TABLE OF CONTENT

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1. DISSEMINATION OF CTX-M EXTENDED SPECTRUM BETA-LACTAMASE PRODUCING ISOLATES OF KLEBSIELLA PNEUMONIA IN FOUR HOSPITALS IN SOFIA AND PLEVEN

R. Markovska, E. Keuleyan, D. Ivanova, B. Markova, J. Proevska, M. Leseva, I. Schneider, A. Bauernfeind, I. Mitov

ABSTRACT
Objectives:
To prove ESBL production in acollection of isolates Klebsiella spp with reduced susceptibility or resistant towards cephalosporins third generation, which were recovered during the period 2007-2009 in four hospitals in Sofia and Pleven. To characterize ESBL types, antibiotic susceptibility of the producers as well as to detect possibility for conjugative plasmid transfer and clonal relatedness of the strains.
Materials and methods: A total of 44 clinically significant isolates Klebsiella pneumoniae and 1 Raoultella terrigena with reduced susceptibility or resistant to IIIrd generation cephalosporins were studied. Antimicrobial susceptibility, ESBL-type and transferability of resistance determinants were analyzed. The epidemiology typing was performed by ERIC -1, 2A PCR.
Results: Production of ESBL was confirmed in 42 studied strains. The CTX-M group ESBL was the predominant one (98%). Only one strain, isolated in 2008, was SHV-12 producer. CTX-M-3 enzymes were found in 45% of the isolates. Among them we have detected two clones with one of them appearing in all centers in Sofia and the other in a hospital in Pleven, each representing 50% of isolates. CTX-M-15 positive isolates (53%) belonged to seven clones with 1 to 7 members. The high rate of conjugation transfer in the collection of CTX-M-3 producers (89%) was detected. In the case of CTX-M-15 producers the rate was 45%. The ESBLs producers demonstrated high resistant rates to tobramycin, gentamicin and amoxicillin/clavulanic acid.
Conclusions: On the base of these results it can be concluded that clonal dissemination most likely plays the important role for the disseminations of CTX-M-3 and 15 producers, while plasmid transfer is associated with distribution of CTX- M-3 enzymes among Klebsiella strains.


2. FIVE YEARS OF ANTIBIOTIC CONSUMPTION IN BULGARIA. SYNOPSIS OF MAJOR TRENDS IN 2007-2011

Tz. Velinov, B. Todorova, T. Kantardjiev

ABSTRACT
Resistance to antibiotics is a major public-health problem and antibiotic use is being increasingly recognised as the main selective pressure driving this resistance. Our aim was to assess outpatient, hospital and total use of antibiotics in Bulgaria during the period 2007-2011.
Materials and Methods: Consumption data were obtained from the pharmaceutical marketing provider IMS Health as number of packages for each antimicrobial product for systemic use. We calculated antibiotic consumption in the three sectors as the number of defined daily doses (DDD) per 1000 inhabitants per day, according to WHO anatomic therapeutic chemical classification and DDD measurement methodology.
Results: Overview of total consumption shows slight decrease between 2008-2010 from 24.6 to 22.5 DDDs/1000 inhabitants per day, respectively. The most frequently used antimicrobials in primary care are arranged in a similar way as in the results for the total consumption. The penicillin group ranks first, followed by MLS (macrolides and lincosamides) and cephalosporin groups. Hospital consumption results have different pattern of the most widely used groups. Almost half of the total hospital consumption is in favor of the cephalosporins with measured values distributed between 0.8 and 1.1 DDDs/1000 inhabitants per day. Second are penicillins and after them – fluoroquinolones.
Conclusion: We measured beta-lactam antibiotics as the largest proportion of antibiotic consumption in the three sectors. An unfavourable trend during the five years of our monitoring is the increasing percentage of broad-spectrum cephalosporins used in Bulgarian hospitals.


3. HUMAN LEPTOSPIROSIS IN BULGARIA – CLICNICAL, EPIDEMIOLOGICAL AND SEROLOGICAL ASPECTS OF INFECTION, 2010-2011

E. Taseva, I. Christova, T. Gladnishka, I. Trifonova, V. Ivanova

ABSTRACT
In the last years, a decrease in the number of registered cases of leptospirosis in our country was observed. This study was focused on the analysis of the circulating serovars of the genus Leptospira and their distribution, and the epidemiological characteristics of the laboratory confirmed cases of human leptospirosis, 2010-2011. A total of 28 cases of leptospirosis were registered and laboratory confirmed by a microagglutination test in Bulgaria in this period. The highest incidence was observed in the following areas : Shumen (25%), followed by Sofia-city (14.29%) and Pleven (10.71%). Infected people were mainly elderly men. The age groups 40-49 years - 7/28 and 60-69 years - 7/28 were mainly affected, respectively (25%). The infection is acquired mainly through recreational activities, mainly fishing - (47.37%). The socio-economic structure and the seasonal distribution of the disease were also analyzed. The clinical spectrum of the symptoms was analyzed in 19 patients with leptospirosis (67.86%). Fever and jaundice were the most frequently reported symptoms in 18/19 patients (94.74%) and in 14 patients (73.37%) resp. The analysis of the serological data showed that the serovars causing leptospirosis belong to five serogroups. The leading serogroup was Leptospira icterohaemorrhagiae 14/28 (50%), followed by serogroups Sejroe (21.43%) and Australis (14.29%). Analysis of the results of this two-year period showed that leptospirosis in Bulgaria is an infection with limited distribution, but the risk of leptospirosis should not be underestimated.


4. EVALUATION OF A NEW MOLECULAR TEST FOR THE IDENTIFICATION OF DRUG RESISTANCE IN MYCOBACTERIUM TUBERCULOSIS CLINICAL ISOLATES

I. Bergval, S. Sengstake, E. Bachyiska, N. Brankova, K. Tankova, V. Levterova, A. Ivanova, Y. Atanasova, S. Yordanova, E. Abadia, C. Sola, M. Akhalia, R. Aspindzelashvili, D. van Soolingen, R. Anthony, T. Kantardjiev, S. Panaiotov

ABSTRACT
Mycobacterium tuberculosis is a clonal organism, once a single nucleotide polymorphism (SNP) is introduced into the genome it will be passed on to the next generation. Drug resistance in M. tuberculosis strains is to a great extent caused by the presence of SNPs in genes conferring resistance to tuberculostatics. SNPs can be used as discriminative markers for drug resistance as well as to generate robust phylogenetic trees. SNP based species identification is also possible. A new molecular method was recently developed for the analysis of drug resistance and strain typing of M. tuberculosis strains, based on the detection of informative genetic markers such as deletions or single nucleotide polymorphisms (SNPs).
The method evaluated in this study, multiplex ligation-dependent probe amplification (MLPA) [Schouten, 2002] contains three essential steps, a hybridization step followed by a ligation step and an amplification step. The final read-out is performed on the MAGPIX device using the Luminex xTAG bead assay. The multiplexing capacity of the assay allows the simultaneous analysis of up to 47 different molecular markers within one sample and therefore provides complex information from a single assay. 13 markers provided information about resistance to first- and second line drugs based on the detection of specific mutations in the bacterial genome. We have analysed DNA isolated from bacterial culture by MLPA and compared the results to the current molecular method for the detection of drug resistance in TB strains, the MTBDRplus and MTBDRsl assays (HAIN Lifesciences).


5. VACCINATION AGAINST HPV IN THE BULGARIAN ARMY MILITARY FEMALE PERSONNEL

A. Galev, A. Kanev, E. Penkov, A. Nacheva

ABSTRACT
On the initiative of representatives of the Ministry of Defence and under the guidance of experts from the Military Medical Academy in May 2009 a program to prevent the spread of human papillomavirus (HPV) among military female personnel has been started. The program covers voluntary female military personnel and includes clinical examination by a gynecologist with sampling for direct microscopy and some of them for Real-Time Polymerase Chain Reaction for HPV serotypes 16 and 18.
Women with negative clinical and laboratory findings were vaccinated immediately. Those with abnormal examinations underwent treatment and after re-examination their participation in the program was refined in each case. The vaccines provided were Cervarix, production of GlaxoSmithKline and Silgard, production of Merck Sharp&Dohme. The program includes five stages: 1) Preparation, 2) Information lectures, 3) Clinical examination, 4) Vaccination and 5) Clinical and laboratory monitoring of vaccinated individuals. For a period of two and a half years - from 2009 until the May 2012 SCMEH - MMA and Preventive Medicine Teams in Pleven, Varna, Sliven and Plovdiv have vaccinated with full course of three doses 480 women (of 631 examinated ) - 396 with Cervarix and 84 with Silgard. The age of the women was 18 to 30 (on the average 27.4 years) with 472 (of 607 tested) and 30 to 40 (on the average 36, 8 years) in 8 of them (of 24 tested). The immunization course was interrupted in 25-female militaries due to pregnancy in 21 women, 1 because of breast cystosis, 1 because a discharge from the armed forces and 2 had not appeared for personal reasons.


6. CLINICAL, EPIDEMIOLOGICAL AND LABORATORY CHARACTERISTICS OF HEPATITS A OUTBREAK IN THE VILLAGE OF ISKRA (PLOVDIV REGION), JANUARY-MARCH 2012

N. Vatev, A. Petrov, M. Troyancheva, M. Stoycheva, V. Georgieva

ABSTRACT
Goal:
To analyze the clinical, epidemiological and laboratory characteristics of the patients from Iskra village with VHA treated in the Clinic for Infectious Diseases during the period January-March 2012.
Materials/methods: For the period considered a total of 225 VHA patients were hospitalized. Of these, 125 were associated with the epidemic outbreak in the village of Iskra. The diagnosis was verified by anti HAV IgM. Routine laboratory methods, clinical observations and epidemiological analysis were applied.
Results: Males were 70 (62.9%) and women - 55 (37.1%). Thirty-three of the study subjests are not permanent residents in the locality, but have an epidemiological link. The index of affected contacts in the existing outbreaks of infection and the risk of occurrence of the disease during outbreaks have been assessed. The average hospital stay was 10 days. The disease was manifested in mild and moderate form. Cholestasis component was registered in 3 of the cases. So far no relapses have been registered.
Conclusion: The studied epidemic outbreak is another proof of the endemic-epidemic spread of VHA in Bulgaria. Therefore preventive measures and disease control need to be improved.


7. NOSOCOMIAL INFECTIONS, CAUSED BY CANDIDA SPP. – FREQUENCY AND SPREAD DYNAMICS DURING THE PERIOD 2000-2011

N. Ribarova, Y. Mitova, S. Angelova, A. Gladilova

ABSTRACT
During the period 2000 - 2011 genus Candida organisms had a significant share of the etiological structure of nosocomial infections in Bulgaria - an average of 4.14%. Spread dynamics of these pathogens tended to increase from 2.95% in 2000 to 5.13% in 2011. Over 40% of the infections caused by Candida spp. were due to C.albicans, with an increase of its relative share to 52.17% in 2011.
Anesthesiology and Intensive Care Units took leading positions in the spread frequency of Candida nosocomial infection with 51.09% of all registered Candida Nosocomial Infections (NI). Microorganisms of genus Candida were the most common agents of urinary (34.43%) and respiratory (33.29%) system infections.


 

Editor-in-Chief
Prof. T. Kantardjiev, MD, DSc
Editorial Board
Acad. B. Petrunov, MD, DSc
Prof. I. Christova, MD, DSc
Prof. M. Kojuharova, MD, PhD
Prof. R. Kurdova, MD, PhD
Assoc. Prof. I. Rainova, MD, PhD
Assoc. Prof. P. Teoharov, MD, PhD

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