To determine Salmonella enterica and E. coli, samples (10 g) were prepared by adding 100 mL of phosphate-buffered saline buffer (pH 7.0) and mixed using a stomacher. Then, 0.1 mL of the litter suspension (100) was serially diluted from 101 to 107 dilutions. Serial dilutions were spread-plated on a sterile Petri plate with Difco TM SS agar (Becton, Dickinson and Company, Sparks, MD, USA) and Difco TM Violet Red Bile agar (Becton, Dickinson and Company, Sparks, MD, USA) for counting Salmonella enterica and E. coli, respectively. Plates were incubated at 37°C for 24 h, after which the colonies as average colony forming units (cfu)/g litter were counted at 1 wk through to 5 wk.
Difco ss agar
Manufactured by BD
Sourced in United States
Difco TM SS agar is a selective and differential culture medium used for the isolation and identification of Salmonella species from food, water, and other samples. It inhibits the growth of Gram-positive bacteria and most Gram-negative bacteria, while allowing the growth of Salmonella. The medium contains sodium desoxycholate and brilliant green, which are selective agents, and lactose, which is a differential agent.
Automatically generated - may contain errors
2 protocols using difco ss agar
1
Quantitative Microbial Analysis of Litter
2
Quantifying Gut Bacterial Loads
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To measure LAB, Salmonella enterica, and E. coli loads, fecal samples (200 g) were collected monthly from each pen at five random locations and immediately analyzed. Each fecal sample (10 g) was weighed and placed in a stomacher bag containing 90 mL of sterile saline (0.9%) at a dilution of 1:10. Fecal samples were then plated on Difco MRS agar (Difco, Detroit, MI, USA), DifcoTM SS agar (Becton, Dickinson and Company, Sparks, MD, USA), and DifcoTM Violet Red Bile agar (Becton, Dickinson and Company, Sparks, MD, USA). The MRS agar plates were incubated in a CO2 incubator (Thermo Scientific, Waltham, MA, USA) at 30 °C for 48 h, whereas the SS agar and Violet Red Bile agar plates were incubated for 48 h at 37 °C in an aerobic incubator (Johnsam Corp., Boocheon, Republic of Korea). Visible colonies from the plates were counted, and the number of CFU/g of fecal extract at weeks 0, 30, and 60 was calculated. Microbiological data were transformed to log10.
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