Macconkey s agar medium

The samples of cecum and ileum content (one bird from each replicate) for microbial analysis were collected by pressing the outer wall of the cut cecum and ileum to push its content into a sterile glass bottle. One gram of cecum and ileum content was blended in 9 mL of sterilized physiological saline solution and homogenized using a mixing bag. Subsequently, these suspensions were serially diluted from 10⁻¹ to 10⁻⁶ using sterilized physiological saline solution. Duplicate plates per bird sample were inoculated with 100 μL of suspension and anaerobically incubated at 37°C.
Bacterial Culture and counts: Lactobacilli were anaerobically assayed using lactobacilli MRS agar (Fisher Scientific, Ottawa, Ontario, Canada) and incubated at 37°C for 48 h. Bifidobacteria counts were performed using Wilkins-Chalgren agar (Oxoid, Nepean, Ontario, Canada) supplemented with glacial acetic acid (1 mL/L) and mupirocin (100 mg/L) extracted from antimicrobial discs (Oxoid, UK). Escherichia coli bacteria were cultured on MacConkey’s agar medium (Oxoid, UK). The numbers of colony-forming units from duplicate plates per bird sample were averaged and the results were expressed as log colony-forming units per gram of intestinal content.

Lactobacillus was anaerobically assayed using lactobacilli MRS agar (Fisher Scientific, Ottawa, ON, Canada) and incubated at 37°C for 48 h. A count of bifidobacteria was performed using Wilkins-Chalgren agar (Oxoid, Nepean, ON, Canada) supplemented with glacial acetic acid (1 mL/L) and mupirocin (100 mg/L) extracted from antimicrobial discs (Oxoid, London, UK) and incubated at 37°C for 20 h. Escherichia coli were cultured using MacConkey’s agar medium (Oxoid, UK). Numbers of colony-forming units from duplicate plates per bird sample were averaged and the results were expressed as log colony-forming units per gram of intestinal content.