Extrusion of lupines with or without addition of reducing sugars: Effects on the formation of Maillard reaction compounds, partition of nitrogen and Nε-carboxymethyl-lysine, and performance of dairy cows.

The extrusion of leguminous seeds induces the formation of Maillard reaction compounds (MRC) as a product of protein advanced glycation and oxidation, which lowers protein degradability in the rumen. However, the quantitative relationship between the parameters of pretreatment (i.e., addition of reducing sugars) and extrusion, and the formation of MRC has not been established yet. Moreover, the fate of the main stable MRC, Nε-carboxymethyl-lysine (CML), in the excretory routes has never been investigated in ruminants. We aimed to test the effects of the temperature of extrusion of white lupines with or without addition of reducing sugars on the formation of MRC, crude protein (CP) degradability in the rumen, N use efficiency for milk production (milk N/N intake), and performance of dairy cows. Two experiments with a replicated 4 × 4 Latin square design were conducted simultaneously with 16 (3 rumen-cannulated) multiparous Holstein cows to measure indicators of ruminal CP degradability (ruminal NH3 concentration, branched-chain volatile fatty acids), metabolizable protein supply (plasma essential AA concentration), N use efficiency (N isotopic discrimination), and dairy performance. In parallel, apparent total-tract digestibility of dry matter, organic matter, neutral detergent fibers, N, total Lys and CML, and partition of N and CML were measured with 4 cows in both experiments. The diets consisted on a DM basis of 20% raw or extruded lupines and 80% basal mixed ration of corn silage, silage and hay from permanent grasslands, pelleted concentrate, and a vitaminized mineral mix. Expected output temperatures of lupine extrusion were 115°C, 135°C, and 150°C, without and with the addition of reducing sugars before extrusion. The extrusion numerically reduced the in vitro ruminal CP degradability of the lupines, and consequently increased the predicted supply of CP to the small intestine. Nitrogen balance and urinary N excretion did not differ among dietary treatments in either experiment. Milk yield and N use efficiency for milk production increased with extrusion of lupines at 150°C without addition of reducing sugars compared with raw lupines. Nitrogen isotopic discrimination between dietary and animal proteins (the difference between δ15N in plasma and δ15N in the diet) were lower with lupines extruded at 150°C without and with addition of reducing sugars. Regardless of sugar addition, milk true protein yield was not affected, but milk urea concentration and fat:protein ratio were lower with lupines extruded at 150°C than with raw lupines. In the CML partition study, we observed that on average 26% of the apparently digested CML was excreted in urine, and a much lower proportion (0.63% on average) of the apparently digested CML was secreted in milk, with no differences among dietary treatments. In conclusion, we showed that the extrusion of white lupines without or with addition of reducing sugars numerically reduced enzymatic CP degradability, with limited effects on N partition, but increased milk yield and N use efficiency at the highest temperature of extrusion without addition of reducing sugars.

Effects of pretreatment with reducing sugars or an enzymatic cocktail before extrusion of fava bean on nitrogen metabolism and performance of dairy cows.

The aim of this study was to determine the effects of pretreatment with reducing sugars or with an enzymatic cocktail before extrusion of fava bean on intake, milk yield and composition, N partitioning, and plasma and ruminal parameters. The main hypothesis was that these pretreatment conditions would allow better N protection in the rumen compared with classic pretreatment before extrusion, thanks to an increase of sugar proportion, either exogenous or endogenous with enzymatic actions. Sixteen Holstein cows were used in a 4 × 4 Latin square design experiment. Cow were fed a diet with a 56:44 forage to concentrate ratio and containing 16.2% of crude protein (CP; dry matter basis). Concentrate consisted of fava bean:linseed blends (90:10%, raw basis) distributed either raw, extruded after pretreatment without additive, extruded after pretreatment with reducing sugars, or extruded after pretreatment with an enzymatic cocktail. The experimental blends provided 53% of total CP in the diet. Intake, milk yield and composition, ruminal pH, volatile fatty acids and ammonia kinetics, apparent total-tract nutrient digestibilities and N partitioning, Maillard compounds in feed and feces, plasma AA, and 15N natural enrichment were measured. Data were analyzed using analysis of variance according to the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Extrusion without additive during pretreatment led to higher contents of Maillard compounds in the blend, a lower enzymatic CP degradability, a numerically lower ammonia content in ruminal fluid, and a trend of higher plasma EAA concentration, all suggesting a decrease in degradability of proteins in the rumen, and a subsequent increase in metabolizable protein supply. Compared with pretreatment without additive, adding reducing sugars or an enzymatic cocktail during pretreatment led to an increase in Maillard compound contents in the extruded blends, and to an increase (+50 mg/L) in rumen ammonia content. With reducing sugars, digestibility of Nɛ-carboxymethyl-lysine was numerically lower, and plasma EAA concentration tended to decrease, suggesting an overprotection of proteins in the intestine. With the enzymatic cocktail, plasma EAA concentration was similar than without additive during pretreatment, suggesting a similar metabolizable protein supply with both treatments. Finally, no change in N partitioning between milk, urine, and feces was observed whatever the diet.

Effects of replacing soybean meal with raw or extruded blends containing faba bean or lupin seeds on nitrogen metabolism and performance of dairy cows.

The objective was to test the effects of replacing soybean meal in dairy cow diets with either raw or extruded faba bean:linseed or lupin:linseed blends on intake, milk yield and composition, N partitioning, and ruminal and plasma parameters. Our main hypotheses were that N from extruded blends was less degradable in the rumen than N from raw seeds, and that a higher extrusion temperature favored ruminal protection of proteins and milk protein yield, and lowered urinary N excretion. Eight Holstein cows fitted with ruminal cannulas were used in two 4 × 4 Latin square design experiments conducted in parallel. In both experiments, cows were fed diets with a crude protein content of 14.6%, containing 60% of forage (dry matter basis). Treatments differed by the composition of the concentrates: control in both experiments was based on soybean meal, and experimental treatments were based on proteaginous:linseed (90:10%) blends consisting of faba bean blends (first experiment) or lupin blends (second experiment) presented either raw, extruded at 140°C, or extruded at 160°C. Intake, milk yield and composition, ruminal pH, volatile fatty acids and ammonia kinetics, digestibility, N partitioning, Maillard compounds in feed and feces, plasma AA, and 15N natural enrichment were measured. Data were analyzed using ANOVA according to the MIXED procedure of SAS (version 9.4, SAS Institute Inc., Cary, NC). Ammonia content in ruminal fluid did not significantly change when soybean meal was replaced by either raw or extruded faba bean, but tended to be higher with lupin. Milk yield was increased by 2.6 kg with faba bean blend extruded at 140°C compared with faba bean blend extruded at 160°C. Milk fat and milk protein concentrations were decreased by 3.1 and 2.3 g/kg, respectively, with lupin blends compared with soybean meal. Nitrogen partitioning between milk, feces, and urine did not change. Nitrogen apparent digestibility decreased by 3 g/100 g of N between faba bean blend extruded at low and at high temperatures. The content of Maillard compounds in feces was higher with blends extruded at 160°C than with raw or extruded at 140°C blends within both experiments. Total plasma AA tended to be higher with extruded blends than with raw in the faba bean experiment. Both extrusion temperatures appeared to protect dietary proteins from ruminal degradability, but proteins seemed to be overprotected at 160°C.