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Occurence of virulence genes and biofilm formation in Yersinia enterocolitica isolated from pigs

100%

Kot B., Piechota M., Zdunek E., Borkowska D., Binek T.

Bulletin of the Veterinary Institute in Puławy

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2011

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tom 55

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nr 2

Sixty isolates of Yersinia enterocolitica from pigs belonging to 1A and 4 biotypes, and one non-typeable isolate were investigated in order to determine the occurrence of genes directly connected with pathogenicity of Y. enterocolitica by using a multiplex PCR. The multiplex PCR assay was based on the amplification of the αil, ystA, ystB, and yαdA genes in one reaction. The bacterial adhesion to hydrocarbon test was used to evaluate cells surface hydrophobicity of Y. enterocolitica. Assay of biofilm formation was performed with the use of polystyrene tissue culture plate. In all isolates (n=33) of biotype 1A and in biochemically not typeable isolate, the ystB-specific amplification product of 68 bp was obtained. In all isolates (n=26) of biotype 4, the specific PCR products for ystA gene and αil gene were obtained. In the majority of this biotype isolates (18/26) the specific PCR product for the plasmid yαdA gene was detected. The percentage of isolates with hydrophobic surface of cells was 27.8% and 30.8% within biotype 1A and 4, respectively. All hydrophilic and hydrophobic isolates of Y. enterocolitica adhered to polystyrene, although in different degree. These results suggest that cell-surface hydrophobicity is not important in Y. enterocolitica adhesion and subsequent biofilm formation. The multiplex PCR assay for simultaneous detection the αil, ystA, ystB and yαdA genes, enabled fast evaluation of the potential virulence of isolates and differentiation of the pYV plasmid-bearing Y. enterocolitica isolates, the plasmidless isolates, and biotype 1A isolates.

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Virulence factors and ability of staphylococci from bovine milk and the cowshed environment to biofilm formation

86%

Kot B., Binek T., Piechota M., Wolska K. M., Zdunek E., Platkowska K.

Polish Journal of Veterinary Sciences

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2013

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tom 16

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nr 4

The aim of this study was to examine virulence factors and the ability of S. aureus and CNS species isolated from milk of cows with mastitis to form biofilm, and to compare them with virulence factors of staphylococci from milk of cows without mastitis and cowshed environment. Most of S. aureus strains from cows with mastitis showed haemolytic activity (93.9%), among them 72.7% and 21.2% produced α- and β-haemolysin, respectively. S. aureus from cows with mastitis symptoms produced proteases (above 48%) and esterase (42.4%). The highly significant relationship between the number of S. xylosus strains producing haemolysins (62%) and the origin of these strains from milk of cows with mastitis was observed. The ability to produce proteases was significantly associated with S. sciuri from milk of cows with mastitis. The ability of biofilm formation by staphylococcal strains from milk of cows with mastitis was greater than in strains from milk of cows without mastitis and the difference was significant (p≤0.05). The highest percentage of strains from milk of cows with mastitis were weak biofilm formers (48.6%), while 40% and 11.4% of strains were moderate and strong biofilm producers, respectively. S. xylosus showed the highest ability to form biofilm, while the lowest ability to form biofilm was observed in S. aureus and S. epidermidis. In conclusion, production of exotoxins and enzymes, and ability of biofilm formation shown by many CNS isolated from milk of cows with mastitis symptoms indicates that these features are important in pathogenesis of this disease.

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Antimicrobial resistance and genotypes of staphylococci from bovine milk and the cowshed environment

72%

Kot B., Piechota M., Antos-Bielska M., Zdunek E., Wolska K. M., Binek T., Olszewska J., Gulinski P., Trafny E. A.

Polish Journal of Veterinary Sciences

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2012

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tom 15

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nr 4

Investigation of antimicrobial resistance and genetic relatedness of staphylococci from milk of cows with mastitis and cowshed environment was the aim of this study. Antimicrobial resistance against 14 antimicrobials were determined by using a disc diffusion method. Genetic similarity between the most frequently isolated species was analysed by PFGE (pulsed-field gel electrophoresis). Haemolytic activity, DNase, protease and esterase production was also investigated. Coagulase-negative Staphylococcus species were isolated from 30.8% of milk samples from cows with mastitis. The most frequently isolated species was Staphylococcus xylosus and yield of these organisms was significantly associated with milk of mastitis cows. S. epidermidis was a predominant penicillin-resistant species. High frequency of resistance to lincomycin was observed among isolates of S. sciuri (54.2%) and S. xylosus (25.9%) from cows with mastitis. PFGE (pulsed-field gel electrophoresis) analysis of 29 Staphylococcus aureus isolates showed the presence of 17 PFGE pulsotypes. Isolates of S. sciuri (n=36) had unique PFGE patterns. Some S. xylosus isolates from milk and milker’s hands had the same PFGE pulsotypes, and this observation could indicate that dairyman may be a potential source of the infection. The pulsotype of each of the remaining isolates of S. xylosus suggested that they might have come from common environmental sources; however, these isolates differed in antibiotic resistance pattern or virulence traits. Therefore, knowledge about antibiotic sensitivity pattern and virulence factors of a CNS isolate, besides its genotype, may be informative in tracking the source of the infection.

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Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk

72%

Kot B., Piechota M., Wolska K. M., Frankowska A., Zdunek E., Binek T., Klopotowska E., Antosiewicz M.

Polish Journal of Veterinary Sciences

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2012

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tom 15

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nr 4

The aim of this study was to examine phenotypic and genotypic antimicrobial resistance of staphylococci from milk samples from cows with subclinical and clinical mastitis and from cows without mastitis symptoms to methicillin, tetracyclines, macrolides and lincosamides (ML). Of 207 strains, including 34 S. aureus and 173 coagulase-negative staphylococci (CNS), 11 (6.4%) CNS strains were phenotypically resistant to methicillin. The mecA gene was detected by PCR only in two S. xylosus strains and one strain of S. epidermidis and S. simulans. No methicillin-resistant S. aureus strains were observed. In methicillin-resistant strains with mecA, gene resistance to other investigated antibiotics was not observed. Phenotypic resistance to tetracycline was detected in 11.0% of CNS strains and 47.4% of them carried the tetK gene. Of 173 CNS strains studied, 27 (15.6%) were resistant to at least one ML antibiotic. The resistance gene ermC was detected in 55.5% of the 27 ML-resistant strains. The ermA and ermB genes were detected in 14.8% and 11.1% of ML-resistant CNS strains, respectively. Antimicrobial resistance to methicillin, tetracyclines and macrolides was detected more frequently in staphylococcal strains from clinical mastitis compared to animals with subclinical symptoms and without mastitis, while the resistance to lincosamides showed a similar frequency in all groups of cows. In conclusion, CNS species from bovine milk differ in phenotypic and genotypic antimicrobial resistance profiles, and the use of PCR technique alone for the detection of methicillin, macrolide, lincosamide and tetyracycline resistance in CNS from cattle is not reliable.

Ograniczanie wyników

3 Polish Journal of Veterinary Sciences

1 Bulletin of the Veterinary Institute in Puławy

4 Binek T.

4 Kot B.

4 Piechota M.

4 Zdunek E.

3 Wolska K. M.

1 Antos-Bielska M.

1 Antosiewicz M.

1 Borkowska D.

1 Frankowska A.

1 Gulinski P.

1 Klopotowska E.

1 Olszewska J.

1 Platkowska K.

1 Trafny E. A.

1 2013

2 2012

1 2011

Occurence of virulence genes and biofilm formation in Yersinia enterocolitica isolated from pigs

Virulence factors and ability of staphylococci from bovine milk and the cowshed environment to biofilm formation

Antimicrobial resistance and genotypes of staphylococci from bovine milk and the cowshed environment

Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk