By referring to the protocols of AOAC (25 ), the dry matter (DM; method 930.15), ash (method 942.05), crude protein (CP; method 2001.11), ether extract (EE, method 920.39), neutral detergent fiber (NDF; method 2002.04), and acid detergent fiber (ADF; method 973.18) in the feed and feces were analyzed. The contents of calcium (Ca) and phosphorus (P) in the basal ration were assessed as described previously (23 (link), 26 (link)). The measurement for pH and the concentrations of ammonia nitrogen (NH3-N) and volatile fatty acid (VFA) of the rumen fluid was performed using methods depicted in precedent studies (19 (link), 27 (link)). The acid-insoluble ash was used as the internal marker to estimate the apparent digestibility of nutrients (28 ).
The amount of lipopolysaccharide (LPS) endotoxin in the rumen fluid and plasma, and the levels of the inflammatory mediators in the serum were evaluated as previously described (26 (link)). As to the assessments of biochemical blood parameters, a Roche Cobas automatic biochemistry analyzer (c311, Roche Ltd., Basel, Switzerland) and corresponding specific kits (Roche Ltd., Basel, Switzerland) were used, according to the manufacturer’s instructions and prior report (29 (link)).
The amount of lipopolysaccharide (LPS) endotoxin in the rumen fluid and plasma, and the levels of the inflammatory mediators in the serum were evaluated as previously described (26 (link)). As to the assessments of biochemical blood parameters, a Roche Cobas automatic biochemistry analyzer (c311, Roche Ltd., Basel, Switzerland) and corresponding specific kits (Roche Ltd., Basel, Switzerland) were used, according to the manufacturer’s instructions and prior report (29 (link)).