Tryptose sulfite cycloserine agar

The media summarised in Table 2 were used to determine the microbiological contamination of the air.
Moreover, the samples of soil (10 g) and leachate (10 mL) were suspended in 99 mL of sterile saline solution (0.85% NaCl). Dilutions were prepared from 10⁻¹ to 10⁻⁷ in triplicates and then plated onto the above-mentioned media. In addition, TSC (Tryptose Sulfite Cycloserine Agar, Merck Life Science, Warsaw, Poland) was used to quantify the number of sulfate-reducing anaerobes.
All samples were incubated at the following temperature and time: 37 ± 2 °C, and 24–48 h (Enterobacteriaceae, mannitol-positive Staphylococcus sp.), 25 ± 2 °C and 5–7 days (actinomycetes, fungi, xerophilic fungi), 30 ± 2 °C and 48 h (bacteria, sulfate-reducing anaerobes, Pseudomonas fluorescens).
Following incubation, the colonies were counted. The results were expressed in CFU m⁻³ (air) and CFU g⁻¹ (soil) CFU mL⁻¹ (leachate). The arithmetic mean of three independent repetitions was reported as a result.

On day 28 of the experiment, fecal samples (about 50 g) of one male and one female per pen were collected directly from rectum, transferred to sterile falcons, and immediately placed on ice in an insulation box for a maximum of 1 h transportation to the laboratory for further analysis. At the laboratory level, 0.01 g of fresh fecal sample of each animal was taken and placed into Eppendorf tubes supplemented with 990 mL 1× phosphate-buffered saline and 6-fold dilutions were prepared. The samples were plated on agar plates. Bacteria were quantified based on colony-forming units (cfu) counting on the culturing plates (log10 cfu g⁻¹). E. coli were enumerated on eosin methylene blue agar (Merck, Germany) after aerobic incubation at 37°C for 1 day. Salmonella species were enumerated on xylose lysine deoxycholate agar (Merck, Germany) after aerobic incubation at 37°C for 1 day. Clostridium perfringens were enumerated on Tryptose Sulfite Cycloserine agar (Merck, Germany) after anaerobic incubation at 37°C for 1 day. Total anaerobic bacteria were enumerated on nutrient agar (Merck, Germany) after anaerobic incubation at 37°C for 1-2 days. Before statistical analysis, fecal microbiota concentrations were transformed (log).