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Journal of IMAB - Annual Proceeding (Scientific Papers)
Publisher: Peytchinski, Gospodin Iliev
ISSN: 1312 773X (Online)
DOI: 10.5272/jimab.161cabst2010
Subject Collection: Abstracts Medicine
Date of close: May 29, 2010


Prolonged Suppressive Antimicrobial Chemotherapy for Knee and Hip Prosthetic Joint Infections: a Decade of Experience in a Bone and Joint Infections Outpatient Clinics
Athanasia S., Nikou P., Papadopoulos A. , Drakou A., Galani L., Gourgoulis GM. , Kanellaki S. , Koratzanis E. , Koukos G. , Panagopoulos P. , Paramythiotou E. , Plachouras D. , Sakka V. , Sympardi S. , Fragou A. , Giamarellou E. , Kanellakopoulou K. , Giannitsioti E.
ATTIKON University General Hospital, Athens, Greece
1) The epidemiological report and evaluation of chronic suppressive antimicrobial chemotherapy for arthroplasty infections. 2) Prognostic factors of the therapeutic outcome.
Patient-methods: Patients that have being monitored at the bone and joint infections outpatient clinic (1999-2009) and had chronic knee and hip arthroplasty infections without any further feasible surgical application. Demographics, previous surgical debridement, time elapsed from infection onset to appropriate treatment administration, clinical picture, imaging techniques and bacteriological data were recorded. Antimicrobial chemotherapy (regimen, dose and duration) were also assessed for every patient included in the study. Descriptive statistical analysis was performed, as well as univariate analysis of prognostic factors of successful outcome.
Results: Fifty patients (37 females, 13 males, mean age 68.38, range 26-98), with comorbidities (n=25), who had chronic knee (n=34) or hip (n=16) arthroplasty infection, were analyzed. Diagnosis was microbiologically documented in 34 patients. Sixteen patients were given empirical antimicrobial chemotherapy (in 80% of cases ciprofloxacin plus rifampicin). Bacteria isolated were Staphylococci (85%) and Gram negative bacteria (15%). In 60% of cases surgical debridement or revision or removal of the arthroplasty was preceded. Nineteen patients had a sinus tract, so they have been removed from the prognostic factors infection result analysis, as the emergence of a sinus tract prejudges the failure of the antimicrobial chemotherapy. Age >60 y, gender, comorbidities, empirical versus targeted antimicrobial chemotherapy, adversely affected therapeutic outcome. Patients who had received antibiotics less than 12 months for suppression, presented more often relapse of the infection compared to those being treated over. Overall, median treatment duration was 12 months, (range 3-38). Mean time of follow-up was 12 months.
Conclusion: Chronic suppressive antimicrobial chemotherapy should be administered only when all surgical procedures have failed. No attempt of suppression is effective in the presence of a sinus tract. Suppression treatment should not be given for less than 12 months in order to achieve arrest of the arthroplasty infection.

Improved detection of the fastidious sexually transmitted microorganisms Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium by mPCR.
V. Ouzounova-Raykova*, D. Yordanov, I. Mitov
Medical University-Sofia, Department of Medical Microbiology
Objectives. The aim of this study was to evaluate the prevalence of fastidious bacterial genital infections in clinical samples from healthy and symptomatic men and women applying the first in Bulgaria multiplex PCR for detection of Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium.
Methods. Totally 413 samples were included in the study. The urethral/cervical swabs were cultivated on McCoy cell line for the isolation of C. trachomatis and its detection by immunofluorescence. All the samples were subjected to DNA extraction and the DNA was used for the detection of the genome of the mentioned microorganisms by mono-PCR and afterwards by mPCR.
Results. C. trachomatis was found in 3 clinical samples (0,73%) by applying mono PCR, U. urealyticum - in 54 (13,08%), M. hominis - in 11 (2,66%), and M. genitalium - in 3 (0,73%). The results of the mPCR were identical. The amplification methods gave the same results as the cultivation and the immunofluorescence.
Conclusions. Very low is the prevalence of the genital chlamydial infection among the studied samples - 0,73%. Relatively high is the number of U. urealyticum positive probes. In all the 11 cases with detected M. hominis there was coinfection with U. urealyticum. Low is the prevalence of M. genitalium infections. Despite some statements that Mycoplasms take part in the normal flora of the genital tract we think that they may be serious pathogens. The reason for this is the fact that we found them mostly in patients with complaints and with diagnoses such as vaginitis, cervicitis, uretritis, BV, etc.

Acute Febrile Neutrophilic Dermatosis (Sweet Syndrome)
S. Pavlov*, S. Ratheva, M. Slavova, M. Nenova, M. Gospodinova, H. Varbanov, V. Gerov
Department of Dermatology and Venereology, Department of Infectios diseases, Department of Haematology, Medical University of Varna, Bulgaria
Acute febrile neutrophilic dermatosis (Sweet syndrome) is a rare disease. It was described by Robert Sweet in 1964 and only a few hundred cases were published in medical literature. Frequent association with malignancy, leukemia, multiple infections, Crohn disease and etc. were assessed. We present male patient, aged 24, who developed Sweet syndrome in the follow-up period of six months and was consecutively admitted in the clinics of Infectious diseases, Hematology and Dermatovenereology.

Pemphigus Vulgaris in Nine Years Old Child with Lambliasis
S. Pavlov*, M. Slavova, S. Racheva, P. Drumeva
Department of Dermatology and Venereology, Medical University of Varna, Bulgaria
Pemphigus is an autoimmune blistering disease that affects the age between 40 and 60 years. In children the disease is rare, here are only about a hundred cases have been reported till now. In the current case report we describe the clinical manifestations of pemphigus vulgaris in 9 years old boy with laboratory proven lambliasis.

Microbiological characterisation of Haemophilus influenzae strains isolated from patients with invasive and respiratory diseases
Kostyanev T.*, L. Setchanova, V. Ouzounova-Raykova, Y. Proevska, I. Mitov
Chair of Medical Microbiology, Faculty of Medicine, MU-Sofia, Bulgaria
Go to the full publicaton of the article in JofIMAB 2010 16(3):66-69
Objectives: To determine the prevalence of different serotypes and beta-lactamase production of invasive and respiratory H. influenzae strains isolated from children and adults before the introduction of Hib conjugate vaccine in the National Vaccination Plan.
Methods: A total of 175 H. influenzae strains were collected between 1994 and 2009 from all aged patient groups: 53 cerebrospinal fluid (CSF), 8 blood, 60 sputum, 8 ear fluid, and 46 upper respiratory tract (URT) samples. The strains were isolated from patients with invasive and community-acquired respiratory tract infections. All strains were identified according to standard microbiological methods. Serotyping was done by a coagglutination test and by molecular PCR capsular genotyping. Beta-lactamase production was determined by the chromogenic cephalosporin test with nitrocephin as substrate.
Results: Most of the isolated H. influenzae strains were from children under 5 years of age (57.7%). Overall, 61 strains belonged to serotype b (34.9%) by the means of PCR capsular typing, 1 strain was type f, and 113 isolates (64.6%) were non-typeable (non-encapsulated) H. influenzae. Among the infants and children with meningitis or other invasive infections, aged 2 month to 5 years, all strains, except one, were serotype b. In respiratory tract infections (pneumonia, otitis media, sinusitis and people with chronic pulmonary diseases - exacerbations of COPD, bronchiectasis, cystic fibrosis) the most common - 96.5% were non-typeable strains in both groups children and adults. Overall, the prevalence of beta-lactamase production was 19.4%. But, it was much higher for invasive strains from CSF isolates - 37.7%, 25% in blood samples, and 37.5% in otitis media causative strains. Beta-lactamase production was less frequent in respiratory tract isolates - in sputum 13.3% and in URT samples - 2.3%. The rate of beta-lactamase production in CSF isolates has not changed for the last 10 years.
Conclusions: PCR capsular genotyping method has to be performed for all non-b-type strains. The implementation of Hib vaccine in our country will be accompanied by a reduction in invasive diseases caused by H. influenzae type b in children, but it is not useful in preventing infections caused by non-typeable H. influenzae strains.

G. Kyuchukov, I. Kiryakov, P. Trendafilov, R. Nedelko
Saint Anna Hospital, Clinic of Neurosurgery, Varna, Bulgaria
Pyogenic vertebral osteomyelitis is of special interest to neurosurgeons because it often results in acute neurological deterioration and requires a combination of adequate surgical and conservative treatment. A group of 10 patients with the clinical and radiological signs of acute pyogenic spondylodiscitis received surgery in our clinic for the period 2007-2009. The age ranged from 50- 70 years. All of them had underlying diseases such as diabetes mellitus or pneumonia.  All the infections were localized in the thoraco- lumbar  ( Th10- L2) region and the main isolated organism was Staphylococcus aureus. A delay in diagnosis was registered in all cases. The first and the main symptom was pain and treatment with NSAIDs have been prescribed till the development of neurological deficits. That’s why the patients were admitted in our clinic with rapidly progressing neurological deficit, fever, bowel and bladder dysfunction. They underwent immediate surgery- including decompression and stabilization. An empiric antibiotic therapy was started and necessary corrections were made after receiving the antibiogram. It’s well known that the intervertebral disc doesn’t have its own blood supply. In all of our cases osteolytic  destruction of both vertebral discal surfaces were seen, and discitis seems to be secondarly involved in this infection. So the functional unit of the vertebral column fails due to deformation in saggital and axial plains mainly, compressing the neural structures and  resulting in neurological deterioration. Decompression of neural elements and restoration of the vertebral column biomechanics were the aim of surgery. A posterior approach was used in our cases. Pain relief was registered immediately the day after surgery. Nonspecific spontaneous spinal epidural abscesses and spondylodiscitis are rare clinical conditions. Their incidence, however, appears to be on the rise, which may be attributed to various factors such as impaired immunocompetence. Because acute pyogenic vertebral osteomyelitis often results in acute neurological deterioration and requires a combination of adequate surgical and conservative treatment, it is of special importance to neurosurgeons.

Immunity and Infections - Does Gender Matter?
V. Sarafian
Department of Biology, Medical University-Plovdiv, Bulgaria
Gender-specific medicine studies the biological and physiological differences between the human sexes and how that affects differences in disease. Recently medical research has started to understand the importance of taking the sex into count as the symptoms and responses to medical treatment may be very different between sexes.
Disease frequency, predisposition, progression and even therapeutic approach are gender dependent. Immune response shows specific differences in men and women, which are mostly associated with hormonal effects. Genetic and epigenetic factors also modulate disease development and the intensity of immune reactions.
Immune-neuro-endocrine regulation is essential for the precise functioning of the immune system. Glucocorticoids, estrogens, progestins and androgens influence immune cells via specific receptor-mediated interactions. Gender differences include the domination of Th1 immune response in males and Th2 in females, but also a number of sex related characteristics in cytokine secretion, antibody production and cell activation.
Complex hormone-induced molecular mechanisms determine the prevalence of definite pathology in both sexes. Autoimmune diseases, hypersensitivity reactions, infectious diseases, traumatic conditions and many others show gender-specific frequency, clinical course and prognosis. Ageing and longevity are also gender dependent.
Bacterial, protozoal and viral infections trigger different immune response mechanisms in males and females. Gender should be taken into account in vaccine development and evaluation because of specific immunodominant epitopes eliciting different immune response to vaccination in both sexes.
The obscure pathogenetic mechanisms of a number of diseases could be elucidated through the precise knowledge of immuno-endocrine interactions. Undoubtedly, novel possibilities for adequate prevention, diagnostics and treatment will appear soon.

Approaches for Treatment of Early Lyme Borreliosis
Iva Christova
National Center of Infectious and Parasitic Diseases, Sofia,  Bulgaria
Lyme borreliosis is the most commonly reported tick-borne disease in the Northern hemisphere. The disease affects mainly skin, nervous system, joints, and heart. Three separate stages in the clinical development of the disease could be distinguished – early localized, early disseminated, and late Lyme borreliosis. Except erythema migrans (EM), the clinical hallmark of early localized Lyme borreliosis, disease expressions are not specific and need laboratory confirmation. Clinical effectiveness of antibiotic treatment decreases markedly with the progression of the disease. Therefore, early diagnostics and treatment are of great importance. Standard antibiotic treatment of early Lyme borreliosis includes treatment with doxycycline. In vitro studies have shown antiborrelial activity of azithromycin and amoxicillin. Clinical trials in Europe and the USA have revealed controversial results for their efficacy in the treatment of early Lyme borreliosis. In this study, we compared clinical outcome in patients with erythema migrans treated with doxycycline and with azithromycin or ampicillin. We established the applicability of all three antibiotics to Bulgarian patients with early Lyme borreliosis.

Modern Internet Based Instruments for Monitoring and Prediction of the Therapy Success of HIV-1 Infected Patients
Ivailo Alexiev,* D. Beshkov, V. Georgieva, L. Karamacheva
National HIV Confirmatory Lab, NCIPD, Sofia, Bulgaria
Objectives: In the recent years a lot of Internet based software tools were developed in the field of HIV-1 infection monitoring. The complexity of HIV-1 infection and the expanding role of laboratory examinations including multidisciplinary specialists impose the necessity of developing new powerful instruments for analysis and interpretation of the data from molecular biology methods: viral load, CD cells and particularly resistance tests. The expert's knowledge and local software together with the Internet based tools for monitoring and prediction of HIV-1 infection become an important part of patients care.
Methods: Currently we use Internet based software tools for two different tasks: resistance interpretation of the viral genome and Subtyping HIV. The resistance interpretation tools can be divided in two groups, HIVDB - located in Stanford, California, USA, the gold standard in the field and European tools: ANRS - France, GRADE – Germany, Geno2pheno - Germany, REGA - Belgium, and recently established EURESIST colaborative European progect located in Sweden Subtyping Internet based software tools are REGA, HIVDB, NCBI - USA, Los Alamos and recently established COMET Subtyping tool
Results: We use Internet based software tools for resistance interpretation of the viral genome to support our knowledge especially in multi drug experienced patients with many cross resistant mutations. For subtyping purposes it is useful to make preliminary analysis with Internet accessible tools prior manual phylogenetic analysis. The results from automatic subtyping are similar to manual methods and the main advantage is high speed of calculations due to relatively simplified algorithms in the process.
Conclusion: The extreme inclination for mutations pursuing HIV-1 replication and as a consequence resistance to antiretroviral therapy enforces frequent testing for resistance by genotyping viral genome. Data of the sequences obtained by sequencing of partial viral genome undergo software analysis for mutation scoring. Newly developed algorithms can go further and make complex analysis by combining a set of data including sequences, CD4 cell count, viral load and treatment regimen giving current and even predictive status of HIV-1 infected patient.
These tools have been used for many years from the experts all over the world and become inseparable part of clinical management. Consensus mutations in the data sets and interpretation algorithms are proven to be one of the most relevant to disease management. Due to the constant update with new data and interpretation algorithm improvement we found resistance and subtype Internet based systems as a useful support in addition to local software regarding resistance and manual methods for Subtyping HIV-1.

Hadjieva N 1, Petrov M M 2, Velinova V 1
1. University Hospital “Queen Giovanna – ISUL”; 2. National Centre of Infectious and Parasitic Diseases, Sofia, Bulgaria
University Hospital “Queen Giovanna – ISUL” is one of the leading Bulgarian hospitals with more than 75 years of history and more then 550 beds that serve between 12000 and 25000 patients annually. The following paper presents a 15 year 1994 - 2008) activities in the Clinical Microbiology and Virology Laboratory in “Queen Giovanna – ISUL”. Some tendencies in microbiological structure of infections in hospital are presented, frequency of bacterial/fungal isolation and some susceptibility patterns of certain bacteria/antibiotic pairs for the selected period of time are presented.
There was a two and half fold reduction in the number of samples from 15000 for 1994 to almost 6000 for 2008, based on the lack of clinical standards for "Infection/Hospital infection" and on outdated as a structure and function Standard Operative Procedures named Clinical Paths in Bulgaria.
Gram-negative bacteria prevailed as isolates in our hospital till 2001 when Gram-positive bacteria outnumbered them. Unexpectedly, in 2008 Gram-negatives gained force again. There are no significant differences between Bulgaria and the other European countries in the structure/aetiology of infections, evidenced from that the most common pathogen for 2008 is E. coli (30%), followed by CNS (22%) and Staphylococcus aureus (16%).
The observed levels of resistance give us a place in the "golden middle" among the family of European countries. There are still cases of overuse and misuse of antibiotics in the University Hospital “Queen Giovanna – ISUL”, despite the fact that this is the first Bulgarian hospital with real implementation of hospital antibiotic policy since 1993. A special policy for the minimizing the risk of multidrugresistant Gram negative infections should be implemented by the Infection Control Team in the University Hospital - Queen Giovanna - ISUL.

Surgical Personnel’ Adherence to Antibiotic Policies
E. Keuleyan*, G. Kirov, M. Kondarev, I. Lozev, D. Vezeva, S. Toujarov, N. Smilov
Medical Institute – Ministry of the Interior, Sofia, Bulgaria
This work aimed at studying the adherence of personnel of the Surgery clinic to antibiotic policies in place.
Methods. Antimicrobial resistance surveillance of the alert resistant microorganisms (Staphylococcus aureus (MRSA), Enterobacteriaceae (ESBL-producing), Pseudomonas aeruginosa, MDR); Antibiotic consumption calculation (ABC calc, D. Monnet); Audits of antibiotic prescriptions, and Inquiries – were performed.
Results. Current antibiotic policies in the Surgery clinic (50-bed) were prepared, discussed and introduced in 2003. Since then, five 3-month audits of antibiotic prescriptions took place. During the period 2003-2009, problem resistant organisms were: MRSA, 11-36 %; ESBL-producing Enterobacteriaceae, 14-23 %; MDR P. aeruginosa, 30-47 % and carbapenem-resistant, 0-35 %. Antibiotic consumption was between 47.9 and 61.9 DDD/100 bed-day, and first generation cephalosporins were the top used antibiotics (29.5-35.2 DDD/100 bed-day). Two inquiries (2007 and 2009) revealed: - a good compliance with the policies, with the exception of the length of antibiotic prophylaxis, - and a comprehensive knowledge on antibiotics and antibiotic resistance.
Conclusion. With one exception (the length of antibiotic prophylaxis) this study revealed good adherence to antibiotic policies, as well as professional attitudes towards the rational use of antibiotics.

Acute Viral Torticollis: Case Report
Ivanov, B., Dimitrov, I.*, Deleva, N.
Neurology Department, Medical University, Varna, Bulgaria
Objectives: Torticollis may be an alarming symptom of a life threatening condition. Its evaluation imposes consideration of traumatic, inflammatory, neoplastic, systemic or neurodegenerative underlying pathology.
Methods: Case report
Results: We report a case of acute torticollis in the course of viral rhinopharyngitis in a 24-year-old female. The viral infection resolved within a week, but torticollis persisted and the patient presented at the neurology department. Blood tests, consultations and imaging diagnostics were all normal. After 10 days of treatment with clonazepam the symptoms resolved completely.
Conclusion: Acute viral torticollis is typical in childhood, especially between 2 and 4 years of age. Meanwhile its incidence in adults is increasing. The patient we report had a benign disease course, symptoms resolved over time with drug treatment. In spite of all, attention must be directed to such differentials as meningitis, retropharyngeal abscesses, tuberculosis or pneumonia. Torticollis can be a consequence of a traumatic injury or drug overdose. All that implies a thorough history and examination in order to rule out potentially dangerous causes.

The serum concentration of 25-OH vitamin D in various infections
S. Iacob 1, D. Banica 2, E. Panaitescu 1, M. Cojocaru 3 *, D. Iacob 1
1)“Carol Davila” University, Bucharest, 2)“Marius Nasta” Institute of Pneumology Bucharest, 3) Titu Maiorescu University Bucharest, Romania
Recent studies suggested the immunomodulatory role of vitamin D in various infections. We evaluated the plasma level of 25-hydroxyvitamin D [25(OH) D] in different types of infections and its correlation with CD4 T helper lymphocytes /CD8 cytotoxic-suppressive lymphocytes ratio as a measure of the immune system status.
The study was performed on 68 patients, aged 17-73 years hospitalized in Matei Bals Institute, Bucharest, Romania, out of which 22 were diagnosed with viral infections, 16 with bacterial infections and 30 healthy controls. All patients had a favorable evolution. Circulating 25(OH) D level was measured using ELISA (IDS LTD).The level of 25(OH) D was defined as normal (> 80 nmol/L), insufficient (50-80nmol/L) or deficient (<50 nmol/L). The CD4 and CD8 lymphocytes count was determined using flow cytometry; the normal CD4/CD8 ratio was defined as >1.5
The mean serum concentration of 25(OH) D in bacterial infections was of 32.42 (+/-11.50) nmol/L and slightly lower in viral infections 31.31 (+/-7) nmolL.By comparison, the mean value in healthy controls was 28.62 (+/-8.3).The CD4/CD8 ratio was 1.4 (+/-0.7) in bacterial infections, 1.9 (+/- 0.7) in viral infections and 1.5 (+/- 0.8) in controls. The linear Pearson correlation between the level of 25(OH) D and the CD4/CD8 ratio was weakly positive in healthy controls and held no significance related to the studied infections.
Conclusion: All subjects presented a marked deficiency of 25(OH) D. No correlation was found between the serum values of 25(OH) D and the total number or the ratio of CD4/CD8 lymphocytes.The extremely low level of 25(OH) D had no negative consequence on the clinical outcome or the immunologic response of the evaluated subjects.
This work was supported by CNCSIS - UEFISCSU, PNII-Idei, cod 1165/2008

Plasma Levels of Cathelicidin LL37 Antimicrobial Peptide in Various Infections
S. Iacob 1, D. Banica 2, E. Panaitescu 1, M. Cojocaru 3 *, D. Iacob 1
1)“Carol Davila” University, Bucharest, 2)“Marius Nasta” Institute of Pneumology Bucharest, 3) Titu Maiorescu University Bucharest, Romania
Objective. Cathelicidin-LL37 (LL37) is a human antimicrobial peptide with wide antibacterial and antiviral spectrum. Its transcription is promoted by vitamin D in several cells and tissues, but mostly in neutrophils. Although recent evidence suggests that LL37 plasma level increases in some cutaneous infections and sepsis, little data is available on this subject. Our objective was to assess the functional significance of LL37 plasma level in various infections.
Methods: The study was carried on 52 patients (19 men, 33 women), aged 17-83 years, hospitalized in Matei Bals Institute, Bucharest, Romania. 25 of these were diagnosed with viral infections (19 with HIV infection, 4 with varicella zoster virus infection, 2 with human papilloma virus infection), 6 patients with mixed viral-bacterial pulmonary infections, 7 patients with bacterial infections (3 streptococcal infections, 4 staphylococcal infections) and 14 healthy controls.
LL37 serum concentrations were measured using ELISA method (Hycult biotech).
Results: LL37 mean plasma level in patients with viral infections was 9.353(+/- 6.164) ng/ml, higher in bacterial infections 10.727 (+/- 7.558) ng/ml and in HIV infected patients 11.222 (+/- 8.262) ng/ml. The mean value in the control group was 9.307 (+/-5.915) ng/ml. All patients presented a marked deficiency of vitamin D. (25OH vitamin D level <47nmol/L). Weak correlations were founded between LL37 and neutrophils number (R=0.583) and also between LL37 and the total number of leukocytes (R=0.598).
Conclusion: LL37 plasma levels in the aforementioned infections were very low compared with other studies but not significantly different from healthy subjects. We presume that a higher level and thus the microbicidal effect of LL37 is primarily exerted in tissues rather than in plasma where it is rapidly inactivated.
This work was supported by CNCSIS - UEFISCSU, PNII-Idei, cod 1165/2008

The Dynamics of Antimicrobial Resistance of Salmonella typhimurium Isolates.
*Naxhije Hila.* *Ariola Devolli. **Klementina Puto. *Sotir Mali. **Zheni Brahimaj. *Erleta Peqini. **Aida Dervishi.
*Departament of Biology, “A. Xhuvani” Universitety, Elbasan, Albania. **Departament of Biotecnology, University of Tirana, Albania
Salmonella typhimurium is the most common pathogen isolated in foodstuffs toxin infections. The development of antibiotic resistance in Salmonella typhimurium over the last twenty years is caused by an extensive application of some antibiotics. The study of antimicrobial resistance of Salmonella typhimurium isolates helps the physicians in using the indispensable antibiotic and replacing the resistant antibiotics with new ones.
The aim of this study was to determine the prevalence of antimicrobial resistance of Salmonella typhimurium isolates of persons with foodstuff’s toxin infections in Elbasan region during the years 1985-2004, to which are made the antibiograms and the antimicrobial resistance. This study includes 2931 Salmonella typhimurium strains in this period of time. All the strains were tested using disk diffusion method (with antibiotics: AM, S, TE, C, K, GM, SXT, NA and CIP). Salmonella typhimurium strains have been more resistant to Ampicilin (98.29 %), Tetracycline (93.48 %, Streptomycin (78.92 %), Bactrim (64.14 %).
The reason of passivity of such antibiotics against Salmonella typhimurium is their use, in most cases, without doing the antibiograms in each case and physicians have used very often the above antibiotics, bringing about the intensification of bacteria’s resistance towards them.

Effect of microbes on immune regulation and development of autoimmune and  immune-mediated diseases
Manole Cojocaru1, Inimioara Mihaela Cojocaru2, Simona Alexandra Iacob3, Gabriela Bancescu4, Manuela Radu-Popescu4, Lucica Rosu5, Isabela Silosi5,
1”Titu Maiorescu” University, Faculty of Medicine, Department of Physiology,  ”Dr I. Stoia” Centre for Rheumatic Diseases, Bucharest, 2”Carol Davila” University of Medicine and Pharmacy, Colentina Clinical Hospital, Department of Neurology, Bucharest, 3”Carol Davila” University of Medicine and Pharmacy, ”Matei Bals” Institute of Infectious Diseases, Bucharest,4”Carol Davila” University of Medicine and Pharmacy, Department of Bacteriology, Bucharest, 5”Carol Davila” University of Medicine and Pharmacy, Department of Immunology, Craiova, Romania
Infectious agents and/or their products have been implicated in the pathogenesis of autoimmune and chronic inflammatory diseases. Various mechanisms by which pathogens could induce autoimmune or immune-mediated diseases have been suggested. There is evidence that the development of certain autoimmune diseases may be associated with a bacterial or viral infection that stimulates production of antibodies and immune cells called T cells, which are targeted against bacterial proteins that closely resemble “self” proteins, leading to cross-reactivity with healthy tissues. Microbial antigens have the potential to initiate auto reactivity through molecular mimicry, polyclonal activation, or the release of previously sequestered antigens. Microbial infection can also cause polyclonal activation of auto reactive lymphocytes. Microbes that kill cells can cause inflammation, the release of sequestered antigens, and autoimmunity.
It is clear that inflammation, even in the absence of infection, can trigger polyclonal activation and auto reactivity. The organism may directly generate an immune response by its continued presence. Alternately, the organism may induce an immune response, possibly by revealing self antigens that are normally sequestered from the immune system, and this auto reactive response then becomes self-sustaining.
A role for super antigens, which can be of viral or bacterial origin, has also been postulated. Infection could cause the initial activation of the lymphocytes that mediate these diseases and auto antigen could sustain the activation that persists even after the eradication of the infectious agent. Many of the immune diseases associated with infection have a genetic component, suggesting that genetic susceptibility may play a role in the development of pathologic immune response to microorganisms.
Investigation of the role of pathogens in the development and regulation of the immune response in autoimmune or chronic immune-mediated inflammatory diseases may lead to new preventive or therapeutic strategies for these diseases.

Antibiotic stewardship: characteristics, principles, objectives
F.  Allerberger
Medical University Vienna,  Austria
Prevention and control of antimicrobial resistance relies on two complementary strategies: 1. Infection control measures to control spread of multi-drug resistant organisms, and 2. Optimisation of antibiotic usage for therapy and prophylaxis. The latter endeavour is commonly summarized under the term antibiotic stewardship. Council recommendation of 9 June 2009 on patient safety, including the prevention and control of healthcare associated infections (2009/C 151/01) asked healthcare institutions to have in place a control programme addressing aspects such as organisational and structural arrangements, diagnostic and therapeutic procedures (for example antimicrobial stewardship). There is an urgent need for agreement on principles and key components of antibiotic stewardship to support the European Union member states in developing their national or regional programmes. A proposal for a conceptual framework was drafted during an international expert workshop on hospital antibiotic stewardship organized under the Czech presidency in Prague on April 15, 2009. This document aimed at defining structural and organisational requirements to optimize antibiotic use for hospitalized patients (Allerberger F, Gareis R, Jindruk V, Struelens MJ: Antibiotic stewardship implementation in the European Union: The way forward. Expert Rev. Anti Infect. Ther., 2009). Optimisation should aim at improving patient outcomes, ensuring cost-effective therapy and reducing adverse health and ecological effects of antimicrobial use, including drug resistance. Antibiotic stewardship, as a function and responsibility of national health authorities, of hospitals, and community care practitioners, has to be distinguished from antibiotic stewardship programmes. Antibiotic stewardship programmes are composed of organizational structures and action plans for implementing antibiotic stewardship. Whenever an antibiotic stewardship programme is implemented, a multitude of interventions are required. Single interventions, like hiring an antibiotic officer or performing antibiotic stewardship related training, are never sufficient alone. A programme is per definition a combination of several related interventions.

Microbiological diagnosis of Clostridium difficile infection
F.  Allerberger
Medical University Vienna, Austria
Disease due to C. difficile may be associated with a wide range of clinical manifestations. The diagnosis of CDI therefore is based on clinical history/presentation in combination with laboratory tests. Microbiological tests for C. difficile rely on: 1.) C. difficile products (glutamate dehydrogenase (GDH), aromatic fatty acids, toxins A or B), 2.) culture methods for the detection of toxin-producing C. difficile (toxigenic culture), and 3.) tests for C. difficile genes (PCRs for 16S RNA, toxin genes, genes for GDH). The time consuming cell culture cytotoxicity assay is considered the gold standard for the detection of C. difficile toxins. Most laboratories use rapid and easy-to-perform enzyme immunoassays detecting toxin A or B or the enzyme glutamate dehydrogenase, which is produced by both toxigenic and non-toxigenic strains. Real-time PCR is increasingly used to detect toxin genes of C. difficile directly from stool samples. ESCMID recommendations for diagnosing CDI propose a two-step protocol: reporting negative test results as negative, but re-testing positive first results with a second method [Crobach MJT, Dekkers OM, Wilcox MH, Kuijper EJ. European Society of Clinical Microbiology and Infectious Diseases (ESCMID) (2009) Data review and recommendations for diagnosing Clostridium difficile-infection (CDI). Clin Microbiol Infect 15: 1053-1066]. Samples with an initial positive test result, but a negative second test result, require testing with a reference method as a third step. Submission of multiple samples per patient for testing by one of the above listed tests will not increase the rate of detection of CDI in an endemic setting; in an epidemic setting, negative predictive values of these tests will be significantly lower due to the higher prevalence of disease. Toxigenic culture, i.e. culture followed by in vitro toxin detection of the isolated strains, should at least be performed on all initial stool specimens (yielding positive toxin results) from patients with severe CDI (i.e. infections with lethal outcome or requiring surgical intervention or intensive care) to allow for subtyping of isolates. PCR ribotyping, based on size variations of the 16S-23S intergenic spacer region, is the preferred typing method for C. difficile in Europe. Capillary gel electrophoresis-based PCR ribotyping might be a way of overcoming the problems associated with inter-laboratory comparisons of typing results (

In Vitro Activities of Quinolones Against Nosocomial Staphylococcus aureus
Kalliopi Chatzopoulou2, Asimoula Koteli1,* Theodosios Lavdas1, Konstantinos Tsepanis1, Savvato Tsingene1 , Paransem Lagoudaki2
1Haematology Laboratory, 2Microbiology Laboratory, “G.Gennimatas” General Hospital of Thessaloniki, Greece
Introduction: Nosocomial methicilin-resistant Staphylococcus aureus are know to be one of the main problems of antimicrobial therapy in term of resistance of this pathogen to all b-lactams and to many other classes of antimicrobials.
Purpose: To compare the in vitro activity of ciprofloxacin, levofloxacin and moxifloxacin against clinical isolates of Staphylococcus aureus from varius species in a general Hospital.
Material and methods: A total 3311 positive cultures between 2006-2007 were tested. 121 strains were indentified as Staphylococcus aureus from various clinical samples from patients hospitalized in our Hospital. All specimens were routinely inoculated into culture media. The blood cultures were incubated in Bactec 9120 system (Becton DickinsonВ®) and in aerobic plus, anaerobic plus and mycosis vials. Identification of microorganisms and susceptibility test was performed with the Vitek 2 system (BioMerieuxВ®France) and susceptibility disc diffusion method according to CLCI directions. Oxacillin MIC test was determined with Vitek 2 automated system (BioMerieuxВ®) and confirmed furthermore with E-test. Control of resistance to oxacillin was accomplished with oxacillin disk 1 mg on Muller Hinton agar with the addition of NaCl 4% and was confirmed with the detection of PBP2a of MRSA ( Slidex MRSA - BioMerieuxВ® France). In order to confirm this we used the nutritious material BBL- CHROMagar- MRSA of Becton Dickinson for 2006-2007. In order to examine the production of b-lactamase we used cefinase discs (cefinaseTM CEF-F) BioMerieux, France. The S. aureus ATCC 29213 was used as a control strain.
Results: Among 121 isolates tested, 52 (42.8%) were methicillin-resistance (MRSA) and 69 (57.25) methicillin-sensitive (MSSA). The resistance rates (%) were: ciprofloxacin 19, levofloxacin 12, moxifloxacin 8, erythromycin 36, rifampicin 6, gentamycin 84, clindamycin 20, tetracycline 44, linezolid 0, quinopristin/dulfopristin 1, teicoplanin 0, vancomycin 0.
Conclusion: Moxifloxacin has showed good activity both methicillin-resistance and methicillin-sensitive S. aureus followed by levofloxacin and ciprofloxacin. However, moxifloxacin and levofloxacin have a reduced activity against non-susceptibility to ciprofloxacin S. aureus isolates.

Antibiotic Resistance Rates in Pediatric Infections by Enterococci
Asimoula Koteli1,* Theodosios Lavdas1, Konstantinos Tsepanis1, Savvato Tsingene1 , Kalliopi Chatzopoulou2, Paransem Lagoudaki2
1Microbiology Laboratory, 2Haematology Laboratory, “G.Gennimatas” General Hospital of Thessaloniki, Greece
BACKGROUND: Enterococci are increacing in importance as nososomial pathogens. They are the 3rd most frequent cause of infection in hospitals, responsible for about 12% of hospital infections.
AIM: to determine the antimicrobial resistance patterns of enterococci isolated from clinical specimens of pediatric patients treated at our Hospital.
MATERIALS and METHODS: 86 clinical isolates of enterococci were collected for further analysis during the period January 2006 through August 2009. Identification of microorganisms and susceptibility test were performed by the Vitek 2 (BioMerieuxВ®, France). The minimum inhibitory concentrations (MICs) of five antimicrobial agents were determined by E-test method (SolnaВ®, Sweden) for each isolate. Disk diffusion tests were also performed and the results were compared with those reported by clinical laboratory. Because gentamycin susceptibility tests showed inconsistent results in many isolates, MIC determinations by the micro-broth dilution method were also performed for these isolates. All isolates were tested for ОІ-lactamase production. Enterococcus faecalis ATCC 29212 was used as a control.
RESULTS: 43 strains out of 86 strains tested were E.faecalis, 37 E.faecium, 2 E.gallinarum, 2 E.avium, 1 E.durans and 1 E. hirae. Major sources of these isolates included urine, tracheal secretions, traumas and only 5 E.faecalis from blood. The resistance rates (%) for E.faecium and E.faecalis were: ampicillin74/28, ciprofloxacin 69/10, erythromycin 89/69, gentamycin high level 59/34, streptomycin high level 70/38, teicoplanin 6/0, tetracycline 26/69, trimethoprim/sulfa 97/90, vancomycin 6/0. 6 VRE strains (5 E. faecium from urine and 1 E.faecium from blood) developed high resistance to vancomycin and teicoplanin, suggestive that they belong to the phenotype VanA.
CONCLUSIONS: E.faecium showed higher resistance as E.faecalis. The high resistance to aminoglycosides and the presence of VRE strains consist a serious problem for the treatment of infections caused by them. Increased alertness should be given to the occurrence of multi -resistant E.faecium and mechanisms causing its abundance, such as selection via antibiotic pre-treatment or ineffective empirical antibiotic treatment.

Isolation of S. lugdunensis Strains from Clinical Specimens
Asimoula Koteli1,* Kalliopi Chatzopoulou2, Konstantinos Tsepanis1, Theodosios Lavdas1, Savvato Tsingene1 , Paransem Lagoudaki2
1Microbiology Laboratory, 2Haematology Laboratory, “G. Gennimatas” General Hospital of Thessaloniki, Greece
INTRODUCTION: S.lugdunensis belongs to coagulase-negative Staphylococci (CoNS) and has been described as particularly pathogenic after the detection of virulent factors similar to those of S.aureus.
AIM: Study of Isolation frequency of S.lugdunensis strains from various clinical specimens from our hospital patients.
METHOD: We studied retrospectively 8771 cultures of various clinical specimens from the period 2003-2008. Identification was achieved by the use of catalase, coagulase, DNase test with the use of VITEK I and II (BioMerieuxВ®, France) which was also used for the antibiotic sensitivity test. Oxacillin resistance was detected with oxacillin disks (1Ојg) and Mueller Hinton agar containing 6Ојg/ml of Oxacillin.
RESULTS: 3834 of the cultures were done during 2003-2005 and another 4937 during 2006-2008. 120/174 strains of S.aureus and 599/652 CoNS were isolated respectively. No S.lugdunensis strains were isolated during 2003-2005. During 2006-2008 10 strains were isolated from different patients with a frequency of 1/4/5 per year respectively. Isolated strains distribution was as follows: 3 from I.C.U., 3 from Surgery clinic, 2 from Orthopaedic clinic , 1 from Urology clinic and 1 outpatient. 2006 strain was originated from a female, all four 2007 strains from males and in 2008 four strains originated from females and only one from a male. As far as it concerts the type of specimen, 3 strains were isolated from blood, 2 from pus, 2 from purulent wounds, 1 from central venous catheter, 1 from peritoneal fluid from a patient with pelvic abscess and 1 from prostatic fluid of a patient with prostatitis symptomatology. Three out of all strains (2 from blood and 1 from wound) were resistant to oxacillin. All strains were sensitive to vancomycin, teicoplanin, linezolid, quinopristin/dalfopristin, trimethoprim/sulfamethoxazole , clindamycin, rifampin and ciprofloxacin.

Liver Dysfunction in Murine Typhus: A Study of 161 Cases
S. Doukakis, M. Papadakis, M. Daoukakis, A. Nikolidaki, D. Triantafillidou, G. Polymili, K. Bambili, M. Junie, S. Kastanakis
Saint George General Hospital Chania, Greece
The most frequent biochemical abnormality in murine typhus is a mild elevation of AST but ALT and LDH are often elevated in parallel. A hundred and sixty-one patients with compatible clinical status of murine typhus and high serological titers of antibodies against Rickettsia typhi, were studied from our team between January 1993 and December 2007. For the study of their hepatic function three serum samples were obtained from each patient. The first sample was obtained on admission, approximately 9 days after the onset of the disease. The second sample approximately two weeks after the first. The third sample, taken from the one third of the patients, was obtained one month after the second. On admission (first sample) 139/160 patients (86.8%) presented an elevation of AST, 111/160 patients (69.4%) presented an elevation of ALT and 134/157 patients (85.3%) presented an increase of LDH. The median value of AST, ALT and LDH was 59.0 U/L, 52.0 U/L and 338.0 U/L respectively. Two weeks later (second sample) 126/150 patients (84.0%) presented an increase of AST, 112/148 patients (75.7%) presented an increase of ALT and 117/145 patients (78.0%) presented an elevation of LDH. The median value of AST, ALT and LDH was 52.0 U/L, 53.5 U/L and 333.0 U/L respectively. One month later (third sample) 14/42 patients (34.0%) presented an increase of AST, 12/42 patients (29.0%) presented an increase of ALT and 8/42 patients (19.0%) presented an elevation of LDH. The median value of AST, ALT and LDH was 28.0 U/L, 24.0 U/L and 142 U/L respectively. Our study showed that the time of normal restoration of liver function was about two months after the onset of the disease.

Savov, E*, E. Kjoseva, I. Gergova, M. Borisova, N. Vladov, N. Petrov, E. Belokonski, K. Katsarov, A. Trifonova, I. Todorova
Military Medical Academy, Sofia, Bulgaria
The infection is one of the leading causes of morbidity and mortality in liver transplant’ patients. Aerobic and facultative anaerobic Gram-negative and Gram-positive bacteria accounted the most part as a cause of bacterial infections development. The aim of this study is to determinate the structure of bacterial infections in patients after liver transplantation, to estimate the periods of time when they are likely to occur and to accept the role of microorganisms causing these type of infections. Ten patients / 8 man and 2 women / after liver transplantation in the period of time between January 30, 2008 and October 16, 2009 were reviewed. The protocols for induction of immunosuppression preparation of the recipient for liver transplantation, monitoring of the patients after liver transplantation, prophylaxis and treatment of patients after liver transplantation were approved by the Executive Agency for transplantation belonging to the Ministry of Health. From the data presented above it can be concluded that the infections / intrabdominal, wound infections, bloodstream infections /BSI/, urine infections/ occurred during the first 20 days to 4 weeks after operation and there is not discrepancy in the number between the Gram-negative and Gram-positive bacteria, isolated as a causative agents of the infections in the patients studied. Bacterial infections are common complications in patients after liver transplantation. The patients mortality is due to bacterial infection complications but not to the transplant rejection.

Assessing antibiotic resistance of nosocomial strains in multiprofile hospital
Savov, E*., I. Gergova, M. Borisova, E. Kjoseva, A. Trifonova
Department of Microbiology, Military Medical Academy / MMA /, Sofia, Bulgaria
Antibiotic resistance is a worldwide public health problem that continues to grow. Multiresistant bacteria are significant problem especially for the hospital infectious pathology and the antimicrobial resistance can result in increases morbidity, disease and economic burden and mortality. This study represents the data, obtained as a part of monitoring of the resistance of some problematic for hospital pathology bacteria - Esherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus faecalis for the last three years / 2006- 2008 / at MMA. Also, we make an attempt to recognize and explain some of the resistance-mechanisms in E.coli, K.pneumoniae and A.baumannii There was registered a significant increase in the proportion of multiresistant E.coli and K.pneumoniae to beta-lactams and probably it is connected with the production of extended-spectrum beta-lactamases /ESBL/. For E.coli the relative part of the strains, producing ESBL is about 20% and for K.pneumoniae strains this value is much more - 67.8% during 2008. There was registered high level resistance of A.baumannii strains to carbapenems - 82.9% for imipenem and 85.8% for meropenem and it was most likely associated with the production of Oxa 23 and Oxa 58 carbapemenases. The relative part of MRSA is 25%. By this way, the surveillance of antimicrobial resistant portions provides data that are needed to raise the awareness to the problem and instigate necessary optimal interventions.

Study on Seroprevalence of Mumps - Specific IgG Antibodies in a Healthy Population
M. Karcheva *, M. Atanasova1, I. Makaveev2, M. Daskalova2
*,2 Department of Epidemiology, Parasitology and Tropical Medicine, Medical University - Pleven, Bulgaria, Medical University - Pleven, Bulgaria
Go to the full publicaton of the article in JofIMAB 2010 16(3):23-26
Mumps is a vaccine preventable viral infection. Its typical clinical manifestations are characterized by pain and swelling of the salivary glands, fever, and fatigue. Often other organs are affected - testes in males after puberty (orchitis), ovaries in women (ooforitis), pancreas (pancreatitis), central nervous system (meningities). The use of specific immune prophylaxis led to a significant success in the fight against mumps, but there are still unresolved issues related to the immunological and epidemiological effectiveness of the vaccines. The disease continues to interest researchers today. The main issues being tackled are related to the conduct of virological, clinical and sero-epidemiological studies in different countries. Objectives of the study is to determine the frequency distribution of mumps-specific IgG antibodies in healthy populations in the region of Pleven, Bulgaria. Methods: a cross-sectional sero - epidemiological representative population - based survey in the area was made. Enzyme immunoassay method was used for an indirect proof of mumps - specific IgG serum antibodies. 410 people were examined at an average age of 25 (1 to 84). Of these, 250 (61 %) were women and 160 (39 %) - men. Results: Of all test results, the negative were 72 (19 %), the borderline were 12 (3 %), the positive were 182 (44 %), and highly positive were 144 (35 %). The vaccination status showed that 242 (69 %) of all surveyed were immunized with a vaccine against mumps. According to the immunization schedule in Bulgaria, 132 (33 %) people were immunized with monovaccine during the years - 1 intake, 80 (20 %) with trivaccine - 1 intake, and 64 (16 %) - 2 doses.
Conclusion: We believe that despite the specific immunprophylaxis carried out against mumps decades on end, the necessary level of protection leading to its elimination has not yet been reached.
Key words: mumps, seroprevalens, immunprophylaxis

Bogdanka Andric1, Dejan Lausevic2, Marina Ratkovic1, Haki Mavric4
1. Clinic Center of Montenegro, Podgorica, Montenegro
Objectives: Ehrlichiosis are primary recognized as animal's diseases, later as human diseases - Humane Monocytic /HME/ and humane granulocytic /HGE/ ehrlichiosis. The primary vectors in transmission of ehrlichial agents are the different ticks. In Montenegro the first etiological confirmation of Ehrlichiosis in human and animals was in 2008.
Method: In period from 2008 - 2010, we has analyzed 259 the dogs, and 112 patients suspect of the vector borne diseases. The serological confirmation of Ehrlichia canis was in serum of 51 /19,69%/ analyzed dogs, and 19 /17%/ patients, thanks to Indirect Fluorescent Method /IFA/. On examination there were included clinical, laboratory and epidemiological methods.
Results: Non specific symptoms including – febricity, headache, vomiting, mialgia and general weakness but also hepatosplenomegaly, adenopaty, high level of transaminases, leucopenia, anemia, and exanthemas / rare/. In our examined group, respiratory symptoms had 62,5% of patients, 14% appeared with blood disorders. Suspected at Infective Mononucleosis were 38,75%, and 30% of cases had exanthema.
IFA method showed the participation of Ehrlichia canis in 32% of patients. In 68% we detected co-infective participation of other vector borne agents - Rickettsia conorii in 50% cases, Coxiella burneti in 6% of cases, B. burgodrferi in 44% of cases /by Elisa, IIF and FII methods/.  In 35 /68,62%/ analyzed serum of the dogs, we detected co-infective participation of R.conorii and Ehrlichia canis. Only 2 /3,9%/ analyzed serum the dogs, present co/infection of Ehrlichia canis and Coxiella burneti.
Conclusion: Ehrlichiosis is a difficult infectious disease to diagnose because it manifests as an acute undiferential febrile RMSF-like illness eighth few to no physical findings. Co-infections of various tick-borne pathogens transmitted by the same vectors additionally complicated the diagnosis. The preferred drugs for both human ehrlichiosis are doxycicycline. Rifampin may be useful in patients who are unable to take doxycycline /children, pregnant women/. Fluoroquinolones may be useful against ehrlichia, but experience is limited.

Legionella in the Water Distribution Systems?: “Applying Disinfection Methods” (Two Cases’ Report: Paros- and Milos- Primary Health Care Centers)
Valla E. E., G. Koukos*, Fragou A., Gotsi F.
Primary Health Care Center of Milos, “Health Management”. Milos, Greece.  Primary Health Care Center of Paros, “Infection Diseases”, Medical School (University Hospital “ATTIKON”), University of Athens. Greece. Regional Health Department of Paros, Kyklades District (Ministry of Health and Welfare). Paros, Greece.
Introduction: Legionella pneumophila (L.p.) in the water distribution system of a health care unit is a threat to public health -not only for immunosuppressed patients- and an occupational hazard for dentists and all health care workers.
In May 2008, L.p.and spp had been identified in non-acceptable concentrations (>100cfu /ml) in the water distribution systems of two Greek Island’s Primary Health Care Centres (Paros and Milos).
Aim: To present all actions had been taken to disinfect the whole hydraulic networks in order to be safe.
Methods: The dental departments stopped their operation immediately. With the collaboration of the Department of Microbiology, National School of Public Health and the technical advice of expert-engineers:
- the central heating's old boiler was replaced "thermal shock" was applied - the building’s internal network was overheated and flushed for 3 days - network's perchlorination shock (> 0.4 ppm), - all pipes of dental unit and showerheads were replaced - checked \"blind points\" - in Paros: installation of tank chlorination circuit, (liquid clorine ,Cl2) and use of sterile bottled water for the dental unit.
In addition, an automatic water chlorinator (sodium hypochlorite solution, NaOCl) and two UV(Ultra Violet) devices were installed to Milos.
Results: After the procedures were completed the laboratory tests showed that the hydraulic systems were fully disinfected. Four months later, we checked again. Low concentration colonies of L.p. were detected in four of the eleven samples (Paros) and one of sixteen (Milos); but the samples from dental unit remained negative.
Conclusions: All the disinfection methods have been proven to be effective in short term, but long term efficacy has been difficult to be achieved. A successful Legionella prevention program requires routine environmental surveillance cultures and consecutive vigilance. Even though the cost of the appropriate measures to follow up and sustain the quality of the water is very high for small units, such as the PHCC, it is important to strengthen the controls for Legionella, and the measures must also be extended to detect other microorganisms in water (cryptosporidium, etc.).

Mechanism of Action of Newly Synthesized Derivatives of 8-Hydroxy-2-Styrylquinolines in Cell Culture: Evidence for Impact on More than One Human Immunodeficiency Virus - Type 1 Target
A. Hinkov1, V. Atanassov2, S. Dochev3, S. Raleva4, P. Genova-Kalou4, R. Gavazova5, I. Manolov3, A. Pavlov6, S. Chervenkov6, D. Dundarova4, R. Argirova4*
1) Faculty of Biology, Sofia University “St. Kliment Ohridski”, Sofia, 2) Faculty of Chemistry, Sofia University “St. Kliment Ohridski”, Sofia, 3) Faculty of Pharmacy, Medical University, Sofia, 4) National Centre of Infectious and parasitic Disease, Sofia, 5) Institute of Experimental Pathology and Parasitology, Bulgarian Academy of Sciences, Sofia, 6) Trakia University, Stara Zagora, Bulgaria
Objectives: The main complications of Highly Active AntiRetroviral Therapy (HAART) necessitate introduction of new chemical compounds into the HIV/AIDS treatment regimes. Along with inhibitors of reverse transcriptase and protease (RTIs, PIs resp.), integrase inhibitors (INIs) are becoming a standard in clinical practice to maintain the antiviral effect and to prevent emergence of drug resistant mutants. The aim of this work is to present six newly synthesized 2-styryl-8-hydroxyquinolines (designated as initial substances) and five derivatives of them - salts (S), bases(B) and hydrochlorides(H),synthesized after evaluation of anti-HIV activity of the initial substances.
Methods: All the compounds were synthesized and confirmed in Trakia University-Stara Zagora, Bulgaria . MT-4 cells were used for acute infecion with HIV-1 III B and testing of inhibitory effect and.MTT test - to detect survival of infected cells treated /untreated with substances. Supernatants of H9/HTLV III B cell line were used as a source of HIV-1. RT and recombinant RT (rRT) activities were measured by HS-LentiRT Assay (Cavidi, Sweden), protease activity - by home-adapted method based on decrease of product of enzyme reaction between native HIV-1 protease and protease substrate III (Bachem, Germany), IN activity was detected by presence/absence of 2-LTRs in DNA of infected and inhibitor-treated cells.
Results: Two initial 8-SQs protected over 90% of virus-infected cells measured by MTT test. The effect could only partially be explained by RT and rRT inhibition. An deacetylated derivative and a new 8-SQ showed less cytotoxicity , cell protection 100% and 69,9% maximal non-toxic concentration and inhibition of rRT activity not more than 50%. Both 8-SQs showed anti-protease activity (20% and 25% resp.) SQs inhibiting HIV replication were used to pressure the virus to develop mutations in serial passages in MT-4 cells with increasing concentrations:mutants obtained are in process of sequencing.
Conclusion: New anti-HIV 8-SQs with low cytotoxicity were synthesized showing impact on more than one viral targets. However, the anti-HIV RT and protease activity do not fully explain the survival of virus-infected cells in presence of inhibitors. As far as 8-SQs along with other activities inhibit HIV-1 integrase, the mutants are subjected to appropriate studies.

E. Ikonomopoulou, I. Katsarou*, G. Stavropoulou, S. Kapralos, C. Deimente, P. Massoura, A. Regli.
Microbiological unit of General Hospital of Patras, *Renal unit of General Hospital of Zakynthos, Greece.
The Echerichia coli is the most frequent microbial agent in urinary tract infections.
OBJECTIVES: The purpose of this study is to compare the in vitro antibiotic resistance of E.coli in community acquired urinary tract infections between 2001-2002 and 2008 - 2009.
METHODS: A total of 1144 / 912 of E. coli isolates caused community - acquired urinary tract infections during 2001-2002 / 2008-2009 respectively. Isolation was performed by standard method. The identification and the susceptibility test to the antimicrobial agents were performed by the Vitek or the mini API system (Biomerieux).
RESULTS: The resistance of E.coli isolates to antibiotics for 2001-2002 / 2008-2009 respectively was: Ampicillin 32,8% / 35.5%, Amoxicillin / Clavulanic acid 5,0% / 6,0%, Cefalothin 11,6% / 14,0%, Cefuroxime 18,0% / 21,2%, Cefotaxime 1,0% / 3%, Imipenem 0% / 0%, Tobramycin 1,7% / 4,0%, Amikacin 0,9% / 0,8%, Netilmicin 1,0% / 1.6%, Gentamicin 2,1% / 1,8%, Trimethoprim - Sulfamethoxazole 15,8% / 16,2%, Nitrofurantoin 2,4% / 1.6%, Nalidixic acid 8,6% / 12,5%, Ciprofloxacin 7,0% / 10.2%, Norfloxacin 7,0% / 10,2%. The prevalence of ESBL producing urinary E. Coli strains was 1.0% / 3.0% strains respectively.
CONCLUSIONS: The resistance of E. coli strains to ampicillin, cefuroxime, cefalothin nalidixic acid and trimethoprim-sulfamethoxazole increased through the periods. This suggests that the previous antimicrobial agents would not provide adequate initial therapy as empirical choice. The alternative oral therapy may include amoxicillin / clavulanic acid and nitrofurantoin as these antimicrobial agents remain more active. Fluoroquinolones increased their resistance rate through these years probably because of the excessive use.

Molecular epidemiology of HPV infection using a clinical array methodology in Greece
Sotirios Tsiodras, Aris Spathis, Aikaterini Chranioti, John Panayiotides, Asimakis Pappas, Dimitrios Kassanos, Eleftherios Ziogos, Emmanouil Salamalekis, Petros Karakitsos, George Petrikkos.
4th Department of Internal Medicine and Infectious Diseases, University General Hospital Attikon, School of Medicine, National and Kapodistrian University of Athens, Greece
Background: We examined the molecular epidemiology of HPV infection in a large sample of Greek women prior to the implementation of HPV vaccination.
Methods: All women consecutively examined at the outpatient gynecological clinics of several geographical areas in Greece were included. Liquid based cytology was performed and HPV DNA was detected with the use of Papillomavirus Clinical Arrays kits (CLART(R) HPV, Genomica).
Results: Up to January 2009, 2728 women (mean age 42.2 В± 13.3 yrs) were examined and had evaluable cytological results. The prevalence of HPV infection was 49% (95% CI: 47%-50.9%). The prevalence of the most frequently identified HPV genotypes were, HPV 53: 9.3% (95%CU 8.2%-10.5%), HPV 66: 9.1% (95%CI 8.1%-10.4%), and HPV 51: 9.1% (95%CI 8%-10.3%), HPV 16 genotype had a prevalence of 6.8% (95% CI: 5.9%-7.9%), and HPV 18 a prevalence of 3.7%, 95%CI: 3%-4.5%). HPV positivity was associated with younger age (p<0.001), younger age of sexual debut (p<0.001), current and total number of sexual partners (p<0.001), shorter duration of a sexual relationship (p<0.001), while marriage (p<0.001) or multiparity (p<0.001) had a protective effect. HPV DNA positivity for any genotype correlated with both LGSIL (OR 4.7, 95%CI: 3.5-6.3, p<0.001) and HGSIL (OR 17.5, 95% CI: 5.5-56.3, p<0.001). Infection with any of the HPV genotypes 16, 18, 45, 52, 53, 59, 68 and 82 was associated with HGSIL.
Conclusion: High risk HPV types not covered by the current HPV vaccines were most prevalent in this cohort. Infection with non-vaccine HPV types was associated with both low and high grade cytological changes. Follow-up of this cohort will assist in predicting the effect of vaccination with the new HPV vaccines on future screening with HPV based tests for cervical cancer.

In Vitro Activity of Fosfomycin Alone and in Combinations against KPC-Producing Klebsiella pneumoniae Clinical Isolates
M. Souli, I. Galani, G.M Gourgoulis, S.G. Boukovalas, Z. Chryssouli, T. Panagea, H. Giamarellou, K. Kanellakopoulou
4th Department of Internal Medicine, Athens University School of Medicine, Chaidari, Greece
Introduction: The armamentarium of available antibiotics against Enterobacteriaceae producing KPC enzymes is extremely narrow, as extensively drug resistance phenotypes are common for these pathogens. We evaluated the in vitro activity of fosfomycin, a cell wall- synthesis inhibitor, alone and in combinations against KPC- positive K.pneumoniae (Kp).
Methods: Fourteen strains of Kp isolated from different patients were studied. MICs of fosfomycin (FM), meropenem (MER) and gentamicin (GM) were evaluated by agar dilution whereas those of colistin (COL) were evaluated by Etest. The susceptibility to FM was evaluated according to EUCAST breakpoints. KPC production was confirmed by PCR using specific primers. In vitro interactions were determined with time kill methodology with an inoculum of 5X105 CFU/ml. Tubes containing FM (1x, 2x or 4xMIC) in Mueller-Hinton broth supplemented with 25 mg/L glucose-6-phosphate and MER (10mg/L) or GM (10mg/L) or COL (5mg/L) alone and in combination were subcultured at 0, 2, 4, 6 and 24 hours of incubation for viable colony counts. Tubes without antibiotic were included as growth controls. Synergy or indifference was defined as Ві2 or <2log10 CFU/ml decrease of viable counts as compared to the most active single agent.
Results: MIC range of FM, MER, GM and COL was 8->256, 4->256, <2->8 and 0.38-48 mg/L, respectively. FM susceptibility rate was 71.4%. After 24h of incubation bacteria exposed to FM or MER alone showed regrowth. GM was bactericidal against all susceptible isolates and COL was bactericidal against 5 but regrowth was noted for 9 isolates. Synergy with GM was not observed. The combination with MER showed synergy against 5 and indifference against 8 isolates while the combination with COL showed synergy against 3 and indifference against 11 isolates.
Conclusions: There are limited therapeutic options against KPC producing Kp. FM alone although in vitro active against most of the tested isolates, it was not bactericidal. The combinations of FM with MER or COL appeared promising against some of the tested isolates and deserve further clinical evaluation.

Frequency and Antibiotic Susceptibility of Gram(-) Bacteria in ICU Specimens during a 2 Year Period
Vasileios Blioskas, Christina Stefanidou, Eleni Kakasi, Varvara Galanopoulou, Asimoula Koteli*
Microbiology Laboratory, General Hospital \”Agios Pavlos\” ,Thessaloniki, Greece
Objectives: To overview the relative frequency of Gram(-) bacteria isolated in ICU specimens and antibiotic susceptibility over the years 2008-2009.
Material Methods: 250 positive cultures in ICU. The identification of the isolates and the susceptibility testing were performed by the VITEK 2 system (bioMerieuxВ®, France) and susceptibility disc diffusion method according to CLSI guidelines. All ESBL producing isolates flagged by the VITEK 2 were further confirmed by E- test method and double-disk synergy test for screening ESBLs. (SolnaВ®, Sweden). K.pneumoniae ATCC 70603 was used as a positive ESBL-producing control strain.
Results: The specimens were: blood 93, bronchoalveolar lavage 48, wound 42, urine 34, central venous catheter 11, drainage fluid 11, decubitus ulcer 4, bronchial secretion 3, mouth smear 3, pleural fluid 1.
282 microbial species were isolated. 163 (57,8%) were Gram(-).The most frequent Gram(-) species and their percentages among them were: A.baumannii 62 (38%), P.aeruginosa 33 (20,3%), K.pneumoniae 22 (13,5%), E.coli 16 (9,8%), S.marcescens 7 (4,3%), P.mirabilis 6 (3,7%), S.maltophilia 4 (2,5%), E.aerogenes 3 (1.8%), K.oxytoca 2 (1,2%), E.cloacae 2 (1,2%), A.denitrificans 2 (1,2%).
The susceptibility percentages were:
A.baumannii: amikacin 21%, aztreonam 0%, ceftazidime 2%, cefepime 3%, ciprofloxacin 2%, colistin 95%, imipenem 10%, piperacillin/tazobactam 3%, tigecycline 62%, cotrimoxazole 21%
P.aeruginosa: amikacin 67%, aztreonam 58%, ceftazidime 67%, cefepime 68%, ciprofloxacin 61%, colistin 94%, imipenem 59%, piperacillin/tazobactam 84%
K.pneumoniae: amikacin 75%, aztreonam 33%, cefoxitin 35%, ceftazidime 35%, cefepime 35%, ciprofloxacin 45%, colistin 100%, imipenem 55%, piperacillin/tazobactam 50%, tigecycline 56%
E.coli: amikacin 63%, aztreonam 63%, cefoxitin 63%, ceftazidime 63%, cefepime 63%, ciprofloxacin 56%, colistin 100%, imipenem 100%, piperacillin/tazobactam 63%, tigecycline 100%
S.marcescens: amikacin 100%, aztreonam 100%, ceftazidime 50%, cefepime 100%, ciprofloxacin 100%, imipenem 100%, piperacillin/tazobactam 100%, tigecycline 100%
P.mirabilis: amikacin 100%, aztreonam 100%, cefoxitin 100%, ceftazidime 100%, cefepime 100%, ciprofloxacin 80%, imipenem 80%, piperacillin/tazobactam 100%
Conclusion: A.baumannii, P.aeruginosa and K.pneumoniae are the most frequent Gram(-) species isolated in ICU. These have multi-resistance. Good in vitro activity against: A. baumannii have colistin, tigecycline, P. aeruginosa: colistin, piperacillin/tazobactam, K. pneumonia: colistin,amikacin

Antibiotic resistance and virulence factors in opportunistic Gram-negative bacteria
Ivan Mitov, Raina Gergova, Tanya Strateva
Department of Medical Microbiology, Medical Faculty, Medical University of Sofia

Objectives: A total of 372 Gram-negative clinical strains including problematic nosocomial Pseudomonas aeruginosa (n=210), cystic fibrosis (CF) P. aeruginosa (n=47) and Moraxella catarrhalis (n=115) isolated during 2002-2009 were studied to evaluate the current levels of antimicrobial resistance, prevailing resistance mechanisms and distribution of different virulence factors.
Methods: Antimicrobial susceptibilities were tested by E-test. Molecular-genetic investigations were performed using PCR and DNA sequencing.
Resuts: The half of nosocomial P. aeruginosa strains were multidrug-resistant. The most active antipseudomonal agents were: colistin>imipenem>meropenem>ceftazidime>cefepime. High resistance rate (98.26%) to penicillin G, aminopenicillins and first generation cephalosporins was observed among tested M. catarrhalis isolates. Susceptibilities to cefuroxime, cefotaxime and ceftriaxone varied from 86.08% to 97.40%. The prevailing beta-lactamases were BRO-1 (91.3%) and VEB-1 (33.1%) found in M. catarrhalis and P. aeruginosa, respectively. The distribution of virulence genes among all studied P. aeruginosa was as follows: algD (alginate) - 91.1%, pilB (type IV fimbrial biogenesis protein PilB) - 23.8%, nan1 (neuraminidase) - 21.3%, lasB (elastase LasB) - 100%, plcH (haemolytic phospholipase C precursor) - 91.6%, exoS (exoenzyme S) - 62.4%, and exoU (exoenzyme U) - 30.2%. The prevalence of nan1 was significantly higher (P<0.01) in CF isolates (38.1%) than that in non-CF isolates (16.9%). This study revealed a statistically significant difference (P<0.01) between the portion of multidrug-resistant (MDR) nosocomial P. aeruginosa strains containing a large number (more than 5) of virulence genes (38.1%) and the respective part of non-MDR isolates (17.6%). In M. catarrhalis isolates, the frequency of virulence genes encoding outer membrane proteins for adhesion, fat acids transport, complement resistance and vital iron uptake was as follows: ompE - 82.61%, ompCD - 70.43%, and ompB2 - 43.48%. In patients with lower respiratory tract infections the number of isolates with ompB2+ompCD+ompE combination was predominant. In patients suffering from otorrhinolaryngeal infections ompCD+ompE prevailed. In comparison to these groups of patients, the isolates from healthy carriers contained either ompCD or ompE genes only.
Conclusion: Simultaneous determination of virulence factors and antimicrobial resistance is the contemporary approach for examination of the microbiological aspects of infections caused by opportunistic Gram-negative bacteria.

Collateral effects of antiinfectives-from concept to medical activity
Mihai Nechifor
Department of Pharmacology, “Gr. T. Popa” University of Medicine and Pharmacy Iasi Romania
The concept of collateral effects ( CEs)of antiinfectives is a modern and complex notionwich includes not only adverse reactions but also economic, ecological,psychological effects and social influence of this group of drugs.CEs are frecvently omited, not reported or ignorated.There are o lot of problems about the correlation between drug adverse reactions and theirs economic and social impact in clinical practice. It is not clear if Naranjo and Begaud algorithms accurate enough to ascertain the causality of advers effects of antiinfectives. A big question isthe difference between CEs of drugs used in therapy compared to prophylactic use of antiinfectives.In the estimation of the benefit of antibiotic prophylaxis of bacterial infection,the CEs of drugs are frecvently omited.Is not very clear if the drug interactions influence the emergence , incidence, gravity and economic impact ofadverse antiinfective reactions.Not only the presence but also the price paid for advers effects isunderestimated.Our data showed that adverse reactions of antibacterial drugs increased the price of therapy by 20-25 %.We believe That in the evaluation of the ratio benefit/risk of antiinfective therapy should consider all CEs.A new problem is what is the involvement of antiinfectives metabolits in adverse effects of drugs.Some adverse reactions have same incidence and severity in children and in adult patient but not all.Our data showed significant differences between CEs in children versus the adult patients which received antiretroviral drugs.The knowledge of CEs is very important for the progress of antiinfective therapy

Evaluation Of Tigecycline Susceptibility By Disc Diffusion Method in Multidrug-Resistant Acinetobacter spp.
Yilmaz Koz, F. F. *, 1, Tasli, H. 1, Gul-Yurtsever, S. 2, Buyuk, A. 1, Hosgor-Limoncu, M1.
1Ege University Faculty of Pharmacy, Dep. of Pharmaceutical Microbiology, Izmir, Turkey
OBJECTIVES: In recent years, new antibacterial agents are needed for the treatment of infections caused by multidrug-resistant (MDR) Acinetobacter spp. Tigecycline, the 9-tert-butyl-glycylamido derivative of minocycline, exhibits a broad-spectrum of activity against numerous pathogens, including Acinetobacter spp. In this study, we aimed to evaluate microbiological activity of tigecycline for MDR Acinetobacter spp.
METHODS: Between December 2009 to December 2010, 84 MDR Acinetobacter spp. isolates were obtained from Clinical Microbiology Laboratory of Izmir Ataturk Training and Research Hospital. Susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines, by using 15 Ојg tigecycline discs , and zone diameters were evaluated disc diffusion breakpoints for Enterobacteriaceae proposed by FDA (‰19 mm and ‰ 14 mm) and by Jones et al. (≥16 mm and ≤ 12 mm).
RESULTS: It was found that 3 strains (3,57 %) were resistant, 35 strains (41,67 %) were moderately susceptible, and 46 strains (54,76 %) were susceptible to tigecycline when considering breakpoints suggested by FDA, and none of the strains was resistant, 15 strains (17,86 %) were moderately susceptible, and 69 strains (82,14 %) were susceptible according to Jones et al.. Escherichia coli ATCC 25923 was used as control.
CONCLUSION: In our country, antibiotic susceptibility tests are performed generally by disc diffusion method. It is important to decide the evaluation criteria to determine the antibiotic susceptibility properly. To the both two methods, tigecycline showed considerable antimicrobial activity against MDR (including carbapenem-resistant) Acinetobacter spp.. However, the data have to be supported by broth microdilution tests and further studies.

Determination Of Multidrug-Resistant Acinetobacter spp. Biofilm Formation By Two Different Screening Methods
Yilmaz Koz, F. F. 1, Tasli, H. 1, Buyuk, A. *, 1, Gul-Yurtsever, S. 2, Hosgor-Limoncu, M1.
1Ege University Faculty of Pharmacy, Dep. of Pharmaceutical Microbiology, Izmir, Turkey

OBJECTIVES: Multidrug-resistant (MDR) Acinetobacter spp. strains are responsible several types of nosocomial infections including pneumoniae, urinary tract infections. Recent studies showed a positive correlation biofilm formation and antibiotic resistance. Biofilm formation is a significant pathogenic mechanism in device related infections in hospitals. In the present study, we tested biofilm formation in clinic MDR Acinetobacter spp. isolates by two different methods.
METHODS: Between 2009 and 2010, 84 MDR Acinetobacter spp. isolates were collected at the Clinical Microbiology Laboratory of Izmir Ataturk Training and Research Hospital. Biofilm formation was determined qualitatively by tube method and quantitatively using microtitre plate method. Enterococcus faecalis ATCC 29212 was used as positive control.
RESULTS: In our study, qualitative and quantitative test results showed different rate of biofilm production capacity. To the qualitative test, it was determined that 38 strains (45,2 %) were weak/non biofilm producer, 38 strains (45,2 %) were moderate producer, and 8 strains (9.5 %) were strong biofilm producer. However, to the quantitative test, it was described that 34 strains (40,5 %) were weak/non biofilm producer, 13 strains (15,5 %) were moderate producer, and 37 strains (44 %) were strong biofilm producer.
CONCLUSION: Today, treatment of nosocomial infections caused by biofilm producer pathogens is more difficult. As known biofilm formation influences the efficacy of antibiotic and susceptibility of the bacteria. In this study, although qualitative and quantitative test showed different results, it is observed that generally MDR strains are biofilm producer organisms.

Molecular characterization plasmid-mediated quinolone resistance determinants among clinical isolates of Enterobacteriaceae in a Tertiary Hospital in Turkey
Mine Hosgur- Limoncu1, Bayra Erac1, Ayse Nur Yurtman*1, Еuhret Aydemir2
Ege University, Faculty of Pharmacy, Department of Pharmaceutical Microbiology; 2Faculty of Medicine, Department of Microbiology and Clinical Microbiology, Izmir, Turkey
Objectives: The aim of this study was to characterize the prevalance of plasmid-mediated quinolone resistance determinants (qnrA, aac(60)-Ib-cr, qepA) in clinical isolates of Enterobacteriaceae.
Methods: 76 enterobacterial isolates collected in 2004 and 118 enterbacterial isolates in 2008 were assayed for their susceptibility of nalidixic acid and ciprofloxacin with disc diffusion method according to the CLSI ( formerly NCCLS) recommendations. Clonal relation of all nalidixic acid resistant isolates were screened by rep-PCR. aac(60)-Ib-cr, qnrA, qepA genes were analysed by PCR using spesific primers.
Results: Among 69 non-clonally related nalidixic acid-resistant enterobacterial clinical isolates two qnrA, 59 aac(60)-Ib-cr,one qepA positive strains were detected.
Conclusions: This study constitues the first report of the qepA determinant in Turkey which encodes an efflux pump belonging to the major facilitator subfamily (MSF). aac(60)-Ib-cr encodes a variant aminoglycoside acetyltransferase that inactivates the ciprofloxacin. qnrA is resposible for the production of pentapeptide repeat proteins eliciting quinolone resistance by interfering the quinolones activity on bacterial DNA gyrase and topoisomerase IV. There is a very limited number of study on qnrA and aac(60)-Ib-cr determinants in Turkey. Though qnrA shows low prevalence, aac(60)-Ib-cr has a high prevalence in enterobacterial isolates included in this study.
This study was supported as an Ege University Scientific Research Project (09/ECZ/020)

Klebsiella Species: Blood Culture Isolates and Antibiotic Susceptibility in Kosova
* Rr. Bajrami, Gj. Mulliqi, G.Lila, L. Raka, A. Kurti
University of Prishtina, Faculty of Medicine , Pristina , Kosovo ; Department of Microbiology, Institute of Public Health of Kosovo, Pristina, Kosovo
Objectives: The aim of this study was to asses the occurrence of Klebsiella species from bloodstream infections in the University Clinical Center of Kosova and to evaluate their antimicrobial susceptibility patterns.
Methods: A retrospective study was carried out of all blood culture samples presented growth of Klebsiella species from the year 2006 to 2009.The isolates of Klebsiella species were identified using conventional microbiological methods. Antimicrobial susceptibility testing was performed by Kirby Bauer’s disc-diffusion method according to CLSI.The study was conducted at the Department of Microbiology within the National Institute of Public Health of Kosova in Prishtina.
Results: were examined for 849 blood cultures that grew significant microorganisms.Of the total of isolates, 208(24%) were Klebsiella spp.,recovered from blood culture samples obtained from patients with suspected bloodstream infections.The majority of Klebsiella blood culture isolates were from the Center for Premature Children with 95(45,6%),Neonatal Intensive Care Unit 61(29,3),Pediatri 23(11%) and others 29(13,9%).Klebsiella spp.expressed highest susceptibility toward Imipenem(90,3%) and Ciprofloxacin(81,2%). The resistance rate for Trimethoprim-sulfamethoxasole was (87%), Gentamycin (80,2%).
Conlusion: Results from this study show that Klebsiella spp. has developed high resistance towards the tested antimicrobials. These results confirm the necessity of implementation of effective infection control measures as well as proper usage of antimicrobial agents.

Date of close: May 29, 2010

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