An approximately 10 g meat sample was weighed in a stomacher bag (VWR International, Darmstadt Germany), diluted 1:10 gravimetrically in peptone water (Carl Roth, Karlsruhe, Germany) and homogenized for 90 s (Stomacher, IUL instruments, Barcelona, Spain) as described in a recent study [21] (link). Samples were serially diluted in peptone water and plated in duplicate on Plate Count agar (AppliChem, Darmstadt, Germany) for the detection of the total viable counts and on Baird Parker agar (Carl Roth, Karlsruhe, Germany), and then mixed with 10 mL of 1% potassium tellurite solution (Sigma Aldrich, Steinheim, Germany). Therefore, 100 µL aliquots of 1:10 dilutions were spread-plated or 50 µL aliquots of 10 -2 -10 -4 dilutions were spiral-plated using an automated spiral plater (Don Whitley Scientific Limited, West Yorkshire, UK). Plate Count agar was incubated for 48 h at 37 °C and Baird Parker agar was incubated for 72 h at 37 °C. Colony forming units (cfu) were measured using a colony counting device (aCOLyte, Synbiosis, Cambridge, UK). Additional sampling was carried out in weeks 1, 2 and 9 to track microbial changes during processing and storage.
Baird parker agar
Manufactured by Carl Roth
Sourced in Germany
Baird Parker agar is a selective and differential culture medium used for the isolation and identification of Staphylococcus aureus from clinical and food samples. The medium contains egg yolk and potassium tellurite, which inhibit the growth of most other bacteria, allowing for the selective growth of Staphylococcus aureus.
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2 protocols using baird parker agar
1
Meat Microbiological Analysis Protocol
2
Staphylococcus Carriage in Kenyan Shelter Dogs
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Diagnostic specimens from all dogs (N = 167) housed at the animal shelter were collected on the 27th June 2015 in Nairobi, Kenya by veterinarians using the TRANSWAB® Amies swabs. Nasal specimens were collected by inserting the swabs up to 10 cm into the nasal cavity of the animals, which were restrained without sedation for the procedure. Wound infections were also swabbed when present. The Staphylococcaceae strains were isolated at the laboratories of the International Livestock Research Institute (ILRI) using standard methods without selective enrichment (44 ). Briefly, each swab was streaked on Baird Parker Agar (Carl Roth) plates and incubated at 37°C overnight. One to two suspicious colonies per plate were picked, expanded in LB media (Carl Roth), and stored as glycerol stocks at −80°C before being shipped for further characterization to the Institute of Veterinary Bacteriology (IVB) in Switzerland. Strains were streaked onto Trypticase Soy Agar (TSA-B; Becton Dickinson) with 5% sheep blood (TSA-B; Becton Dickinson) and incubated at 37°C overnight. Species designation was assigned using MALDI-TOF MS (Bruker) as previously reported (30 (link)) followed by metabolic phenotyping (see below). Data on strains investigated in this study are provided in Dataset S1.
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