Predrag Stojanović

The prevalence of Candida onychomycosis in Southeastern Serbia from 2011 to 2015.

Despite the increasing of onychomycosis caused by Candida spp., in referent literature, there is still data insufficiency about this nail infection. The objectives of this retrospective study were to determine epidemiological characteristics of Candida onychomycosis, the antifungal susceptibility of isolated species in vitro, and to compare the results of antifungal susceptibility testing with conducted treatment in period from 2011 to the end of March 2015. Out of 761 patients who were underwent clinical and mycological examinations, 137 had Candida species isolated from nails. The dominant species was Candida albicans (C. albicans) (36.59%) followed by C. parapsilosis (23.78%), C. krusei (9.76%), and C. guilliermondii (6.71%). Antifungal susceptibility in vitro testing showed good susceptibility to antimycotics, except C. krusei, which was resistance to fluconazole (FCZ) and isolates of C. tropicalis and C. glabrata which were dose dependent to itraconazole (ITZ) and fluconazole. Evaluation of medical histories determined that combined therapy, which included pulsed systemic regimen of ITZ with topical application of clotrimazole, had better clinical outcomes regarding the proscribed only topical application of clotrimazole. Multidisciplinary approach of dermatologists and mycologists is required in solving the problem of onychomycosis, which is the dominant nail disease.

High-density cervical ureaplasma urealyticum colonization in pregnant women

BACKGROUND/AIM Ureaplasma urealyticum, a common commensal of the female lower genital tract, has been observed as an important opportunistic pathogen during pregnancy. The aims of this study were to determine the degree of cervical colonization with U. urealyticum in pregnant women with risk pregnancy and in pregnant women with normal term delivery and to evaluate the correlation between high-density cervical U. urealyticum colonization and premature rupture of membranes (PROM) as well. METHODS This research was conducted on the samples comprizing 130 hospitalized pregnant women with threatening preterm delivery and premature rupture of membranes. The control group consisted of 39 pregnant women with term delivery without PROM. In addition to standard bacteriological examination and performing direct immunofluorescence test to detect Chlamydia trachomalis, cervical swabs were also examined for the presence of U. urealyticum and Mycoplasma hominis by commercially available Mycofast Evolution 2 test (International Microbio, France). RESULTS The number of findings with isolated high-density U. urealyticum in the target group was 69 (53.08%), while in the control group was 14 (35.90%). Premature rupture of membranes (PROM) occurred in 43 (33.08%) examinees: 29 were pPROM, and 14 were PROM. The finding of U. urealyticum > or = 10(4) was determined in 25 (58.14%) pregnant women with rupture, 17 were pPROM, and 8 were PROM. There was statistically significant difference in the finding of high-density U. urealyticum between the pregnant women with PROM and the control group (chi2 = 4.06, p < 0.05). U. urealyticum was predominant bacterial species found in 62.79% of isolates in the PROM cases, while in 32.56% it was isolated alone. Among the 49 pregnant women with preterm delivery, pPROM occurred in 29 (59.18%) examinees, and in 70.83% of pregnant women with findings of high-density U. urealyticum pPROM was observed. CONCLUSION Cervical colonization with U. urealyticum > or = 10(4) is more frequent in pregnant women with risk pregnancy than in pregnant women with normal term delivery. High-density cervical U. urealyticum colonization should be observed as a possible etiological factor for PROM.

Surveillance and characterization of Candida bloodstream infections in a Serbian tertiary care hospital

INTRODUCTION Candida spp. frequently cause hospital-acquired bloodstream infections (BSI) with a high mortality rate (up to 70%). We analyzed the frequency, infection characteristics, potential predisposing factors, susceptibility to antifungal drugs, biofilm production and other virulence characteristics of Candida spp. isolates obtained from a tertiary care hospital in Niš, Serbia, during a one year period. METHODS Medical histories, characteristics of isolated strains and drug susceptibility, as well as the effect on the function of isolated macrophages and other virulence features were evaluated. The obtained results were subjected to students t-test and multivariate statistical analyzes. RESULTS Herein we report an annual incidence of 3.65 cases of C. albicans, C. lusitaniae and C. lipolytica infections per 105 population. Out of eight isolated strains, two (25%) were shown to be strong biofilm producers, one (12.5%) caused hemolysis on blood agar and in two (25%) cases macrophages were able to completely eliminate the yeast colonies. Chronic kidney disease, diabetes, malignant and other diseases were present in 37.5, 62.5, 50 and 75%, respectively, in the study group. All patients with Candida BSI received antifungal therapy (amphotericin B), however, hospital mortality was observed in 25% of patients. CONCLUSIONS Evaluation of local Candida epidemiology, antifungal susceptibility and virulence factors, as well as personalized patient risk factors are important for the surveillance of Candida BSI, especially in intensive care unit patients and may contribute to the improved options and outcome for patients with Candida BSI.

Asymptomatic carriers of clostridium difficile in serbian population

The aim of the research was to determine the intestinal carriers of C. difficile in different human population groups in Serbia. The research enrolled 877 persons with formed stools: (newborn children in maternity hospitals for up to two weeks old) (23), group A; children aged from two weeks to two years (121), group B; children aged two to 10 years (54), group C, healthy individuals aged 10 and over (516), group D; patients hospitalized for at least 48 hours (100), group E; staff of the Clinical Center in Nis, Serbia, (63), group F. The toxins A and B of C. difficile were detected by ELISA-ridascreen Clostridium difficile Toxin A/B (R — Biopharm AG, Darmstadt, Germany). The toxin A of C. difficile was detected using ColorPAC Toxin A test (BectonDickinson, New Jersey, USA). Out of the total number of persons (877), the carriers of certain types of toxin-producing strains of C. difficile were distributed as: 6.04% (A−/B−), 1.83% (A+/B+) and 0.11% (A−/B+). In most groups (5/6), the dominance of non-toxigenic (A−/B−) isolates was established, with the rate of carriers 1.75 – 30.43% depending on the group. Toxigenic isolates were prevalent only in the group F in relation to non — toxigenic (7.94% versus 4.76% of persons). In other groups, the carriers of toxigenic strains ranged from 0.00 – 17.45%. The presence of asymptomatic intestinal carriers of C. difficile in the human population, indicate the possible reservoirs and sources of infection.

Superficial mycoses in the Nis region, Southeast-Serbia

The aim of the study is to investigate the most frequent cause of superficial mycoses in patients from the territory of city Niš Southeast Serbia in the period from 1998 to 2010. A total of 3223 samples from 2887 patients with suspected dermatomycoses were examined. Superficial mycoses were diagnosed using standard microbiology techniques (conventional microscopy and cultivation). Dermatophytes were determined on the basis of their macroscopic and microscopic morphological and morphometric characteristics. Morphometric characteristics were obtained by Laboratory Universal Computer Image Analysis system (Lucia M, 1996). Species of genus Candida were identified using the test of production of germ tube in sera, by growth on comertial chromatogen medium (Chromotogenic Candida, Liofichem/Bacteriology products, Italy) and by using Auxacolor TMBioRad, France. The results were elaborated with the statistical method of descriptive and quantitative analysis (SPSS 14.0 for Windows 2003). The prevalence of superficial mycoses was 25,1%. Dermatophytes were identified in 67.6% of all positive cultures. Microsporum canis was the most prevalent (50.3%) dermatophyte isolated, followed by Trichophyton metagrophytes var. mentagrophytes (35.4%). Yeast genus Candida has become a more frequent cause of superficial fungal infection since 2001. and C. albicans was the dominant yeast (61.1%).

Clinical importance of Clostridium difficile finding in hospitalized patients

INTRODUCTION Clostridium difficile infections predominatelly occur among hospitalized patients. The aim of this study was to evaluate the importance of finding the isolate of Clostridium difficile cultured from the stool of hospitalized patients. MATERIAL AND METHODS Material consisted of 100 patients with at least one liquid stool samples and control group with form stool. Every patient spent minimum 48h in hospital before the sampling. The material was immediately cultured on mediums for isolation of enteric pathogens, and on selective CCFA medium (Biomedics) for Clostridium difficile in anaerobic condition. Diagnosis of Clostridium difficile toxin in stool samples was achieved by ELISA-RIDASCREEN Clostridium difficile Toxin A/B test (R-Biopharm). RESULTS One-hundred forty one stool samples of patients in Clinical Centre Nis were cultivated and examined for C. difficile. The bacteria was isolated in seven patients from the clinical group. In four (57.14%) patients, the presence of C difficile toxin in stool was established The bacteria was diagnosed from the stool samples of five patients from the control group, but the toxin was not found in their stool samples. DISCUSSION The results performed at the Institute for Public Health Nis are in accordance with previously published results that all patients with positive findings of Clostridium difficile toxin in stool samples were on antibiotic treatment longer than 14 days. By analysing the patients stay in hospital and duration of antibiotic treatment, we observed the statistically significant difference in findings between the patients with CDAD and the patients from the control group with positive bacteria. CONCLUSION The study confirms the importance of finding Clostridium difficile associated disease in four (4%) hospitalized patients.

Diarrhoea caused by Clostridium difficile in patients with postoperative subhepatic abscess

BACKGROUND Toxigenic strains of Clostridium difficile in the majority of cases cause disease of the intestinal tract of hospitalized patients. For a long time, Clostridium difficile was considered to produce both types of toxins (A+/B+ strain), however, the investigations conducted in the last ten years point to the existence of clinically significant isolates which produce only toxin B, i. e. toxin A negative/toxin B positive (A-/B+ strain) Clostridium difficile. CASE REPORT We presented the case of a patient admitted to the Surgery Clinic, Clinical Center Nis due to the presence of calculus in the ductus choledochus. Twenty-four hours after the surgical intervention for calculus removal, the first signs of the operative wound infection began to appear. In the course of infection treatment, different antibiotics were administered (cefuroxine, ciprofloxacin, vancomycin, imipenem). After making etiological microbiological diagnosis and application of antibiotics according to antibiogram results, the signs of the operative wound infection began to withdraw, but the patient reported the abdominal pain and liquid stools with traces of blood (up to 17 stools per day). By microbiological examination, Clostridium diffidile was cultivated and the presence of toxin B was detected in the stool samples. The patient was sent to the Clinic for Infectious Diseases, where the causal therapy of mitronidazol was administered. Liquid and electrolytes were made up by substitution therapy. After the eight-day-treatment, the patient felt much better, and diarrheas stopped on the 10th day of the therapy application. CONCLUSION Our results have shown that toxingen strains Clostridium difficile are present in our country so this bacterium sort have to be considered in differential causal diagnosis of diarrhoea syndrome. Considering that it can cause difficult form of the disease, it is an obligation to establish the presence of some toxins of Clostridium difficile in stool samples of patients and/or production of some toxins in liquid culturate of isolates to provide data for the presence of strains which produce only toxin B.

Contribution to the History of Serbian Medicine: Ten Years from the Introduction of Clostridium difficile into Microbiological Diagnostics in Serbia

Summary In Serbia, the first isolates of C. difficile were isolated in the Public Health Institute (PHI), Center for Microbiology in Niš, at the end of 2005. The National Reference Laboratory for Anaerobic Infections (NRLA) in PHI Niš confirmed the toxigenic strains that caused the first three registered hospital epidemics in Serbia, in 2006 at the Clinic of Neurology, Clinical Center Niš, in 2007 at the Clinical Center of Vojvodina in Novi Sad, and in 2009 in the General Hospital in Požarevac. In 2014, C. difficile species were isolated for the first time from 175 environment samples in the research studies which were conducted in NRLA of PHI Niš. In the samples of soil taken from the ground within the Clinical Center Niš, those taken from the parks at the territory of the Municipality of Niš, samples of mud and sand around the illegal sewage systems at the territory of the Municipality of Niška Banja, a small number of bacteria C. difficile producing the toxins (A+B+) as well as non-toxigenic isolates (A−B−) were found. Results of the first epidemiological investigations of cases of diarrhea associated with prior antibiotic treatment applied in hospitalized patients in a number of health centers in our country, microbiological investigations done in the Public Health Institute, valuable discussions at professional and scientific meetings influenced the general attitude that isolation and identification of C. difficile and/or detection of toxin produced by this bacteria should be part of the routine work in the Serbian microbiological laboratories.