Biofilm Production and Presence of ica and bap Genes in Staphylococcus aureus Strains Isolated from Cows with Mastitis in the Eastern Poland

Abstract

The aim of the study was phenotypic and genotypic analysis of 132 S. aureus strains isolated from mastitis in eastern Poland in respect to their biofilm formation ability. The analysis of the size polymorphism of fragment X in the gene encoding protein A (spa) revealed high genetic differentiation of the analyzed group of isolates. The ability of biofilm formation by the isolates was tested using two phenotypic methods. The Congo Red plate assay was found to be irreproducible and very subjective. More objective results were obtained using the spectrophotometric, microtiter plate assay. Most of the isolates, namely 76/132 (57.6 %) were classified as biofilm producers depending on the value of absorbance in the microtiter plate test. All of the isolates tested were found to possess both icaA and icaD genes, while the bap gene was absent in all strains.

Genotypic and Phenotypic Evaluation of Biofilm Production and Antimicrobial Resistance in Staphylococcus aureus Isolated from Milk, North West Province, South Africa

“…In the current study, biofilm production was assessed by using the MPA, and it was observed that about 90% of S. aureus isolates produced biofilms, out of which the majority (66/70, 94.3%) were classified as "Strong biofilm producers". Similar findings have been previously reported in which most S. aureus strains from milk were biofilm producers [31][32][33][34]. In one of such studies, all 20 S. aureus strains were biofilm producers and 11 out of 20 (55%) were classified as strong biofilm producers [33].…”

“…While Gowishankar and colleagues detected the ica genes in 84.13% of S. aureus isolates in India [35], Avila-Novoa detected the genes in 52.3% of isolates in Brazil [29]. However, the proportion (4, 5.1%) of isolates possessing the icaA and icaD genes associated with slime formation was relatively low in this study, as compared with some previous studies [31,33,34]. These results suggest that strains of S. aureus may present with different capacities to form biofilms based on their source (human or animals) and geographical origin.…”

“…Phenotypic biofilm production in S. aureus has been extensively studied by using the Congo Red plate assay, which is reported to be highly subjective [29][30][31], and as a result, microtitre plate assay (MPA) is considerably used as the gold standard for phenotypic biofilm analyses. In the current study, biofilm production was assessed by using the MPA, and it was observed that about 90% of S. aureus isolates produced biofilms, out of which the majority (66/70, 94.3%) were classified as "Strong biofilm producers".…”

Detection of icaA and icaD genes of Staphylococcus aureus isolated in cases of bovine mastitis in the Republic of Serbia

“…Using the sequencing analysis, Murugan et al [27] have found a high similarity, but some differences in the icaA genes sequences of the 24 S. aureus isolates from conjunctivitis patients. Szweda et al [28] pointed out that some mutations in ica genes are possible and this polymorphism may be the reason for the failed amplifi cation of these genes. Considering that used positive control for icaA gene and 25% investigated isolates were positive during our investigation, we are of the same opinion that the mutations of icaA gene might be the reason for the negative results in our study.…”

Investigation of Biofilm Formation Ability of Coagulase-Negative Staphylococci Isolated From Ready-to-Eat Meat

“…The composition of the reaction mixture used to perform the PCR reaction for all the analyzed genes was as follows: 6.5 µl of PCR Master Mix Plus High GC (A&A Biotechnology, Poland) 0.2 mM of each primer (10 mM), 1 µl of DNA, sterile deionized water to make up the volume of the reaction mixture equals 12,5 µl. The following PCR conditions were used: for icaA and icaD genes -94°C for 4 min, 35 cycles: 94°C for 30 s, 56°C for 30 s and 72°C for 1 min, final elongation at 72°C for 10 min, and for cna gene -94°C for 5 min, 35 cycles: 94°C for 30 s, 56°C for 30 s and 72°C for 40 s, final elongation at 72°C for 7 min (Ote et al 2011;Szweda et al 2012). PCR products were characterized by 1.5% agarose gel (peqGOLD, Peqlab, Germany) electrophoresis in 1× Trisborate-EDTA (TBE) buffer (Bio-Rad, USA), followed by staining with ethidium bromide (Merck, Germany), visualization under UV light and analysis using GeneTools software (Syngene, UK).…”