Molecular identification and further characterization of Arcanobacterium pyogenes isolated from bovine mastitis and from various other origins

Abstract

The present study was designed to identify phenotypically and genotypically 61 Arcanobacterium pyogenes isolated from bovine mastitis and from various other origins. The A. pyogenes isolates showed the typical cultural and biochemical properties of this species and displayed CAMP-like synergistic hemolytic activities with various indicator strains. The species identity could be confirmed genotypically by amplification and sequencing of the superoxide dismutase A encoding gene sodA of reference strains representing 8 species of genus Arcanobacterium and subsequent design of A. pyogenes sodA gene-specific oligonucleotide primer. The A. pyogenes sodA gene-specific oligonucleotide primer allowed, together with previously described A. pyogenes 16S-23S rDNA intergenic spacer region-specific oligonucleotide primer, a reliable molecular identification of all 61 A. pyogenes of various origins. The additionally performed PCR-mediated amplification of 5 known and putative virulence factor encoding genes revealed that 100, 20, 87, 75, and 98% of the A. pyogenes carried the genes plo, cbpA, nanH, nanP, and fimA, which allowed an individual strain characterization. This might help to elucidate the role the putative virulence factors play in bovine mastitis and in various other infections caused by this bacterial pathogen.

Properties and antimicrobial susceptibility of Trueperella pyogenes isolated from bovine mastitis in China

“…Both the nanH and nanP genes also occurred in all isolates recovered from the uterus of dairy cows (Silva and Lobato, 1999). In contrast, other studies reported that the nanH gene was present only in 20.2% (Zastempowska and Lassa, 2012) and 87% (Hijazin et al, 2011) of isolates of bovine mastitis samples. Furthermore, 46 T. pyogenes isolates (92%) carried the fimA gene and most of them (82%) simultaneously harboured the fimC gene, while fimG was found in 38% of the isolates.…”

“…Pyolysin (plo) is a major virulence factor, which is responsible for the lysis of host cells (Jost and Billington, 2005). The presence of the plo gene was demonstrated in all T. pyogenes strains of various origin (Silva and Lobato, 1999;Santos et al, 2010;Hijazin et al, 2011;Zhao et al, 2011). The plo gene was also detected in all of our T. pyogenes strains isolated from bovine mastitis cases.…”

“…The majority (80%) of the isolates displayed a strong positive effect in the synergistic CAMP test performed with S. aureus producing beta-haemolysin. However, variable results (36-100%) have been reported for this test previously (Ülbegi-Mohyla et al, 2009;Hijazin et al, 2011;Rzewuska et al, 2012).…”

“…Trueperella pyogenes is an opportunistic pathogen that causes suppurative diseases including mastitis, abscesses, pneumonia, and lymphadenitis in economically important food animals such as cows and sheep (Ribeiro et al, 2015). The presence of known potential virulence factors of T. pyogenes, such as the haemolytic exotoxin pyolysin (plo), collagen-binding protein (cbp), and factors stimulating adhesion to host cells including fimbriae (fimA, fimC, fimE, fimG) and neuraminidases (nanH, nanP), has been studied in T. pyogenes strains (Silva and Lobato, 1999;Jost et al, 2002;Hijazin et al, 2011;Zhao et al, 2013). Bacterial biofilms, embedded in a self-produced polymer matrix consisting of polysaccharide, protein, and DNA, show an increased tolerance to antibiotics and disinfectants as well as resistance to phagocytosis and other components of the immune system; therefore, they result in chronic infections (Høiby et al, 2010).…”

Isolation and Molecular Identification of Staphylococcus Spp. From Bovine Mastitis Milk Samples.

“…Obtained Sequences were blasted with the GenBank NCBI nr database (http://www.ncbi.nlm.nih.gov/). The highest identity was selected as the identified and according to the results, sequences were submitted for species assignment (Hijazin et al 2011).…”