TABLE OF CONTENT

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1. TULAREMIA. CURRENT STATE AND INVESTIGATIONS IN BULGARIA

V. Tolchkov

ABSTRACT
Tularemia is a very serious zoonotic infectious disease that can be transmitted to humans and cause outbreaks. Control and proper treatment of this disease require early and precise diagnosis. This review summarizes the current state of the problem in Bulgaria and describes worldwide trends in ecology, prevention, diagnosis and therapy. In the last 5 years only few cases of tularemia are reported in our country. However, increased international travel and mobility in the last decades could pose a risk of bringing the disease back.


2. PREVALENCE OF CARBAPENEMASE GENES AMONG 16S RRNA METHYLTRANSFERASE-PRODUCING ENTEROBACTERIACEAE ISOLATED FROM CANCER PATIENTS

I. Ivanov, S. Sabtcheva, E. Dobreva, B. Todorova, Tz. Velinov, V. Borissova, I. Petrova, K. Ivancheva, G. Asseva, P. Padeshki, P. Petrov, T. Kantardjiev

ABSTRACT
The co-existence of carbapenemase and 16S rRNA methyltransferase can cause serious difficulty in treating infections with multidrug-resistant Gram-negative pathogens. In this study we investigated the prevalence of carbapenemase genes among 16S rRNA methyltransferase-producing enterobacteria isolated from cancer patients. A total of 100 non-duplicate clinical isolates of Enterobacteriaceae harbouring methyltransferase genes were studied. Multiplex polymerase chain reaction (PCR) using nine sets of carbapenemase specific primers (VIM, IMP, SIM, GIM, SPM, NDM-1, KPC, GES, OXA-48) followed by sequence analysis of PCR amplicons were used to identify carbapenemase genes. Screening for associated ESBL and AmpC-type genes was carried out by PCR-based assays and PCR products were sequenced. Genotyping, by pulse-field gel electrophoresis (PFGE) of genomic DNA was performed to determine genetic relatedness of carbapenemase-producing isolates. Among the one hundred 16S rRNA methyltransferase-producing enterobacterial isolates, 11 were positive for carbapenemase genes. Of these, 10 ArmA-positive Proteus mirabilis isolates were identified with VIM-1 carbapenemase gene and co-existence of SHV-12 and CMY-99 beta-lactamases. In addition, the NDM-1 carbapenemase gene was identified in one E. coli strain harbouring rtmB methyltransferase gene. The remaining 89 enterobacterial isolates, which were ArmA-positive, did not possess any of the carbapenemase genes studied. This study reports on the low prevalence (11%) of carbapenemase genes among 16S rRNA methyltransferase-producing enterobacteria at the cancer hospital in Sofia, and on the co-existence of VIM-1 carbapenemase gene and ArmA methylase gene in Proteus mirabilis isolates.


3. EXTENDED-SPECTRUM BETA-LACTAMASE SURVEILLANCE IN BULGARIA IN VIEW OF ANTIBIOTIC CONSUMPTION

B. Todorova, T. Velinov, T. Kantardjiev

ABSTRACT
The objective of the present study was to assess the relationship between consumption of ceftriaxone and incidence of extended-spectrum beta-lactamase-producing (ESBL) Escherichia coli and Klebsiella pneumoniae in Bulgarian hospitals between 2007-2013. Cephalosporins and carbapenems on average account for 50% of total hospital antibiotic use, of which ceftriaxone use gradually increases up to 70% during the monitored time period. National surveillance database BulSTAR shows evident increase in ESBL producers from 8% in 2007 to 14% in 2013. Linear regression was used to analyze trends in the annual consumption of ceftriaxone and rates of ESBL confirmed hospital isolates E.coli and K.pneumoniae. We demonstrated statistically significant positive correlation (r=0,87; P=0,01) between these two variables. On average 70 hospitals participated with a total of 25957 isolates yearly. Inferring causal associations between antibiotic use and dynamics in relevant resistance traits is important for identifying possible risk factors for multidrug-resistant infections and outbreaks.


4. LABORATORY-BASED DIAGNOSIS OF GONOCOCCAL AND GENITOURINARY CHLAMYDIAL INFECTIONS

I. Philipova

ABSTRACT
Gonococcal and genitourinary chlamydial infections remain a significant public health concern globally. The incresing burden of these infections, their severe complications that can compromise the general and reproductive health of infected individuals and frequent treatment failures due to increasing antimicrobial resistance require immediate attention. This review focuses on the main aspects and up-to-date recommenations of laboratory-based diagnosis of gonococcal and genitourinary chlamydial infections, on the definitions of a presumptive and definitive diagnosis, on collection and transport of clinical samples, current diagnostic methods (microscopy, culture, nucleic acid amplification tests [NAATs], etc.), antimicrobial resistance testing and molecular genotyping.


5. VIROLOGICAL SURVEILLANCE OF ENTEROVIRAL INFECTIONS AMONG REFUGEES RESIDING WITHIN THE TERRITORY OF BULGARIA, 2013

A. Stoyanova, Z. Mladenova, N. Korsun, P. Teocharov, M. Kojouharova, T. Kantardjiev

ABSTRACT
Owing to the raging conflict in Syria in 2013, the outbreak of polio and refugee wave towards Bulgaria posed a risk for importation of wild poliovirus in our country. Fecal samples were collected from refugee children up to 5 years of age, originating from Syria and other Arab countries and residing on the territory of Bulgaria, as urgent measures required for strengthening the monitoring of polioviruses.

155 fecal samples collected from eight refugee centers were examined at the National Reference Laboratory of Enteroviruses. The assay was performed by utilization of both classical methods and modern molecular techniques (RT-PCR).

From all samples tested, RT-PCR for enteroviruses was positive in 43 (27.74%). Seventeen strains (10.97%) were isolated in cell cultures, including eleven ECHO 3 (7.1 %), one ECHO 29 (0.65 %) and two ECHO 6 viruses (1.29 %). Thirteen specimens showed positive result by both methods.
Three polioviruses were isolated for the first time since the introduction of inactivated polio vaccine in the country: two polioviruses type 3 (1.29 %) and one poliovirus type 1 (0.65 %). Isolated poliovirus strains were sent to the Regional Reference Laboratory in Rome, Italy for intratypic differentiation. PCR and sequence analysis results showed that the isolates are Sabin-like polioviruses.

The largest number of enteroviruses were isolated from children in the age group 2-3 year-olds (41.18%), followed by the groups of 0-1 and 4-5 year-olds with an equal number of isolates (29.41%).

The results indicate the need for continued surveillance of enteroviruses in order to maintain the polio-free status of the country.


6. PARASITIC DISEASES IN BULGARIA IN 2012

I. Rainova, I. Marinova, R. Harizanov, D. Jordanova, I. Kaftandjiev, I. Bikov, N. Tsvetkova

ABSTRACT
Background:
Parasitic diseases are still widespread in Bulgaria and remain as factors adversely affecting human health. The aim of the performed analysis was to determine the status and trends of various parasitic diseases in the country and evaluate the surveillance and control activities conducted in 2012.

Materials: Official surveillance information data was used as well as the the annual reports of parasitological structures in the Regional Health Inspectorates (RHI), which summarize all parasitological situation and activities regarding parasitic diseases in the relevant region.

Results: The results showed that in 2012 a total of 738 155 persons were tested for parasitic diseases and positives accounted for 1.5% (1.63% in 2011). Of the indigenous parasitic diseases helminthozoonoses retain their medical and social importance.The tendency to renewed increase increase in human echinococcosis incidence since 2010, was observed in 2012 as well. In Bulgaria the soil-transmitted parasitoses ascariasis and trichuriasis are annually registered. The prevalence of ascariasis showed no changes in 2012 and there is considerable decrease of trichuriasis prevalence in comparison with the previous years. The distribution of communicable diseases (enterobiasis, giardiasis, hymenolepiasis) remained at the same level as in 2011. Regarding the opportunistic parasitoses, a significant number of tests were carried out mainly for toxoplasmosis and blastocystosis. The registration of autochthonous cases of visceral leishmaniasis is probably due to the presence of a local source of the infection. Among imported from endemic countries parasitic diseases malaria is a leading disease.

Surveillance and control of parasitic diseases are complex and involve case detection and etiological treatment as well as conduction of prophylactic and anti-epidemic measures against the main groups of indigenous and imported parasitic diseases.

Conclusions: Parasitic diseases continue to be an important health problem in Bulgaria. Some of parasitozoonoses are widely distributed in the country; high incidence rates of human echinococcosis are maintained over years and trichinellosis outbreaks are annually registered.
Imported from endemic countries parasitic diseases, and especially malaria, pose a risk of clinical and epidemiological consequences unless strong epidemiological surveillance is in place.

To limit the spread of indigenous parasitic diseases and prevention of the local transmission of imported parasitoses a high level of vigilance and epidemiological surveillance are required.


 

 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
Assoc. Prof. I. Rainova, MD, PhD
Assoc. Prof. P. Teoharov, MD, PhD

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