patients had Candida spp.positive hemocultures while in the group of patients without clinical signs of disseminated candidiasis 1/30 (3.3%) of patients had Candida spp. positive hemocultures, which was considerably higher (p < 0.05). Correlation of results of hemoculture and mannan antigens and antibodies in patients with disseminated candidiasis were statistically significant, while correlation of results of hemoculture and antibodies was insignificant. Because of low sensitivity of hemoculture and time needed for isolation of Candida spp., introducing serological tests in regular procedures would speed disseminated candidiasis diagnose.

Etiology of peritonitis.

AIM: To determine the etiology of peritonitis, as well as the sensitivity of isolated bacteria to antibiotics. METHODS: Study was performed at the Surgical Clinic, Department of Anesthesiology and Reanimation, Department of Microbiology, Department of Clinical Pharmacology of University Clinical Centre (UCC) Tuzla. In a prospective study, starting from 1.6.2009. to 30.6.2010., was 60 patients analyzed who underwent surgery for acute peritonitis in the Surgical Clinic, UCC Tuzla. RESULTS: After microbiological processing of the content of the operative field, wound swabs and smears of drainage, we isolated Gram positive coccid bacteria: Coagulase-negative Staphylococcus in 18.36%, Staphylococcus aureus in 12.6%, while Enterococcus faecalis and Enterococcus faecum were found in 3.44% isolates. In the group of Gram-negative bacteria we isolated: Escherichia coli in 15.51%, and Pseudomonas aeruginosa and Klebsiella pneumoniae in 10.34% of the cases. The sensitivity of coagulase-negative Staphylococcus was largest to vancomycin (100%), and lowest to penicillin (10%) and Staphylococcus aureus sensitivity was largest to vancomycin and amoxicillin-clavulanate (100%) and lowest to penicillins (3.7%). Sensitivity of the facultative anaerobic Gram-negative bacilli was highest to carbapenems 95-100%, and lowest to penicillin preparations 0-56%. CONCLUSION: Based on the results of microbiological findings of 83.33% patients empirical antibiotic therapy was appropriately involved, and in 16.67% was inadequate and later changed, following the results of microbiological findings. This study indicates the importance and validity of microbiological diagnostics to determine the etiology of peritonitis and appropriate selection of antibiotics to treat these infections, with the required surgical treatment.

Surveillance of wildlife zoonotic diseases in the Balkans Region.

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.

Importance of isolation and biotypization of Gardnerella vaginalis in diagnosis of bacterial vaginosis.

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.

Bacterial vaginosis presence in sexually active women in Tuzla Canton area.

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.

[Diagnostical values of antibody on A60 antigen in diagnostic of tuberculosis]. / Dijagnosticka vrijednost nalaza antitijela na A60 antigen u dijagnostici tuberkuloze.

Aim of this research was to investigate diagnostic value of discovering of antibody on A60 antigen in patients who were tested for presents of Mycobacterium tuberculosis in there biological samples. We tested a samples of sputum, gastric juice, urine, cerebrospinal fluid and punctate from group of 353 patients who were suspected for tuberculosis. In all patients we were looking for antibodies classes A60 antigen. We used immune chromatographic "Hexagon TB" test, Germane company "Human Geselschaft fur Biochemica und Diagnostica". From 353 patients we found 58 (16.43%) patients with positive BK, 79/22,38%) patients with positive Lowenstein culture and 122 (34,55%) patients with antibody in sera on A60 antigen. Patients who were BK and Lowenstein positive, have had antibody in 94,23% cases, Patients who were BK negative and Lowenstein positive have had antibodies in 70,37% cases and patients who were BK negative and Lowenstein negative have had antibody in 19,03% cases. Patients with BK positive and Lowenstein negative results have had antibody in 50,00% cases. Difference between results is significant (p<0,01). From 122 patients with positive antibodies, 52 were BK positive and 68 have had positive Lowenstein cultures. From 231 patients with no antibody, just 6 were BK positive and 11 Lowenstein positive. In 62 patients with positive antibodies, were BK and Lowenstein negative. We confirmed that antibody on A60 antigen in microbiological positive patients are more often then in microbiological negative patients (p<0,001).

[Comparison of different methods for the detection of enteroviruses in drinking water]. / Usporedba razlicitih metoda koncentriranja enterovirusa iz vode te detekcija virusnog genoma (metodom reverzne transkripcije i lancanom reakcijom polimeraze).

UNLABELLED: Detection of enteroviruses in drinking water poses a very specific problem, since a very small number of particles have to be identified in huge water quantities. Currently, there are a number of methods to identify the concentration of virus particles and RNA templates to carry out RT-PCR, however, no standard method has yet been proposed. AIM: The aim of this report is to suggest optimal methods for the preparation of RNA templates to carry out RT-PCR. MATERIAL AND METHODS: In this experimental study, two different methods were employed on preparing RNA template. The concentration of virus particles in a large (10 L) and small (1 L) quantity of water was determined by use of the electropositive microporous virology filter (method 1 and method 2). Elution and flocculation of the virals particles were performed by organic extraction (method 1) and inorganic extraction (method 2). The sensitivity of the methods was assessed by testing the artificially contaminated water with 10(1) to 10(5) virus-particles using 1-L and 10-L containers of water. RESULTS: Method 1 detected 10(5) and 10(4) virus particles, method 2 10(5), 10(4) and 10(3) virus particles from 10 L and 1 L of water, respectively, yielding a statistically significant difference (p<0.01; chi2 = 6.061). Using two-step RT-PCR with nested PCR method, enteroviruses were detected in 42/100 (42%) samples of surface drinking water and in 83% of the same samples using RT-PCR without nested PCR.

[Prevalence of patients infected with resistant strain of Mycobacterium tuberculosis in Canton Tuzla]. / Prevalencija bolesnika inficiranih rezistentnim sojevima Mycobacterium tuberculosis na Tuzlanskom kantonu.

Aim of this work is to show the level of prevalence of patients infected with resistant strains Mycobacterium tuberculosis in Canton Tuzla. In the period of 1996-2003 year we tested 87,408 samples of different materials on existence of Mycobacterium tuberculosis. Among all samples there were 66,128 sputum, 14,599 urines, 3,817 gastric juice, 1,174 materials from broncholavage and 547 other samples. Microscopically it was found 4,380 smear-positive samples and 6,365 samples were positive on Loewenstein medium. Positive sputum had 1,917 patients, and positive culture had 3,018 patients. Resistance test was done on streptomycin, isoniazid, rifampicin and ethambutol with standard proportional method for 2,662 patients. Totally sensibile were 2,570 or 96.54%, and restant were 92 or 3.46% patients. Patients infected with mono-drug resistant strains Mycobacterium tuberculosis were 71 or 2.67%, and poli-drugs resitant 21 or 0.78%. There were 16 patients or 0.60% infected with multi-drugs resistant strains. Time of bacteriological negativization for the patients infected with resistant strains was in the average 8.19 months, for the patients infected with mono-drug resistant strains was 2.75 months and for infected with multi-drugs resistant strains was 32 months. It is concluded that region of Canton Tuzla has high level of bacteriological prevalence but low level of prevalence of patients infected with resistant strains Mycobacterium tuberculosis in the this period of time, and it is significantly lower then earlier periods, thanks to national tuberculosis control program and system of directly observed treatment.

[Viruses in drinking water: HAV and enteroviruses]. / Virusi u vodi za pice: HAV i enterovirusi.

The presence of waterborne enteric pathogens (bacteria, viruses and protozoa) in potable water represents a potential danger for people's health. While doing a microbiological examination of potable water on a regular basis, we obtain a constant control of presence of the bacteria that contaminate water. However, the water examination of the enteric viruses is done only when we have some large-scale hydrous epidemics. Finding the bacteria wich point to fecal contamination of drinking water is not a safe indicator of presence of the enteric viruses. After a two step RT/PCR examination on viruses in 84 samples of potable water, the enteroviruses were found in 53 out of 84 samples (63.09%), while the hepatitis A virus was not found after one step RT/PCR examination. In conclusion, we emphasize the need of standardizing the method of potable water viruses detection, in order to legalize the need of its regular application.