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H. Resić, A. Ćorić, A. Dedeić-Ljubović, M. Hukić, E. Avdić, N. Kukavica

INTRODUCTION The prevalence of MRSA (Methicillin Resistant Staphylococcus Aureus) in haemodialysis patients has increased dramatically during recent years. The aim of our study is to determine the prevalence of MRSA colonization among haemodialysis patients and medical staff. MATERIAL AND METHODS This prospective study included 235 patients undergoing haemodialysis therapy and 60 medical staff members, in the Center for Haemodialysis Sarajevo. Nasal and throat samples were taken (identification of MRSA was performed using standard microbiological methods). A total of 474 nasal and throat cultures from patients' samples and 120 cultures from medical staff samples were obtained. RESULTS AND DISCUSSION The total number of sampled patients was 235 and colonies were found in 36 of the samples (15.3%). Among medical staff nasal carriage rate of MRSA was 11.6% (7/60). The mean age of haemodialysis patients with MRSA was 52.94:1:5.3. The patients aged between 55 and 64 had the highest prevalence of MRSA (11/36, 30.55%). Those aged 45-54 had the next highest prevalence of nasal carriage (10/36, 27.77%). Patients aged 65 years had middle prevalence of MRSA (6/36, 8.33%). Patients aged 25-34 had the lowest prevalence of nasal carriage. We did not have possibilities to isolate MRSA positive dialysis patients, and we used intensive antibacterial prophylaxis. We treated our patients with mupirocin ointment (three times a day for 5-14 days) and gained decolonization in 34 patients (94.4%). In the treatment period, positive medical staff did not come to work. CONCLUSION The overall prevalence of MRSA colonization in our dialysis patients was higher (15.3%). Implementation of adequate strategies for prevention of MRSA with application of mupirocin among carriers, reduced prevalence of MRSA in our dialysis units.

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.

S. Heljić, M. Hukić, A. Džinović, M. Babić, M. Djozic

Background. Invasive candidiasis in neonates has become an increasing problem over the past decade in Neonatal Intensive Care Units (NICUs) in the world; it is a relatively common cause of late onset sepsis associated with a high mortality. Risk factors are preterm babies who require invasive diagnostic and aggressive therapeutic procedures, prior antibiotic therapy, congenital abnormalities, gastrointestinal tract diseases, necrotizing enterocolitis and anatomical abnormalities requiring surgery. Prior colonization is the major risk factor for candidemia.Methods. To determine the rate of colonization, risk factors and the possible modes of acquisition of Candida spp. In neonates in NICU, we conducted a prospective cohort study including 48 successive admissions at NICU of Pediatric University Hospital, Sarajevo, during 3 months. Samples (mucocutaneous swabs, urine, stool, blood) were obtained weekly until the time of discharge or death (one infant enrolled in the study exited). Care health worker hands were cultured weekly (55 samples).Results. Candida spp. from various body sites samples were isolated in 7 patients (rate of colonization 14,8%) while 7 samples of health workers hands were positive for C. spp (14,5%). Patients colonizated or infected by Candida spp. In our study were or very preterm newborns (3/7) or compromised term newborns (4/7), with congenital abnormalities (2/7) or gastrointestinal surgical interventions (2/7). Two very preterm neonates were colonized early from the birth, which indicates possible vertical transmission. In other 5 patients, colonization occurred more lately, after at least 15 days stay in NICU, which indicates nosocomial transmission as a predominant mode of acquisition. Among 10 positive patterns from different body sites, Candida albicans was isolated in 6 cases; other 4 cases were C. glabrata (2/10), C. krusei (1/10) and C. famata (1/10).Conclusions. Rather than focusing on methods of identifying of pregnant women with vaginal Candida colonization, attention should be directed to high risk neonates and infection control measures including hands washing by health care workers.

D. Tulumović, E. Mesic, M. Hukić, N. Zivkovic

Objectives  Bosnia and Herzegovina is a known endemic region for Hantavirus infection. In this region at least two different Hantaviruses, the Dobrava viruses (DOBV) and Puumala viruses (PUUV) have been found. The aim of the study was to determine glomerular filtration rate (GFR) in patients with a history of hemorrhagic fever with renal syndrome (HFRS) 9 to 10 years after the acute phase of the disease. Design and Methods  We prospectively reviewed clearance of technetium marked diethyl three amino pentacetic acid (Tc99m DTPA) in 42 male convalescents with serologically confirmed HFRS, 13 patients with PUUV infection and 29 patients with DOBV infection. Results  The tubulosecretory phase of the GFR was prolonged in six patients (46%) with PUUV infection and 23 patients (79%) with DOBV infection (P < 0.05). Total and separate clearance EDTA were significantly statistically lower (P < 0.001) in patients with serologically confirmed DOBV infection. In patients with DOBV infection elimination of radio pharmac was prolonged. Conclusions  Hemorrhagic fever with renal syndrome can cause sustained renal damage that very rarely progresses into renal failure. The tubulosecretory DTPA phase was prolonged in 46% of examinees with PUUV and in 79% of examinees with DOBV infection. Elimination of radio pharmac from the kidney was prolonged in both groups of patients. The DTPA clearance rate was lower in patients with serologically confirmed HFRS than in normal populations, especially in patients with DOBV infection.

The target organ for Puumala (PUU) and Dobrava (DOB) viruses is the kidney, but the impact on renal function seems far more important in DOB infection than in PUU infections. Aim of study was to estimate the renal failure and capillary leak during the acute stage of DOB-infection and PUU-infection. 50 patients with serological conformed diagnosis of acute hantavirus infection were included in the study. Diuresis, serum urea and creatinine were measured. Size of kidneys and presence of ascites was followed by ultrasonography. Enlargement of both kidneys were found in all (100%) patients. Ascites was present in 8/25 (32%) DOB patients; in 1/25 (4%) PUU patient and pleural effusion in four 4/25 (16%) DOB patients. The statistically significant difference in the mean urinary output was during 5th, 6th and 7th day of hospitalization (P<0,05). Serum creatinine and urea levels were at significantly higher levels for DOB than for PUU patients (P<0,05). Acute haemodialysis was needed in 7/25 (28 %) of the DOB patients. One DOB patient died. DOB virus infection is associated with a more severe kidney function disorders than PUU virus infection. Capillary leak, which caused swollen of kidney and ascites is probably the pathogenesis key factor.

J. Karamehić, M. Hukić, F. Gavrankapetanović, B. Heljić, E. Hodzic, Z. Delibegović, Vanja Karlović

UNLABELLED The paper analyzed the values and titer of IgM and IgG antibodies on CMV in donors and recipients of kidney transplntats. The objective was to investigate the presence of CMV in donors and recipients. SUBJECT AND METHODS Test enzignost anti CMV/IgM was used. Detection of antibodies in 15 donors before transplantation and in 22 patients-recipients before and after the transplantation was done. RESULTS From total of 15 donors, the values of IgM ranged from 0,002 I.U. do 0,080 I.U. One case was positive. Four patients were seropositive and treated by Gancyclovirom 10 mg/kg/b.w (body weight) in period of 3 month. IgM values ranged from 0.88399 to 25978. The control finding 3 patients was negative, in 1 patient positive (IgM 0.569). In 18 patients the finding was negative. Refererent value for IgM is < 0.10 the result is negative. For IgG four times greater value comparing to the basic value was found. CONCLUSION The most significant factor for reactivation of CMV infection is iatrogenic Immunosuppressive therapy. Exclusion of CMV infection in donors and negative finding in recipient decrease.

: 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.

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.

S. Biel, O. Donoso Mantke, K. Lemmer, A. Vaheri, Å. Lundkvist, P. Emmerich, M. Hukić, M. Niedrig

D. Tulumović, M. Hukić, E. Mesic, Sana Sabović, A. Tulumović

INTRODUCTION There are significant changes in small blood vessels with increased permeability, edema of abdominal organs and ascites in the acute phase of hemorrhagic fever with renal syndrome (HFRS). We analyzed ultrasound changes of abdominal organs in patients with serologically verified HFRS, in order to assess the value of this method in the diagnosis of acute renal failure (ARF). METHODS The study included 14 sporadic patients with ARF and suspicion of HFRS, treated in 2002. Ultrasonography of the liver, gallbladder, pancreas, spleen, kidneys, prostate and retroperitoneum was performed on a Toshiba 220 SSA. RESULTS Parenchymal edema with increased echogenicity, sinus narrowing and enlargement of both kidneys were found in all 14 (100%) patients. Hepatosplenomegaly was found in seven (50%), ascites in four (28.6%), and pleural effusion in two (14.3%) patients. Increased thickness of the bladder wall was found in two (14.3%), gallbladder enlargement in one (7%), and edema of the pancreas in one (7%) patient. CONCLUSION Ultrasound examination has a very important role in the diagnosis and follow-up of changes in the parenchymal organs in the acute acute phase of the disease.

UNLABELLED Bosnia and Herzegovina has been known as a highly endemic region for Hantavirus infections for more than 50 years. Previous studies have shown that at least two different hantaviruses, the murine Dobrava (DOB) and avricoline Puumala (PUU) viruses, each carried by a different rodent species, have been circulating in the area. However, there is little information on rodent population density fluctuations in Bosnia over the past years as well as on the ratio of Puumala to Dobrava infection in humans. THE AIMS THE AIMS OF OUR STUDY WERE to identify the rodent species which may serve as hantavirus reservoirs in the north-east and central Bosnia; to assess the geographical distribution, density and population dynamics of rodent species in the area; to assess the influence of climatic conditions on the size of rodent population; and to determine the ratio of Puumala to Dobrava infection in humans. METHODS The epidemiologic and epizootic study in the north-east and central Bosnia was conducted during the 8-year period (1995-2003). The average yearly and monthly temperatures, air humidity and precipitation during the study period were analyzed. A total of 381 small rodents were caught during the epidemic years (1995 and 2002), and in-between the epidemic periods (1999 and 2000). The animals were caught by live-traps and identified by morphometric methods. The density of animals was estimated by counting the number of holes per 1000 m2. Sera of 311 patients with clinical signs and symptoms of hemorrhagic fever with renal syndrome (HFRS) were tested for the presence of antibodies reactive to the Dobrava, Puumala and Seoul viruses by using indirect immunofluorescence test (IIF), and IgG and IgM ELIS. Sera of 84 patients were tested using only IIF, and 227 sera were tested by IIF and -capture IgM ELIS tests. RESULTS During the epidemic years, the average monthly temperatures in February were by 4.3 times higher than the average temperatures during the nonepidemic years, which may have influenced the early reproduction of rodents and development of "mouse years". The rodents were identified as: Apodemus flavicollis (n = 139), Apodemus sylvaticus (n = 89), Apodemus agrarius (n = 4), Clethrionomys glareolus (n = 117), Sorex araneus (n = 5), Pytimus subterraneus (n = 23), Mus musculus (n = 1), Mycrotus arvalis (n = 1) and Rattus norvegicus (n = 2). Clethrionomys glareolus was predominant in the regions with the altitude higher than 1160 meters and Apodemus species in the regions with the altitude lower than 670 meters. The rodent population density changes seasonally and cyclically. During the epidemic years, the rodent population density was marked as very high, whereas during the nonepidemic years it was designated from low to moderate. Well-known natural hosts of Hantaviruses (A. flavicolis and C. glareolus) are most widely spread species of small rodents, and the increase in their population is closely related with outbreaks of epidemics of HVBS-a. Puumala virus caused HVBS-a in 49.84% (155/311); Dobrava virus in 23.15% (72/311) of cases, whereas Hantaviruses serotype was not identified in 27.00% (84/311) of cases. Infections caused by Puumala virus were more frequent than the infections caused by Dobrava virus during both epidemic and nonepidemic periods. The proportion of humans infected with Puumala and Dobrava viruses correlated with the number of natural hosts of Hantaviruses in the areas of HVBS outbreaks. The study of the prevalence of hantavirus antibodies in the populations of rodents and humans, which had been under way, should elucidate these relationships.

H. Selesković, S. Mulić, N. Basić, M. Hukić, A. Cickusic, M. Smajić

Background The deterioration of renal function is one of the most important prognostic predictors in lupus nephritis (LN). Objectives To determine glomerular filtration rate (GFR) in patients (pts) with LN and to clarify the characteristics of renal haemodynamics during the treatment of LN. Methods Thirty three clinically active pts with LN underwent glomerular filtration rate (GFR) clearance study by using radionucleotide compound Tc-99 m diethylamine-pentoacetic acid (DTPA). All pts had definite clinical evidence of active LN, including urinary and immunological abnormalities. Twenty of pts underwent renal biopsies (66%). GFR was studied in 30 female and 3 male pts (average 39 years and duration of disease average 5 years) in two occasions over an interval of 3 to 6 months during treatment with cotricosteroids (Cs) and cytotoxic drugs (Ct). Results GFR-DTPA clearance was in range from 11 to 157 ml/min, mean 87.42 ml/min (normal >100 ml/min), in repeated measurements it was in range from 13 to 147 ml/min, mean GFR (94.33) was slightly but not significantly better. We divided pts into four groups. First group of 12 pts had values in normal range, second group of 11 pts showed an improvement in GFR from mean 64.33 to 95.50 ml/min. Mean GFR second values from third group of 6 pts (18%) were as low as in first measurements (mean 51.83), and a fall in GFR (mean 106 to 70.25 ml/min) was notified only in 4 pts (12%) as a result of deterioration of glomerular function. Conclusion During the undergoing treatment over an interval of 3 to 6 month we kept GFR in pts with LN in normal range at 12 pts (37%). Average GFR increased significantly (p < 0.05) at 11 pts (33%). The treatment LN with Cs and Ct is useful for saving renal function, and could be able to prevent the progression LN to renal failure.

Å. Lundkvist, M. Hukić, J. Hörling, M. Gilljam, S. Nichol, B. Niklasson

Hantavirus infection was diagnosed serologically by μ‐capture IgM and IgG ELISAs in hemorrhagic fever with renal syndrome (HFRS) patients admitted to Tuzla Hospital, Bosnia‐Herzegovina. The results indicated that more than one hantavirus caused the outbreak. To address the question of which hantavirus serotypes were involved, sequentially drawn sera were analyzed by focus reduction neutralization test (FRNT) for antibodies against Puumala, Hantaan, Dobrava, and Seoul hantaviruses. The data revealed that acute‐ or early convalescent‐phase sera, even when drawn as late as 3 weeks after the onset of disease, could not be used for typing of the causative hantavirus; a significant number of these samples showed similar reactivity of neutralizing antibodies to several different hantavirus serotypes. Moreover, although several acute‐phase sera showed the highest FRNT titer to Hantaan virus, convalescent sera from these patients in all cases showed high specificity for Puumala or Dobrava viruses. This phenomenon, interpreted as a cross‐neutralizing primary antibody response, makes several earlier reports concerning causative agents of HFRS questionable. Serological examination of small rodents trapped in the endemic area identified Puumala‐ and Dobrava‐like virus infections. RT‐PCR and sequencing of rodent lung samples identified Dobrava virus in one yellow‐necked field mouse (Apodemus flavicollis). Cross‐FRNT data, using polyclonal rabbit antibodies, clearly confirmed Dobrava virus as a unique hantavirus serotype. In conclusion, the results revealed that both Puumala‐ and Dobrava‐like viruses caused HFRS in Bosnia‐Herzegovina, whereas no signs of Hantaan or Seoul virus involvement were found. J. Med. Virol. 53:51–59, 1997. © 1997 Wiley‐Liss, Inc.

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