Marian Binek

Characterization of Extended-Spectrum-β-Lactamases Produced by Escherichia coli Strains Isolated from Dogs in Poland.

Escherichia coli is a common cause of infections in companion animals. In recent years the increasing prevalence of resistance to β-lactams, including extended-spectrum cephalosporins, antimicrobials frequently used in small animal veterinary practice, was observed in canine isolates of E. coli. The aim of this study was to detect and to characterize extended-spectrum β-lactamases (ESBLs) produced by E. coli isolated from diseased dogs in Poland. Four isolates out of 119 studied (3.4%) were ESBL-positive. They harbored the bla(SHV-12), bla(CTX-M-15), and bla(TEM-116) genes. This study provides the first report of the occurrence of ESBL-producing E. coli in dogs in Poland.

Multidrug resistance in Escherichia coli strains isolated from infections in dogs and cats in Poland (2007-2013).

The antimicrobial susceptibility of Escherichia coli isolates associated with various types of infections in dogs and cats was determined. The studied isolates were most frequently susceptible to fluoroquinolones and the extended-spectrum cephalosporins (ESCs), antimicrobials commonly used in treatment of infections in companion animals. However, an increase in the percentage of strains resistant to β-lactam antibiotics including ESCs was noted between January 2007 and December 2013. The frequency of multidrug-resistant (MDR) E. coli isolation (66.8% of isolates) is alarming. Moreover, the statistically significant increase of the percentage of MDR isolates was observed during the study period. No difference in the prevalence of multidrug resistance was found between bacteria causing intestinal and extraintestinal infections and between canine and feline isolates. Nonhemolytic E. coli isolates were MDR more often than hemolytic ones. Our study showed the companion animals in Poland as an important reservoir of MDR bacteria. These results indicate that continuous monitoring of canine and feline E. coli antimicrobial susceptibility is required. Furthermore, introduction and application of recommendations for appropriate use of antimicrobials in small animal practice should be essential to minimize the emergence of multidrug resistance among E. coli in companion animals.

Characterisation of Staphylococcus aureus isolated from meat processing plants – a preliminary study

Abstract Introduction: Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) belonging to the clonal complex 398 (CC398) emerged recently in livestock as a new type of MRSA, which may cause zoonotic infections. This study presents data on the characterisation of S. aureus isolated from the meat processing plants. Material and Methods: S. aureus was isolated from 90 samples collected in the raw meat warehouse, from devices and surfaces of meat processing plants, and from finished meat products. The isolates were subjected to molecular analysis in order to investigate the presence of enterotoxin genes, the mecA gene, and to verify whether they belong to the clonal complex 398. The genetic relatedness of the isolates was determined using pulsed-field electrophoresis. Likewise, antimicrobial susceptibility was tested. Results: From 21 S. aureus strains isolated, five belonged to the CC398, two of which were recognised as MRSA and three as methicillin-sensitive Staphylococcus aureus (MSSA). The most prevalent enterotoxin genes were seg and sei. Two MRSA CC398 isolates, three MSSA CC398, and one MSSA were classified as multidrug-resistant. Conclusion: The first isolation of MSSA CC398 from beef in Poland indicates contamination of beef by strains belonging to this clonal complex. The occurrence of multidrug-resistant enterotoxigenic S. aureus isolates in the finished meat products constitutes a potential risk for the consumers.

Resistance of canine methicillin-resistant Staphylococcus pseudintermedius strains to pradofloxacin

We investigated in vitro activity of a novel veterinary fluoroquinolone, pradofloxacin, against methicillin-resistant Staphylococcus pseudintermedius (MRSP) isolates and compared with other fluoroquinolones. A total of 38 MRSP isolates were subjected to agar disk diffusion tests for sensitivity to pradofloxacin, orbifloxacin, marbofloxacin, enrofloxacin, and ciprofloxacin. The minimal inhibitory concentration (MIC) values of pradofloxacin, ciprofloxacin, and enrofloxacin were determined. Mutations in the genes encoding DNA gyrase subunit A (GyrA) and topoisomerase IV (GrlA) proteins associated with fluoroquinolone resistance were studied by an analysis of partial sequences of the genes encoding these proteins. Two MRSP isolates were susceptible in disk diffusion and microdilution test to all fluoroquinolones tested, including pradofloxacin. Based on the results of the disk diffusion testing, 33 of 38 isolates showed resistance to pradofloxacin and 3 were intermediate, whereas, by pradofloxacin MIC testing, 35 isolates were classified as resistant and 1 as intermediate. Single alterations in GyrA and GrlA proteins were observed in the 35 resistant isolates and the 1 intermediate isolate (MIC results). These same 36 isolates were also resistant to the other tested fluoroquinolones. The results of the current study showed that MRSP isolates are usually resistant to all fluoroquinolones, including pradofloxacin. Therefore, in routine susceptibility testing to pradofloxacin by disk diffusion, the results should be carefully interpreted for MRSP isolates, especially those resistant to other fluoroquinolones and, in questionable cases, the pradofloxacin MIC should be determined to confirm the susceptibility testing results.

Changes in the population structure of canine methicillin-resistant Staphylococcus pseudintermedius in Poland

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is being reported with an increasing frequency in small animal veterinary practice. The molecular typing of MRSP isolates revealed that the dominating European multidrug-resistant lineage is the sequence type 71 (ST71), associated with staphylococcal chromosomal cassette SCCmec type II-III. However, the recent reports indicated the emergence of other clones. The study aimed to determine the genetic properties of MRSP isolates obtained from dogs in Poland over a ten-year period. A total of 42 clinical MRSP isolates were subjected to multilocus-sequence typing (MLST) and SCCmec typing. MLST typing of 42 MRSP isolates yielded six STs belonging to two major clonal complexes (CCs): CC71 and CC551, associated with SCCmec element II-III and V, respectively. CC71 comprising ST71 and its newly described single locus variant (SLV) ST680. The second dominating CC551was represented by ST551 and newly described SLV ST771. The other, ST258 and ST85 were detected in single MRSP isolates. This is the first report concerning MLST typing of MRSP isolates in Poland. The results confirmed the domination of ST71 among MRSP until 2015, and the emergence of ST551 in 2015. Furthermore, in 2016 ST551 was identified in the majority of the strains, indicating the changes in the population structure of MRSP in Poland. Polish clinical MRSP isolates showed a shift in the population structure during the period of 2007 and 2016. The dominating MRSP lineage until 2015 was multidrug-resistant ST71-SCCmecII-III. The other lineage ST551-SCCmecV emerged in Poland since 2015, and in 2016 was found in the majority of MRSP isolates.