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Background Autochthonous human infections with Strongyloides stercoralis have been well documented in many European regions. By exploring patients’ data, we aimed to find elements for its current endemicity in Croatia. Methods This retrospective descriptive study analysed epidemiological and clinical data of patients treated for strongyloidiasis from January 2010 to May 2019 at a teaching hospital in Zagreb, Croatia. The diagnosis was made by direct methods using light microscopy and/or serology. Results Among 65 patients with strongyloidiasis, 60% were men, and 78.5% were 50–79 y of age. The sensitivity of the examination of three stool samples after concentration, saline provocation and serology was 26.2, 80.7 and 86.2%, respectively. Clinical presentation included asymptomatic patients with eosinophilia (41.5%), chronic symptomatic infection (33.8%), acute infection (18.5%) and hyperinfection (6.2%). Twenty patients (30.8%) were immunosuppressed; among four of whom developed hyperinfection, two died. Initially 71.7% of patients were treated with albendazole and 13.3% with ivermectin, with an equal parasitological cure rate (72.2% and 75%; p=0.09). In 11 patients, acute infection was autochthonous in its origin. Conclusions Strongyloidiasis is currently endemic in Croatia and immunosuppressed travellers to this region should be advised to take precautions. Patients undergoing immunosuppression and organ donors from Croatia should be screened.

Review The neglected zoonosis cystic echinococcosis affects mainly pastoral and rural communities in both low-income and upper-middle-income countries. In Europe, it should be regarded as an orphan and rare disease. Although human cystic echinococcosis is a notifiable parasitic infectious disease in most European countries, in practice it is largely under-reported by national health systems. To fill this gap, we extracted data on the number, incidence, and trend of human cases in Europe through a systematic review approach, using both the scientific and grey literature and accounting for the period of publication from 1997 to 2021. The highest number of possible human cases at the national level was calculated from various data sources to generate a descriptive model of human cystic echinococcosis in Europe. We identified 64 745 human cystic echinococcosis cases from 40 European countries. The mean annual incidence from 1997 to 2020 throughout Europe was 0·64 cases per 100 000 people and in EU member states was 0·50 cases per 100 000 people. Based on incidence rates and trends detected in this study, the current epicentre of cystic echinococcosis in Europe is in the southeastern European countries, whereas historical endemic European Mediterranean countries have recorded a decrease in the number of cases over the time. Key messages: - This study aims to shed light on the unrecognised incidence of cystic echinococcosis in Europe, unveiling its epidemiological effect by providing a quantitative measure of number, incidence, and trends of human cases documented within the period 1997–2021; - Since human cystic echinococcosis cases are generally under-reported and data have uncertainty (partly due to misdiagnosis), data provided in this study should be considered as a conservative estimate of the real impact of this zoonotic infection historically occurring in Europe; - For the years 2017–19, we identified a total number of cystic echinococcosis cases four-fold higher than for The European Surveillance System (TESSy) data; - Decreasing trends have been recorded in most southern Mediterranean and some eastern European countries, where cystic echinococcosis has traditionally been highly prevalent; - Increasing trends have been identified in some eastern and southeastern European countries but, unexpectedly, also in most non-endemic countries of northern and western Europe; - Based on incidence and trends from 2017–19, the current epicentre of cystic echinococcosis in Europe is represented by the Balkan Peninsula; - Cystic echinococcosis in Europe remains a relevant public health issue and findings from this study should be used to support the planning of surveillance and control programmes in Europe according to the WHO 2021–2030 roadmap for neglected tropical diseases. This research was funded by the MEmE project from the EU’s Horizon 2020 Research and Innovation programme under grant agreement number 773830: One Health European Joint Programme. This work was also partially supported by research funding (grant PRG1209) from the Estonian Ministry of Education and Research info:eu-repo/semantics/publishedVersion

The predominance of the genus Corynebacterium in the healthy male urogenital system contributes to the resident microbiome of not only the distal urethra, but potentially the proximal urethra and urinary bladder as well. However, for certain species in this genus, pathogenic potential was described, and the salient representative is Corynebacterium glucuronolyticum (C. glucuronolyticum) implicated in cases of urethritis and prostatitis in men. Nonetheless, some still question whether C. glucuronolyticum can actually be considered pathogenic or rather just a commensal species fortuitously isolated in patients with urogenital symptoms and/or syndromes. Although pathogen/commensal dichotomy is not always clear-cut, we hypothesize that specific genetic markers may expose C. glucuronolyticum as a convincingly pathogenic Corynebacterium. More specifically, characteristic pathogenic gene constellation inherent to this species (most notably the presence of specific sortase/SpaA-type pili gene clusters, but also the augmentative role of type VII secretion system) may significantly facilitate host tissue adhesion, with subsequent suppression/evasion of the immune response and acquisition of vitally important nutrients. Consequently, these genetic markers differentiate C. glucuronolyticum from its commensal counterparts, and give this species a pathogenic facet, which can be even further influenced by the Allee effect. In this paper we also propose a specific methodological approach on how to analyze C. glucuronolyticum epithelial colonization capacity and explore inceptive host cell-pathogen interactions that manipulate host environment and immune responses. This entails moving from approaches based primarily on overall homology of primary sequences towards specific structure-function studies to precisely evaluate all stakeholders involved in pili assemblage, cell adhesion and the expression of other virulence traits. In the era of high precision medicine, the hypothesized roles of C. glucuronolyticum adhesion systems in both virulence and nutrient acquisition may also reveal promising targets for future drug developments.

Background: Previous studies suggested a complex association between Toxoplasma gondii (TG) infection and host lipid metabolism. Both TG infection and metabolic disturbances are very common in patients with schizophrenia, but this relationship is not clear. Methods: In this cross-sectional study, we evaluated the association between TG seropositivity, serum lipid levels, body mass index (BMI) and metabolic syndrome (MetS) in 210 male inpatients with schizophrenia. Results: In our sample of schizophrenia patients, with the mean age of 43.90 ± 12.70 years, the rate of TG seropositivity was 52.38% and the prevalence of MetS was 17%. Patients with the TG antibodies had lower serum triglyceride levels and body weight compared to TG seronegative patients, despite having more frequently received antipsychotics (clozapine, olanzapine risperidone and quetiapine), which are well known to induce weight gain and metabolic abnormalities. However, the only significant change in metabolic parameters, observed in TG seropositive patients with schizophrenia, was decreased serum triglyceride to high-density lipoprotein cholesterol (HDL-C) ratio. No associations were observed between TG seropositivity and serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and glucose levels, waist circumference, BMI and the rate of MetS. Conclusion: This is the first report of the association between TG infection and decreased serum triglyceride to HDL-C ratio in a sample of carefully selected men with chronic schizophrenia. Keywords: Toxoplasma gondii infection, Schizophrenia patients, Serum lipid indices, Body mass index, Metabolic syndrome

Genital warts (also known as condylomata acuminata) caused by the human papillomavirus (HPV) represent one of the most common sexually transmitted infections. Although they are usually found in the outer genital region, a small proportion of men can present with (often unrecognised) intraurethral warts, generally limited to the distal urethra and urethral meatus. This poses a treatment challenge not adequately addressed by the current guidelines. Here, we present two cases of low-risk HPV-positive patients with protruding and non-protruding condylomata acuminata of the distal urethra, which were treated successfully by using two different topical regimens (ie, a combination of policresulen and imiquimod for one patient and 5-fluorouracil monotherapy for the other). Although this type of management results in lower rates of tissue destruction and complications and may be given preference as an initial therapeutic option, additional prospective comparative clinical studies are needed to elucidate its potential in similar cases.

Corynebacterium glucuronolyticum (C. glucuronolyticum) is a rare isolate that is only recently being acknowledged as a potential urogenital pathogen. The bibliographical references on this bacterial species are scarce, and its influence on all semen parameters was hitherto unknown - therefore, the aim of this study was to evaluate its effects on a range of sperm quality parameters. A prospective approach to compare semen parameters before and after treatment was used in this study. C. glucuronolyticum in semen specimens was identified using analytical profile index biotyping system (API Coryne) and additionally confirmed by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS), with the determination of antimicrobial susceptibility by Kirby-Bauer method. Semen analysis was performed according to the criteria from the World Health Organization (with the use of Tygerberg method of sperm morphology categorization). Very strict inclusion criteria for participants also included detailed medical history and urological evaluation. From a total of 2169 screened semen specimens, the inclusion rate for participants with C. glucuronolyticum that satisfied all the criteria was 1.01%. Antibiogram-guided treatment of the infection with ensuing microbiological clearance has shown that the resolution of the infection correlates with statistically significant improvement in the vitality of spermatozoa, but also with a lower number of neck and mid-piece defects. Parameters such as sperm count, motility and normal morphology were not affected. In addition, susceptibility testing revealed a trend towards ciprofloxacin resistance, which is something that should be considered when selecting an optimal treatment approach. Albeit it is rarely encountered as a monoisolate in significant quantities, C. glucuronolyticum may negatively influence certain sperm parameters; therefore, it has to be taken into account in the microbiological analysis of urogenital samples.

Although trichinellosis represents one of the most significant global foodborne zoonotic diseases, human seroprevalence studies are scarce. Due to its endemicity in Croatia, the aims of this study were to establish the general prevalence of Trichinella infection in Croatian individuals presenting with eosinophilia during a 5-year period, and to assess the value of such screening endeavours for early detection/differentiation of outbreaks. A total of 1342 serum samples were collected from male and female subjects presenting with peripheral blood eosinophilia between 2013 and 2017, and tested for Trichinella IgG antibodies by employing commercial qualitative ELISA and Trichinella Excreted/Secreted (E/S) Western blot methods. The overall positivity was 3.65%, ranging from 0.49% to 1.50% between 2013 and 2016, but rising to 10.98% in 2017; such ten-fold increase in seroprevalence forecasted the epidemic situation in Croatia. Overall the prevalence of the infection rose with the increasing age and the rate was highest among those older than 40 years of age (p=0.003), without any significant sex-based differences (p=0.438). Considering the early appearance of eosinophilia in the infected individuals, this type of systematic screening can be seen as an additional epidemiological tool to unveil the trichinellosis outbreak in a timely manner.

Aim The aim of the study was to determine in vitro synergy and postantibiotic effect of colistin alone and combined with meropenem or vancomycin against Enterobacteriaceae producing multiple carbapenemases; combinations of two metallo-β-lactamases (MBL) or MBL with OXA-48. Colistin-resistant strain positive for OXA-48 was also included in the study. Methods The antibiotic susceptibility was tested by broth microdilution method. Synergy was tested by chequerboard, time-kill and 2-well method. PAE was determined by viable counting. Results The chequerboard analysis revealed synergy for colistin combination with meropenem in all isolates with FICI values ranging from 0.12 to 0.24. FICI values for combinations with vancomycin were below 0.5 indicating synergy in two out of four isolates. K. pneumoniae 609815 positive for OXA-48 and colistin resistant showed the most pronounced and consistent synergy effect with meropenem in both chequerboard and time-kill method. Synergy effect in time-kill curves, was observed for K pneumoniae 145846 with two MBLs and colistin resistant K. pneumoniae 609815 positive for OXA-48, with both combinations including meropenem and vancomycin. Colistin alone exhibited short postantibiotic effect (PAE) against all tested isolates. Meropenem markedly prolonged the PAE in two isolates in contrast to vancomycin which did not demonstrate significant effect on the duration of PAE. Conclusions The synergy effect and the duration of PAE was strain and antibiotic dependent but not related to the resistance gene content.

Strongyloides stercoralis is an intestinal nematode that causes chronic gastrointestinal infection. In immunosuppressed patients this persistent disease can lead to disseminated disease and/or hyperinfection that are life threatening conditions. We present a fatal case of S. stercoralis hyperinfection and the appropriate treatment for immunosuppressed patients is discussed., Strongyloides stercoralis je crijevna nematoda koja uzrokuje kroničnu gastrointestinalnu infekciju. Kod imunokompromitiranih bolesnika ova perzistentna bolest može dovesti do diseminirane bolesti i/ili hiperinfekcije koje su životno ugrožavajuće. Prikazujemo slučaj hiperinfekcije S. stercoralis sa smrtnim ishodom te odgovarajuće liječenje kod imunokompromitiranih bolesnika.

Objectives The main objective of this study is to determine the prevalence of trichomoniasis in men with and without symptoms of urethritis, with concomitant analysis of sociodemographic and behavioural specificities of both groups. Also, the objective is to evaluate laboratory methods used in the diagnostics of this parasitic disease. Methods A total of 500 men with and 200 without urethritis symptoms were included in the study. Every respondent filled out a questionnaire asking for some general data, specific information about habits, sexual behaviour and symptoms. Sediment of first void urine was analysed by wet mount microscopy, cultivation in Diamond9s medium and real-time PCR. Results In the symptomatic group, Trichomonas vaginalis infection was documented in 2.4% of respondents by wet mount microscopy, in 4.8% by cultivation and in 8.2% by real-time PCR. In the asymptomatic group, infection was proven using the same methods in 1.0%, 1.5% and 2.0% of the respondents, respectively. Trichomoniasis prevalence was statistically significantly higher in the respondents manifesting urethritis symptoms when cultivation (χ 2 =4.20, p=0.041) and real-time PCR (χ 2 =9.20, p=0.002) were used. Several epidemiological risk factors were identified, and greater sensitivity of real-time PCR was found in comparison with microscopy and culture. Conclusions Trichomonas infection was statistically more frequent in men with urethritis syndrome. Assuming that the samples found positive by any laboratory technique are truly positive, it can be concluded that the real-time PCR showed the greatest sensitivity of all the methods used in this study.

In order to identify the groups at risk of sexually transmitted diseases (STDs), we assessed the sociodemographic profiles of men testing for STD, their sexual habits, and the results of microbiological analysis. During a three-year period, a total of 700 men older than 18 years of age completed the questionnaire regarding sociodemographic and sexual behavior. Urethral swabs were taken for microbiological analysis. Thirty-three percent of respondents reported not using condoms. Those that do not use condoms were predominantly less educated, unmarried but in steady relationships, employed, with children, and smokers. Alcohol or drug usage before sexual intercourse was disclosed by 21.4% of respondents, and 10.3% respondents reported sexual intercourses with commercial sex workers. Finally, 24.0% respondents reported sexual relations abroad. In 28.1% of subjects, one or more pathogens were observed in urethral swabs. The most commonly diagnosed microorganism was Ureaplasma urealyticum, followed by Chlamydia trachomatis, Mycoplasma hominis, Trichomonas vaginalis, and Neisseria gonorrhoeae. This study identified several factors that may contribute to the general risk of STD transmission, which will serve to better understand the transmission dynamics and implementation of adequate prevention programs.

Background: Chlamydia trachomatis (C. trachomatis) is the most common bacterial agent of sexually transmitted infections around the world, but susceptibility testing of this pathogen is rarely pursued due to its intracellular niche. The principal aims of this research were to determine in vitro sensitivity profile of urogenital chlamydial strains isolated from Croatian patients and to compare obtained concentration values of different antimicrobial drugs mutually and with the literature. Methods: Forty strains of C. trachomatis isolated during 2010 - 2012 at the National Reference Laboratory for Chlamydia and two reference strains were subjected to susceptibility testing in 96-well microtiter plates containing McCoy cell monolayers. Minimal inhibitory concentration (MIC) and minimal chlamydicidal concentration (MCC) were determined for azithromycin, doxycycline, and levofloxacin. Briefly, strains were inoculated on McCoy cells, followed by addition of serially diluted antimicrobial drugs. Upon incubation, growth of C. trachomatis was detected using fluorescein-conjugated antibody to the lipopolysaccharide genus antigen under the inverted fluorescent microscope. Results: All chlamydial strains were susceptible to the antibiotics tested (MIC < 4 μg/mL), thus the pattern of homotypic or heterotypic resistance has not been found. MCC values were equal or 1 - 5 dilutions higher than MIC values. Statistically significant differences in the effectiveness of antimicrobial agents in vitro have been proven. Significant correlation has been found for MCCs in the case of two antimicrobial pairs: azithromycin and levofloxacin, and doxycycline and levofloxacin. Comparison of medians for different clinical samples did not reveal any significant difference. Conclusions: Although resistant strains have not been found in this study, several literature reports of unsuccessfully treated genitourinary infections caused by C. trachomatis require our alertness for possible discovery of resistant strains. Considering the overall antibiotic burden worldwide, pursuing this kind of research is crucial in order to detect possible decreased susceptibility (or even resistance) of chlamydial strains, despite the laborious and time-consuming methodology.

Hepatitis E has become an emerging infection in many European countries. We analysed the prevalence of hepatitis E virus (HEV) infection in selected population groups in Croatia. Overall HEV IgG seropositivity was 5.6%, while 1.9% participants showed IgM antibodies suggestive of recent infection. No IgM-positive sample was positive for HEV RNA. HEV IgG antibodies were most prevalent in alcohol abusers (8.9%) and war veterans (8.6%), compared with 6.1% among injecting drug users and 2.7% in healthcare professionals. No individual with high-risk sexual behaviour tested HEV seropositive. HEV IgG positivity increased significantly with age from 1.8% to 2.3% in individuals younger than 40 years to 11.3% in individuals older than 50 years (P = 0.023). The mean age of HEV-positive participants was significantly higher than that of HEV-negative participants (50.9 ± 11.8 years versus 41.2 ± 11.8 years, P = 0.008). Seroprevalence rates were significantly higher in residents of suburban and rural areas compared with residents of urban areas (14.5% versus 2.5%, P = 0.003). Additionally, an increasing prevalence of HEV IgG antibodies was observed from 1.8% in participants living in families with two household members to 12.1% in those living with more than four members (P = 0.046). Gender, marital status, educational level, sexual orientation, source of drinking water, history of blood transfusions, surgical procedures, tattooing and travelling were not associated with HEV seroprevalence. Logistic regression showed that living in suburban/rural areas was the main risk factor for HEV seropositivity (OR = 6.67; 95%CI = 1.89-25.0; AOR = 7.14, 95%CI = 1.89-25.0).

Toxoplasma gondii je intracelularni parazit svih stanica s jezgrom, ali poseban afinitet ima prema endotelu i živčanom tkivu. U posljednje vrijeme infekcija parazitom Toxoplasma gondii povezuje se s razvojem pojedinih psihijatrijskih poremećaja. Jedan od najčešće proučavanih psihijatrijskih poremećaja na ovom polju jest shizofrenija. Međutim, postoje podatci i o povezanosti toksoplazme s drugim psihijatrijskim poremećajima kao što su opsesivno-kompulzivni poremećaj, te poremećaji raspoloženja, bipolarni poremećaj i depresivni poremećaj. Dobro je poznato kako bolesnici sa shizofrenijom imaju abnormalnu funkciju neurotransmitora, osobito dopamina, glutamata i GABA-e. Mogući mehanizam pomoću kojeg Toxoplasma gondii utječe na razinu dopamina je taj da Toxoplasma gondii posjeduje 2 gena koja kodiraju enzim tirozin hidroksilazu, važan enzim u sintezi dopamina. Toxoplasma gondii u tahizoitnom obliku sklona je izazvati intenzivan imunološki odgovor i posljedično oslobađanje velikog broja citokina. Upravo bi ruptura ciste ispunjene tahizoitima mogla biti potencijalni etiološki čimbenik razvoja psihijatrijske patologije. Izloženost infekciji toksoplazmom povezana je s kognitivnim deficitom u shizofrenih bolesnika, ali i u ostaloj populaciji. S obzirom na porast broja istraživanja na ovom polju medicine i na temelju rezultata koji su objavljeni, postoji vjerojatnost kako je infekcija parazitom Toxoplasma gondii povezana s patološkim promjenama u mozgu. No, na brojna pitanja još uvijek nisu pronađeni odgovori., Toxoplasma gondii is an obligate intracellular parasite which usually invades endothelial cells and neural tissue. Over the past years there has been an increasing interest in the possibility of an association of Toxoplasma gondii infection and psychiatric disorders. Schizophrenia is the most frequently analyzed in this field. However, Toxoplasma gondii infection can also be linked with other psychiatric disorders, such as obsessive-compulsive disorders and mood disorders, including bipolar disorder and depression. It is well known that individuals with schizophrenia have dysfunctional neurotransmitters, especially dopamine, glutamate and GABA. A possible mechanism by which Toxoplasma influences the function of the dopaminergic system is that the genome of Toxoplasma contains two genes for the enzyme tyrosine hydroxylase, the key enzyme for the synthesis of dopamine. Tachyzoites induce more intense inflammatory cytokine-mediated response. Cyst rupture in the brain could be related to the onset of mental diseases. Toxoplasma gondii exposure has been associated with cognitive impairment in both schizophrenic and non-schizophrenic subjects. Due to the growing interest in this subject in medicine, there the association between the exposure to Toxoplasma and neurodevelopment disorders can be considered likely. However, many of these questions are still unanswered.

During 2005-2009, a seroepidemiological study was carried out in Croatia to define the population susceptible to common TORCH agents among pregnant and non-pregnant women of childbearing age. The IgG seroprevalence was 29.1% forT. gondii, 94.6% for rubella, 75.3% for cytomegalovirus (CMV), 78.7% for herpes simplex virus type 1 (HSV-1), and 6.8% for HSV-2. Acute toxoplasmosis and CMV infection (positive IgM antibodies with low IgG avidity) were documented in 0.25% and 0.09% women, respectively. IgM prevalence was 1.2% for both HSV-1 and HSV-2. None of the participants showed acute rubella infection. Seropositivity to T. gondii and HSV-2 varied significantly between age groups (p = 0.001 and p = 0.036, respectively). Women residing in rural regions showed a significantly higher seroprevalence rate for T. gondii, CMV, and HSV-1 than urban women (T. gondii: 44.0% vs. 25.4%, p < 0.001 ; CMV: 85.0% vs. 73.1%, p = 0.018 ; HSV-1: 86.0% vs. 76.4%, p = 0.041).

The aim of this study was to compare the sensitivity and specificity of six different beta-lactam antibiotics using five phenotypical tests for detection of extended spectrum beta-lactamases (ESBLs) based on synergism of beta-lactam antibiotics and clavulanate. Experiments were performed on a set of 80 Klebsiella pneumoniae strains and 105 Escherichia coli strains with previously characterized ESBLs (SHV, TEM and CTX-M). ESBLs were detected by five different phenotypical methods: MIC (minimum inhibitory concentration) determination of beta-lactam antibiotics with and without clavulanate, double-disk synergy test (DDST), inhibitor-potentiated disk-diffusion test (IPDDT), CLSI-Clinical and Laboratory Standard Institution (former NCCLS) combined-disk-test, and modified MAST-disk-diffusion test (MAST-DD-test). Seven antibiotics were tested as indicators of ESBL production: ceftazidime, cefotaxime, ceftriaxone, aztreonam, ceftibuten, cefpodoxime and cefepime. Ceftazidime and aztreonam were the best indicators for SHV-5, SHV-12 and TEM beta-lactamases whereas cefotaxime and ceftriaxone were the most sensitive in detection of SHV-2 and CTX-M beta-lactamases in DDST, IPDDT and CLSI test. MIC determination of beta-lactam antibiotics with and without clavulanate was the most sensitive method. DDST was the least sensitive test. Double-disk synergy test, which is the most frequently used test for detection of ESBLs in routine laboratories, was the least sensitive independently of the indicator antibiotic. Since MIC determination is a very laborious and time consuming method, we would recommend the NCCLS combined disk test or IPDD test for detection of ESBLs in routine laboratories with 5 mm zone augmentation breakpoint.

Gastrointestinal manifestations of chlamydial infection are frequent, yet not always recognised. One of the common entities is proctitis, especially prevalent amongst men who have sex with men (MSM). Likewise, some enterohepatic Helicobacter species have also been associated with proctitis, namely Helicobacter (H.) cinaedi and H. fennelliae. It is well established that Helicobacter species have general and specific mechanisms for innate immune evasion and suppression, and can affect intestinal homeostasis. Here it is proposed that their presence in the rectum might facilitate the development of Chlamydia trachomatis proctitis, where they could act as cofactors in initial infection and progression of the disease.

IntroductionPrevious studies suggested that patients with schizophrenia had an increased prevalence of antibodies against toxoplasma gondii (TG) and that those seropositive patients had higher symptom severity. However, there is no data on the relationship between treatment-resistant schizophrenia (TRS) and TG seroprevalence.ObjectivesTo determine the association between TRS and TG seropositivity, and to further investigate the relationship between TG seropositivity and different clinical features of schizophrenia.MethodsIn this cross-sectional study, we included 210 male inpatients with schizophrenia. TG seropositivity was determined by ELFA assay. Treatment-resistance was defined as a failure of at least 2 adequate anti-psychotic trials. Data were analyzed using χ2 test or Mann–Whitney test.ResultsThe rate of TG seropositivity in the entire sample was 52.3%, whereas 47.6% of patients met the definition for treatment-resistance. Seropositive patients had twice the rate of treatment–resistance compared to seronegative patients (63.6% vs. 30.0%, P < 0.0001). Moreover, in the seropositive group, the patients were older (47.6 ± 12.2 vs. 39.81 ± 12.01 years, P < 0.0001), had higher number of previous hospitalizations (13.9 ± 11.7 vs. 9.6 ± 8.5, P = 0.0073), and increased Calgary depression scale for schizophrenia (CDSS) total score (7.8 ± 4.5 vs. 6.3 ± 3.8, P = 0.012). There were no differences between the groups in the age of disease onset, smoking, positive and negative syndrome scale (PANSS) total, positive and negative scores, and the life-time history of suicide attempts.ConclusionsOur results support the hypothesis that TG seropositivity might contribute to treatment-resistance in schizophrenia, at least in male patients.Disclosure of interestThe authors have not supplied their declaration of competing interest.

Introduction: Corynebacterium glucuronolyticum is a non‐lipophilic coryneform species and a rare isolate from male patients with genitourinary tract infections. This organism is typically associated with urethritis or prostatitis syndrome, although it can also be isolated from other sites. Case Presentation: We describe a case of urethritis in a 24‐year‐old Croatian male caused by a strain of C. glucuronolyticum showing resistance to ciprofloxacin in an agar dilution susceptibility test procedure. The same drug was used as an empirical treatment before microbiological evaluation was conducted. Upon identification of the organism, the patient was successfully treated with doxycycline, which resulted in complete regression of symptoms and lack of growth of the organism in culture. Conclusion: Although uncommon detection of this organism hinders research efforts, C. glucuronolyticum should always be regarded as a possible cause of urogenital infections in male patients. In addition, antibiotic susceptibility testing should be performed due to possible resistance.

Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program is a longitudinal global surveillance study to monitor in vitro data on microbial susceptibility in centers that prescribe meropenem. Results of the six years period (2002-2007) for the antimicrobial efficacy of meropenem compared to other broad-spectrum agents against Gram-negative and Gram-positive isolates collected at pediatric intensive care units of the University Hospital Center Zagreb in Croatia were reported.A total of 110 Gram-negative and 43 Gram-positive pathogens from pediatric specimens were tested. The minimum-inhibitory concentrations (MICs) were determined by broth microdilution method according to CLSI.There was no resistance to either imipenem or meropenem observed for Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. High resistance rates of K. pneumoniae to ceftazidime and gentamicin (50%) are a raising concern. Pseudomonas aeruginosa was the most resistant Gram-negative species with two (12%) of the strains resistant to meropenem, three (18%) to imipenem, 10 (47%) to gentamicin and six (35%) to piperacillin/tazobactam and ciprofloxacin. According to our results meropenem remains an appropriate antibiotic for the treatment of severe infections caused by Gram-negative bacteria in pediatric population.The results indicate that meropenem has excellent potency and spectrum of activity despite being prescribed for a long time for the treatment of seriously ill patients, and still appears to be a reliable option for the initial empirical therapy of serious nosocomial infections in children. However, later studies have shown the emergence of carbapenem-resistant Gram-negative bacteria after 2008.

Although Chlamydia trachomatis resistance is not of great concern due to its excellent sensitivity to the currently recommended first-line antibiotics (azithromycin and doxycycline), clinical treatment failures have been reported and some of them were linked to laboratory proved resistance. The aim of this study was to determine in vitro susceptibility to azithromycin and doxycycline for 24 urogenital chlamydial strains isolated in Croatia-a country with the highest consumption of azithromycin in Europe and with very high antibiotic prescription rates. Fourteen isolates from cervical swabs, nine from male urethral swabs, and one isolate from expressed prostatic secretion were tested in McCoy cell culture system. All strains were susceptible to azithromycin and doxycycline with minimal inhibitory concentration for azithromycin and doxycycline ranging from 0.064 to 0.125 μg/mL and 0.016 to 0.064 μg/mL, and minimal chlamydicidal concentration ranging from 0.064 to 2.0 μg/mL and 0.032 to 1.0 μg/mL, respectively. Since we still lack information on whether C. trachomatis is evolving in vivo in response to antibiotic selection pressure, this kind of surveillance for resistance is essential in detecting shifts in antimicrobial susceptibilities.

Dengue je akutna virusna bolest koju na čovjeka prenose komarci roda Aedes (Ae. aegypti, Ae. albopictus). Uzročnik je virus porodice Flaviviridae, roda Flavivirus. Postoje četiri različita serotipa virusa dengue (1-4) koji se mogu održavati u endemskim područjima svijeta. Bolest se većinom pojavljuje u tropskim i suptropskim krajevima između 35° sjeverne i 35° južne geografske širine. Infekcija može biti asimptomatska ili se očitovati kao nespecifična febrilna bolest, dengue groznica, dengue hemoragijska groznica te dengue šok sindrom. Prisutnost komarca Ae. albopictus dokazana je i u Hrvatskoj. Prvi nalaz ovog komarca zabilježen je na području Zagreba u listopadu 2004. godine, a u jesen 2005. godine i u brojnim mjestima duž jadranske obale. Tijekom 2007. godine u nas su dokazana dva importirana slučaja dengue groznice, nakon čega su importirani slučajevi kontinuirano bilježeni. U kolovozu 2010. godine zabilježen je prvi slučaj autohtone dengue groznice u Hrvatskoj, na poluotoku Pelješcu. Iako Hrvatska nije endemsko područje za dengu, prisutnost odgovarajućeg vektora, kao i moguća opasnost od importiranih slučajeva dengue zahtijevaju suvremenu i pravodobnu dijagnostiku ove bolesti. Dijagnostika virusa dengue najčešće se provodi detekcijom virusne RNK, detekcijom antigena te serološkom dijagnostikom (dokaz specifičnih protutijela)., Dengue is acute viral disease transmitted to humans by Aedes mosquitoes (Ae. aegypti, Ae. albopictus). Dengue virus belongs to the family Flaviviridae, genus Flavivirus. There are four dengue virus serotypes (1-4) which are maintained endemically. The disease is endemic in tropical and subtropical areas between latitudes 35° N and 35° S. Infections may be asymptomatic or may produce a wide spectrum of diseases: non-specific febrile illness, dengue fever, dengue haemorrhagic fever or dengue shock syndrome. For the first time in Croatia, Ae. albopictus was registered in Zagreb in October 2004. In autumn 2005, additional records of Ae. albopictus presence were made in many places along the Adriatic coast. During 2007, two cases of imported dengue fever were reported in Croatia, after which similar imported cases appeared continually. In August 2010, the first autochthonous case of dengue fever was recorded on the peninsula Pelješac. Though Croatia is not endemic for dengue, the existence of a corresponding vector and a latent threat by imported dengue cases demand state-of-the-art and timely diagnostics. The most commonly used methods in laboratory diagnosis of dengue infections involve detection of viral RNA, antigen detection and serologic methods (detection of antibodies).

The aim of this study was to determine the immunity to varicella-zoster virus (VZV) in Croatian pregnant and non-pregnant women of reproductive age. During 2007-2011, a total of 638 women aged 16-45 years were tested for the presence VZV IgM and IgG antibodies using commercial enzyme-linked immunosorbent assay. Samples positive for IgG antibodies with positive or equivocal IgM antibodies were tested for IgG avidity. The overall IgG seroprevalence was 84.3%. There was a significant increase in IgG seropositivity with age (OR=1.04 for one year increase in age ; 95% CI=1.01-1.08). The lowest seroprevalence rate was reported in the 16-20 age group (78.6%), and the highest was in the 41-45 age group (94.3%). There was no significant difference in seroprevalence among women residing in urban and rural areas (83.6% vs. 87.0%, OR=0.76, 95% CI=0.43-1.34). The results of this study have shown that a high proportion of Croatian childbearing-aged women (15.7%) who were referred to the laboratory for VZV serology testing are susceptible to VZV and, thus, at risk for contracting varicella during pregnancy. Serology testing of adolescent girls and adult women who do not have a documented history of varicella is encouraged with the aim of vaccinating seronegative girls and women against VZV before pregnancy. Additionally, testing of pregnant women is advised in order to identify susceptible women and vaccinate them after delivery.

Cilj rada bio je prikazati virološke i epidemiološke značajke non-polio enterovirusnih (NPEV) infekcija u desetogodišnjem razdoblju(2000.-2009.) u Republici Hrvatskoj. Tijekom navedenog razdoblja u Laboratoriju za enteroviruse Hrvatskog zavoda za javnozdravstvo obrađeno je 2754 kliničkih uzoraka prikupljenih od 1880 bolesnika s kliničkom slikom enterovirusne infekcije. Dijagnoza enterovirusne infekcije dokazana je izolacijom virusa u staničnoj kulturi, a virusi su tipizirani metodama neizravne imunofluorescencije i/ili testom neutralizacije. NPEV infekcija dokazana je u 394 od 1880 bolesnika (21%). Osobe muškog spola češće su obolijevale od osoba ženskog spola u omjeru 1,8:1. Broj oboljelih najveći je u dobnoj skupini predškolske i školske djece. Bolesnici u kojih su izolirani NPEV najčešće su imali dijagnozu seroznog meningitisa - 234/394 (59,4%). Infekcije su najčešće bile uzrokovane echovirusima (218/394; 55,3%), zatim coxsackie B (126/394; 32,0%) i coxsackie A (31/394; 7,9%) virusima, a rjeđe su dokazani echovirus 22 - parechovirus 1 (16/394; 4,1%) i enterovirus 71 (3/394; 0,8%). Coxsackie B5 i B4 virusi kontinuirano se detektiraju tijekom istraživanog razdoblja s učestalijom pojavom svakih 4-5 godina. Najčešći izolat echovirusa bio je echovirus 18 koji se detektira kontinuirano od 2000. do svoje epidemijske 2006. godine. Echovirus 6 i echovirus 30 također se izoliraju kontinuirano s vršnim vrijednostima 2002. i 2008., odnosno 2002., 2006. i 2008. godine., Aim: The aim of the present study was to show the virologic and epidemiological characteristics of non-polio enterovirus (NPEV) infection over a ten-year period in Croatia. Methods: During the 2000-2009 period, the Laboratory for Enteroviruses, Croatian Public Health Institute analyzed 2754 clinical samples collected from 1880 patients with a clinical picture of enteroviral infection. The diagnosis of enteroviral infection was confirmed by virus isolation in cell culture. Viruses were typed using indirect immunofluorescence and/or neutralization assay. Results: NPEV was proven in 394 (21%) of 1880 patients. Males were more commonly infected than females, at a ratio of 1.8:1, while the number of infected cases was highest among preschoolers and schoolchildren. Patients with isolated NPEV were most frequently diagnosed with aseptic meningitis (234/394; 59.4%). Infections were most commonly caused by echoviruses (218/394; 55.3%), followed by Coxsackie B (126/394; 32.0%) and Coxsackie A (31/394; 7.9%) viruses, rarely by echovirus 22 - parechovirus 1 (16/394; 4.1%) and enterovirus 71 (3/394; 0.8%). In most cases, echoviruses of the following serotypes were proven: 6, 9, 11, 13, 14, 18 and 30 (45/218, 21%; 14/218, 6.4%; 18/218, 8.3%; 15/218, 6.9%; 11/218, 5.0%; 55/218, 25.2% and 42/218, 19.3%), while serotypes 2, 3, 4, 5, 7, 12, 20, 21, 25 and 26 were evidenced in a minority of patients or individual cases. Coxsackie B5 was the predominant serotype among Coxsackie B viruses (50/126, 40%), while Coxsackie A9 was most common Coxsackie A virus (26/31, 84%). Coxsackie B5 and B4 viruses were continually detected during the study period and appeared more frequently every four to five years. The most common echovirus isolate was echovirus 18, detected continually between 2000 and the outbreak year of 2006. Echovirus 6 and echovirus 30 were also isolated continually with peaks in 2002 and 2008, or 2002, 2006 and 2008. Conclusion: The results of this study pointed to a specific pattern of the occurrence of certain NPEV serotypes in Croatia. The epidemic pattern (echovirus 18 and Coxsackie B1) was characterized by peaks with an elevated number of isolations in given years. Contrary to this, endemic viruses were isolated in similar counts every year (Coxsackie B3), or manifested milder epidemic peaks every few years (echoviruses 30 and 6, Coxsackie B4 and B5). Data on NPEV infections, given their serotype and specific pattern of occurrence, contribute significantly to prompt diagnostic, clinical and epidemiological response to NPEV infections.

Epilepsy is one of the most common neurological disorders, while neurocysticercosis caused by Taenia solium infection of the central nervous system currently represents the leading cause of secondary epilepsy in Central and South America, East and South Asia, and sub-Saharan Africa. As a result of increased migration from these endemic regions, neurocysticercosis and subsequent epilepsy are becoming a growing public health problem in developed countries as well. In order to determine the prevalence of T. solium infection in patients with epilepsy in Croatia, a retrospective serological study was conducted. A total of 770 serum samples were tested for the presence of T. solium IgG antibodies using a commercial qualitative enzyme immunoassay. The Western blot technique was used as a confirmatory test for the diagnosis. The overall seroprevalence rate of T. solium infection in patients with clinically proven epilepsy was 1.5%. Although the results have shown that infection with this tapeworm is rare in Croatia, this study hopes to increase awareness about the importance of preventive measures and benefits of accurate and timely diagnosis. Intervention measures for infection control are crucial, namely sanitation improvement, control of domestic pig-breeding, detailed meat inspection, detection and treatment of tapeworm carriers, hand washing and health education.

Cystic liver disease (CLD), presenting with solitary or multiple cysts in the liver, is a common diagnosis today, primarily due to the frequent application of modern radiological methods. There is a wide range of possible causes. CLD of infective origin is usually caused by an echinococcal species. During the past three decades a number of cystic echinococcosis (CE) control programmes have led to a significant decrease in the incidence of human hydatidosis in some endemic areas. The aim of this study was to determine the seroprevalence ofE. granulosusinfection in Croatian patients with CLD. A total of 540 serum samples from patients with hepatic cysts detected by imaging methods were screened for the presence ofE. granulosusIgG antibodies using semiquantitative enzyme-linked immunosorbent assay. The Western blot technique was used as a confirmatory test for the CE diagnosis. The overallE. granulosusseroprevalence rate in patients with CLD was 3.9%. There was no significant difference in seroprevalence rate between male and female patients (P = 0.541). According to age groups, there was a significant difference in seropositivity among age groups (P = 0.002). The highest seroprevalence rate was detected in the youngest age group (up to 18 years), both in males and females (20% and 13%, respectively). This study indicates that CE still represents a public health problem in Croatia. Preventive measures should be used to controlEchinococcusinfections, including avoidance of contact with infected dogs, egg-contaminated soil or plants; control and treatment of dogs with antihelmintics; hand washing, improved sanitation and health education.

Fungal keratitis represents one of the most difficult forms of microbial keratitis to diagnose and treat successfully. It is difficult to obtain correct diagnosis and topical antifungal preparations. Fungi can cause severe stromal necrosis and enter the anterior chamber by penetrating an intact Descemet membrane. The most common pathogens are filamentous fungi (Aspergillus and Fusarium spp.) and Candida albicans. The incidence of Trichophyton spp. keratitis is 5%. A 22 years old female contact lenses wearer after keratitis developed corneal melting syndrome, spontaneous perforation of the cornea and complicated cataract of the left eye. Conjunctival swab was sterile as well as first sample of corneal tissue and sample from the anterior chamber. Urgent therapeutic perforating keratoplasty (PK), was performed together with extracapsular cataract extraction and the implantation of the intraocular lens in the posterior chamber. The patient was treated with ciprofloxacin and diflucan (systemic therapy); with dexamethason and atropin (subconjunctivaly) and chlorhexidine, brolene, levofloxacin, polimyxin B, and dexamethason/neomycin (topically). Microbiology evaluation was performed once again following excisional biopsy of the intracameral portion of the lesion. The presence of Trichophyton spp. was finally confirmed. Itraconazole and garamycin were included in the systemic therapy. Corneal graft was clear for 17 days but decompensated 28 days after the PK. After two weeks microorganisms invaded the vitreous and caused endophthalmitis. Despite urgent pars plana vitrectomy patient developed endophthalmitis, lost light sensation and developed phthysis. Evisceration and the implantation of silicon prosthesis was done. Perforating keratoplasty is a method of choice in treating severe infectious keratitis unresponsive to conservative treatment but without the eradication of microorganisms it cannot restore the vision or save the eye. Trichophyton spp. may cause a severe disease of the anterior and posterior part of the eye which may finish with the lost of vision/eye. Prompt diagnosis and treatment of Trichophyton spp. keratitis are essential for a good visual outcome.

The meropenem yearly Susceptibility Test Information Collection (MYSTIC) programme is a global, longitudinal resistance surveillance network that monitors the activity of meropenem and compares its activity with other broadspectrum antimicrobial agents. We now report the antimicrobial efficacy of meropenem compared to other broad-spectrum agents within the selective Gram-negative pathogen groups from two Croatian Hospitals investigated between 2002-2007. A total of 1510 Gram-negative pathogens were tested and the minimum-inhibitory concentrations (MICs) were determined by broth microdilution method according to CLSI.There was no resistance to either imipenem or meropenem observed for Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis in both medical centers. High resistance rates of K. pneumoniae to ceftazidime (18%), cefepime (17%) and gentamicin (39%) are raising concern. Acinetobacter baumannii turned out to be the most resistant Gram-negative bacteria with 81% resistant to ceftazidime, 73% to cefepime, 69% to gentamicin and 71% to ciprofloxacin. Almost 20% of Pseudomonas aeruginosa strains were resistant to imipenem, 13% to meropenem, 69% to gentamicin and 38% to ciprofloxacin.The prevalence of extended-spectrum beta-lactamases (ESBLs) in E. coli was 10% and in K. pneumoniae 49%. PCR and sequencing of the amplicons revealed the presence of SHV-5 in nine E. coli strains and additional tem-1 beta-lactamase five strains. Five K. pneumoniae strains were positive for bla(SHV-5 )gene. Eight ESBL positive Enterobacter spp. strains were found to produce tem and CtX-m beta-lactamases. Plasmid-mediated AmpC beta-lactamases were not found among K. pneumoniae, E. coli and Enterobacter spp. Three A. baumannii strains from Zagreb University Center were identified by multiplex PCR as OXA-58 like producers. Six A. baumannii strains from Split University Center were found to possess an ISAba1 insertion sequence upstream of bla(OXA-51 )gene. According to our results meropenem remains an appropriate antibiotic for the treatment of severe infections caused by Gram-negative bacteria. These data indicate that despite continued use of meropenem, carbapenem resistance is not increasing among species tested, except for A. Baumannii, in the two study hospitals and suggest that clinicians can still administer carbapenems as a reliable and effective choice in managing serious nosocomial infections.

Trichinellosis is a parasitic mammalian disease caused by roundworms from the Trichinella genus. It is an important zoonosis with humans becoming infected by eating raw or inadequately cooked infested meat. The disease is widespread and represents a public health problem. According to the World Health Organisation estimations, some 11 million people are infected by Trichinella annually. Trichinellosis has a broad clinical presentation, ranging from asymptomatic to fatal. Since there are no pathognomonic signs or symptoms, clinical diagnosis is difficult and the only reliable diagnostic methods are those parasitological.

Aim of our paper is to present a case of painless Acanthamoeba keratitis in a soft contact lens wearer. A 17-year-old male, highly myopic, prolonged soft contact lens wearer, presented to us with painless red watery right eye having remarkably diminished vision. Last six weeks he was treated elsewhere for the microbial keratitis with no improvement. No pain was reported and on the direct questionnaire about it he denied it. There was marked mixed conjunctival and ciliary injection. A central stromal opacity with a pronounced surrounding corneal ring of inflammatory infiltration and epithelial defect was seen on biomicroscopy of the right eye. Circular pannus was already formed reaching epithelial defect overlying corneal ring infiltrate. Acanthamoeba spp in the corneal sample was confirmed. Prolonged therapy with 0.02% chlorhexidine digluconate solution combined with 0.1% hexamidine solution resulted in corneal healing left with a large central dense stromal opacity with circular pannus reaching peripheral third of the cornea but with very thin blood vessels and the best corrected visual acuity of 0.1 tested on Snellen chart. In conclusion, even in a lack of typical symptom for Acanthamoeba keratitis such as pain, this amoeba should be ruled out especially in a soft contact lens wearer.

A total of 142 serum specimens from Croatian children aged 3–18 years, with peripheral eosinophilia and without any clinical symptoms, were tested for the presence of Toxocara canis IgG antibodies using an enzyme-linked immunosorbent assay (ELISA) and Western blot method. The overall T. canis seropositivity rate in the samples tested was 31%. There was no statistical difference in the seroprevalence rate between boys (32.1%) and girls (29.7%, χ2 = 0.092, P = 0.761). The T. canis seropositivity varied significantly between age groups both in boys (Fisher's test P P = 0.007). The highest seroprevalence rate was found in boys aged 11–14 (42.9%) and girls aged 7–10 (44.2%). Our results showed a high seroprevalence rate of T. canis infection among children with eosinophilia. It is important that testing for toxocariasis should be included in a differential diagnosis of eosinophilia, especially in children.

Background and Aim: Postantibiotic effect (PAE) is a delay of bacterial growth after short exposure to antibiotics. The phenomenon of continuing suppression of bacterial growth after removal of β-lactamase inhibitors is termed post-β-lactamase inhibitor effect (PLIE). Recently, Pseudomonas aeruginosa strains producing metallo-β-lactamases were described in many countries of the world. The aim of the study was to investigate the PLIE of carbapenems in combinations with EDTA against VIM-MBL-positive strains of P. aeruginosa. Methods: The experiments were performed on two Pseudomonas aeruginosa isolates, one producing VIM-1 and the other producing VIM-2 metallo-β-lactamase. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBC) of imipenem and meropenem alone and combined with EDTA, time-kill curves, PAE and PLIE were performed as described previously. Results: The duration of PAE with meropenem combined with EDTA at 8 × MIC was longer against both VIM-1 and VIM-2 producer than that of imipenem with EDTA on VIM-1- and VIM-2-positive strains. The duration of PLIE was similar on both strains of P. aeruginosa regardless of the sort of carbapenem. At lower concentrations, meropenem with EDTA induced slightly longer PAE and PLIE than imipenem with EDTA. Conclusions: This study has shown that EDTA combined with carbapenems produced a significant PLIE on VIM-MBL-positive P. aeruginosa strains. The results do not have any clinical relevance so far since metal chelators such as EDTA are not used as therapeutic agents due to their toxicity.

Milker's nodule (noduli mulgentium) is a benign viral skin disease caused by parapoxvirus, a poxvirus that is endemic in cattle. The virus is usually transmitted to cattle handlers from infected cows. We present a case of a 25-year-old housewife who milked cows on her family farm and developed itchy, purplish red nodules on her fingers and hands, and lymphangiitis of the right arm. Two weeks before, several cows on her family farm were treated by a veterinarian under the diagnosis of pseudocowpox. She was treated with orally administered amoxicillin with clavulanic acid 2 g for 10 days, with complete resolution of lymphangiitis. The nodules resolved in several weeks.

SAŽETAK Slobodnoživuće amebe iz rodova Naegleria, Acanthamoeba, Balamuthia i Sappinia, široj su medicinskoj struci nepoznate, ili slabo poznate. Te amebe mogu biti uzročnici životno ugrožavajućih bolesti koje nepravodobno prepoznate i neodgovarajuće liječene, gotovo redovito završavaju ili s teškim posljedicama, ili smrću. U radu je u kratkim crtama prikazana morfologija i epidemiologija slobodnoživućih ameba, kliničke osobitosti, te mogućnosti dijagnostike, liječenja i prevencije infekcija uzrokovanih tim amebama., SUMMARY Free-living amoebas, members of the genera Naegleria, Acanthamoea, Balamuthia and Sappinia are generally unknown or little known in the medical profession. They can cause life-treathening diseases. If not recognized early enough or inapropriately treated they can cause death or serious consequences. In this paper we presented morphology, epidemiology, clinical characteristics, diagnosis, therapy and prevention of these infections.

Microorganisms in soil represent small but very active part of soil organic matter and are the driving force of nutrient supply in soils. Several studies have demonstrated that intensive tillage has notable effects on the microbial communities’ properties, causing decreases in biodiversity and in microbial biomass. Stimulation and reduction of one part of the microbial populations may induce loss of numerous important soils functions, e.g. mineralization of soil organic compounds or reduction of N20 to greenhouse neutral N2. Therefore, the aim of our study was to investigate the dynamic of microbial communities in soil planted with soybean under different tillage systems. Moreover, abundance of the nitrous- oxide reductase gene (nosZ) as an indicator of soil quality was studied. The research was established at chernozem soil of Northern Baranja region in Republic Croatia as completely randomized block design of four replicate plots for each tillage system in three years experiment. The soil was managed as followed: CT- conventional tillage (moldboard ploughing at 25-30 cm depth), DH- multiple disks harrowing (10-15 cm depth), and NT- no- tillage system. Soil samples were collected from plots in spring, summer and autumn in year 2002, 2003 and 2004. The abundance and diversity of soil microbial communities was investigated by isolating and culturing the microbes on selective media and by culture independent methods. Results of our investigation suggest that no-tillage have positive effect on communities’ structure and function and detailed results will be presented.