D glucose

To perform the 11 experiments designed by the CCRD, S. Typhimurium cells recovered from BDL samples—MS cells (Section 2.2 and Section 2.3.1)—at each incubation condition of the CCRD runs were estimated by enumerating surviving cells (Table 1). S. Typhimurium inactivation was calculated as decimal logarithm reductions between the initial inoculum and the recovered cells—Log N₀/N.
The same 11 experimental conditions were used to assess the effect of each exposure condition in the BDL matrix on subsequent SGF S. Typhimurium survival (Table 1). After each experiment, 10 g of the BDL samples were homogenized using the stomacher with 90 mL of SGF and incubated at 37 ± 1 °C for 1 h. The SGF comprised 8.3 g proteose peptone (Sigma-Aldrich, Darmstadt, Germany); 2.05 g sodium chloride; 0.6 g potassium phosphate (Sigma-Aldrich, Darmstadt, Germany) 3.5 g D-glucose (Vetec Química Fina Ltd.a, RJ, Brazil), 0.11 g calcium chloride (Vetec Química Fina Ltd.a), 0.37 g potassium chloride (Tedia Company Inc, RJ, Brazil), 0.1 g lysozyme, and 13.3 mg pepsin (Thermo Fisher Scientific, MA, USA) per liter of distilled water [23 (link),24 (link),25 (link)]. S. Typhimurium enumerations were performed by plating an aliquot of the BDL-SGF cell suspension prior and after the 1 h incubation at 37 °C. S. Typhimurium inactivation was calculated by the logarithm difference from initial and final counts Log N₀/N.

An oral glucose tolerance test was performed at the beginning of supplementation with Ext, S1, and S2, and 3 days before animals were euthanized after 8 h of fasting. Fasting glucose was verified via flow rate (time 0) using a G-Tech^® glucometer (G-Tech Free, Infopia Co., Ltd., Anyang, Gyeonggi-do, South Korea). Thereafter, the animals received D-glucose (Vetec, Duque de Caxias, RJ, Brazil) at 2 g/kg of body weight by oral gavage, and blood glucose was monitored at 15, 30, 60, and 120 min after glucose administration. The area under the curve (AUC) was calculated for each animal and the mean was calculated for each experimental group [51 (link)].

An oral glucose tolerance test (OGTT) was performed three days prior to initiating treatment with the AE and three days prior to euthanasia. The animals were submitted for 8 h of fasting [41 (link)]. The fasting glucose was verified via the flow rate (time 0) using a G-tech glucometer (G-Tech Free, Infopia Co., Ltd., Anyang, Gyeonggi-do city, Korea) with the tail blood. After this, the animals received D-glucose (Vetec, Duque de Caxias, RJ, Brazil) at 2 g/kg of body weight by oral gavage, and the blood glucose was monitored at 15, 30, 60, and 120 min after glucose administration via the tail blood. The area under the curve (AUC) was calculated for each animal, and the mean was calculated for each experimental group [41 (link),50 (link)].