Genotypic characterization ofStaphylococcus aureusisolated from bovines, humans, and food in Indonesia

Abstract

The present study determined the genetic relationships between 41 Staphyloccocus (S.) aureus isolates from bovines, humans, and food using a single enzyme amplified fragment length polymorphism (AFLP) technique. We evaluated the prevalence of staphylococcal enterotoxin (SE) genes and other virulence gene determinants by PCR. The identification of S. aureus was based on culturing and biochemical tests, and by amplifying a specific section of the 23S rRNA gene. PCR amplification of the SE genes (sea, seb, sec, see, seg, seh, and sei) singly or in combination was observed. Most isolates of bovine origin harbored hla (84%) and cap5 (74%), while most isolates from humans harbored hla (73%), cap8 (91%), and fnbA (100%). Strains from food sources were positive for hla (100%), cap5 (100%), and cap8 (64%) unlike isolates from humans or bovines. A single enzyme AFLP analysis revealed a correlation between AFLP clusters of some strains and the source of the isolates The genotypic results of the present study might help to better understand the distribution of prevalent S. aureus clones among humans, bovines, and food and will help control S. aureus infections in Indonesia.

Frequência de genes codificadores de toxinas em Staphylococcus aureus isolados de leite de tanques expansão comunitários

“…Também são reportadas altas frequências em pesquisas internacionais. Na Bélgica, as frequências foram de 98,7% (hla) e 99,1% (hlb) (Ote et al 2011); na Indonésia, as frequências foram de 84% (hla) e 16% (hlb) (Salasia et al 2011) e na China foram encontradas frequências de 85% para a hla e 82% para a hlb (Yang et al 2012). Observou-se que a toxina Hla do S. aureus provoca a morte de um número significativo de eosinófilos in vitro (Prince et al 2012).…”

Genetic Determination and Clonal Relationships of Staphylococcus aureus Isolated from Dairy Cows in Baturraden, Central Java, Indonesia

“…Milk samples were placed in an ice box for transport to the Faculty of Veterinary Medicine, Gadjah Mada University, for isolation of strains. The strains were identified as S. aureus by their properties in mannitol salt agar, Gram staining, and biochemical assays, as described by Ariyanti et al (2011) and Salasia et al (2011). The strains were then identifi ed genotypically by PCR amplifi cation of the 23S rRNA and thermonuclease nuc genes .…”

“…An AFLP analysis was performed according to Boerema et al (2006) and Salasia et al (2011). Genomic DNA (5 μL) was digested overnight (16 h) at 37 o C with 10 U of HindIII (Invitrogen, USA) and 5 mM spermidine trihydrochloride (Sigma, USA) added to a fi nal volume of 20 μL.…”

“…Staphylococcus aureus (S. aureus) is a relatively clonal microorganism that causes subclinical and clinical mastitis (Melles et al 2009;Salasia et al, 2011). This disease causes signifi cant economic losses in dairy farms due to decrease in milk production, shorter lactation times and increased costs for treatment and stock replacement (Halasa et al, 2007;Nielsen, 2009).…”

“…Tato (2012) has proposed that, in order to develop strategies to control S. aureus infection from various countries or areas, we need the ability to map the virulence determinants of bacteria, as well as the capability of the bacteria to survive the heavy pressure of various antibiotics, geographical conditions, and adaptation to environments. Salasia et al (2011) reported that molecular typing of S. aureus and determining the clonal relationship between isolates has proved useful for epidemiological studies. Therefore, mapping the virulence determinant genes and clonal relationship patterns of S. aureus isolates from Baturraden could be important in trying to address the problems there.…”