Detection of point mutations in the gyrA gene of Helicobacter pylori isolates in Slovenia.

BACKGROUND/AIMS: The prevalence of antibiotic-resistant H. pylori clinical isolates is a growing concern. So far, fluoroquinolones have not been used to treat H. pylori on a large scale, but recent studies have reported a high rate of quinolones-resistance in H. pylori too. The aim of our study was to asses the mechanism of resistance to ciprofloxacin in H. pylori clinical isolates from patients living in Slovenia. METHODOLOGY: Out of 397 H. pylori clinical isolates, obtained in the period 1997 to 2004, 33 (8.3%) ciprofloxacin-resistant H. pylori isolates were recognized. DNA sequences of the gyrA gene were determined and translated into amino acid sequences. RESULTS: Based on the results of this analysis, various point mutations in the ciprofloxacin-resistant clinical isolates were revealed. The most common mutations in H. pylori gyrA gene were found at codons corresponding to Asp91 (57.6%) and Asn87 (36.4%). Sequence analysis revealed amino acid substitutions also at codons Ala97 to Val, Ala129 to Thr and a double substitution at Asn87 to Lys and Val107 to Ile. CONCLUSIONS: These results suggest that H. pylori resistance to ciprofloxacin is already present in the Slovenian population and that it seems to be mediated through amino acid substitutions in the gyrA gene. Furthermore, the results obtained from the study also demonstrate no significant association between the type of gyrA mutation and the ciprofloxacin MIC level.

Canine babesiosis in Slovenia: molecular evidence of Babesia canis canis and Babesia canis vogeli.

Canine babesiosis, caused by intraerythrocytic Babesia spp., is a tick-borne disease of worldwide importance. No information on canine babesiosis has been documented in Slovenia. Therefore, 238 dogs admitted to the Small animal clinic in Ljubljana from the years 2000 to 2002 were tested for the presence of babesial parasites in the blood. Based on clinical, microscopic and molecular investigations, 14 dogs (5.9%) were determined as being infected with babesiae. Clinical signs relating to acute haemolysis, fever, anorexia, depression and haematological abnormalities such as anaemia and thrombocytopenia were noticed in most of the 14 infected dogs. The morphology of the parasites was indicative of Babesia canis infection. Two subspecies were detected, namely B. canis canis (11 dogs, 4.6%) and B. canis vogeli (3 dogs, 1.3%) using PCR and subsequent sequence analysis of portions of nns rRNA gene. In addition, based on nucleotide sequence analysis, the 11 isolates of B. c. canis could be subdivided into three groups, whereas the three B. c. vogeli isolates were genetically identical. The results of this study demonstrate the presence of canine babesiosis due to B. c. canis and B. c. vogeli in Slovenia.