Mrs agar with tween 80

The microbiological parameters (i.e., total bacteria count (TBC), lactic acid bacteria count (LABC), total enterobacteria count (TEBC), and mould/yeast count (M/Y)) of the prepared marinades and the treated LM were evaluated. For this evaluation, 10 mL and 10 g of a sample were homogenised with 90 mL of saline (0.9%). Then, serial dilutions of 10¹–10⁷ with saline were used for the sample preparation.
The LAB counts were determined on MRS agar with Tween-80 (Biolife, Milano, Italy) using standard plate count techniques. The procedures were described in detail in ISO 15214:1998 [65 ].
The total bacteria counts (TBCs) were determined on plate count agar (Biolife, Milan, Italy), as described in ISO 4833-2 [66 ].
The total enterobacteria counts (TEBCs) were determined on violet red bile glucose agar (Oxoid Ltd., Basingstoke, UK), as described in ISO 21528-2 [67 ].
The yeast and mould counts were determined on Dichloran rose Bengal chloramphenicol agar (Liofilchem, Milan, Italy), as described in ISO 21527-2 [68 ].
The total bacteria counts were calculated and expressed as the log₁₀ of colony-forming units (CFU·mL⁻¹ and g) of the samples.

The microbiological parameters of the samples, including TBC, LAB, TEC, and M/Y counts, were evaluated. A 10 g and 10 mL sample was homogenized in 90 mL of a 0.9% sodium chloride solution for this evaluation. The sample was then prepared using saline serial dilutions ranging from 10¹ to 10⁷. The M/Y viable counts were measured on Dichloran rose Bengal chloramphenicol agar (Liofilchem, Milan, Italy); TEC was measured on violet-red bile glucose agar (Oxoid Ltd., Basingstoke, UK); TBC was measured on plate count agar (Biolife, Milan, Italy); and LAB viable counts were measured on MRS agar with Tween-80 (Biolife, Milano, Italy). Section 2.2 provides standards for assessing microbiological parameters.

The contents of the jejunum and colon were used for determination of the count of Enterobacteriaceae, Escherichia coli, and Lactobacillus spp. The initial and serial dilutions were made in peptone saline diluent (Maximum Recovery Diluent, Biolife) according to ISO 6887-1:1999. For the isolation and enumeration of Enterobacteriaceae and Escherichia coli, Violet Red Bile Glucose agar (Biolife) and Tryptone Bile X-Gluc agar (Biolife) were used according to ISO 21528-2:2007 and ISO 16649-2:2007, respectively. For isolation and enumeration of Lactobacillus spp., MRS agar with Tween 80 (Biolife) was used according to the manufacturer's instructions for this medium. The colony-forming units were calculated and expressed as log₁₀ colony-forming units per gram of digestive contents.