Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media

Acrylamide (CH2CHCONH2), an industrially produced α,β-unsaturated (conjugated) reactive molecule, is used worldwide to synthesize polyacrylamide. Polyacrylamide has found numerous applications as a soil conditioner, in wastewater treatment, in the cosmetic, paper, and textile industries, and in the laboratory as a solid support for the separation of proteins by electrophoresis. Because of the potential of exposure to acrylamide, effects of acrylamide in cells, tissues, animals, and humans have been extensively studied. Reports that acrylamide is present in foods formed during their processing under conditions that also induce the formation of Maillard browning products heightened interest in the chemistry, biochemistry, and safety of this vinyl compound. Because exposure of humans to acrylamide can come from both external sources and the diet, a need exists to develop a better understanding of its formation and distribution in food and its role in human health. To contribute to this effort, this integrate...

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Chemistry, biochemistry, and safety of acrylamide. A review.

Soy protein is a major component of the diet of food-producing animals and is increasingly important in the human diet. However, soy protein is not an ideal protein because it is deficient in the essential amino acid methionine. Methionine supplementation benefits soy infant formulas, but apparently not food intended for adults with an adequate nitrogen intake. Soy protein content of another essential amino acid, lysine, although higher than that of wheat proteins, is still lower than that of the milk protein casein. Adverse nutritional and other effects following consumption of raw soybean meal have been attributed to the presence of endogenous inhibitors of digestive enzymes and lectins and to poor digestibility. To improve the nutritional quality of soy foods, inhibitors and lectins are generally inactivated by heat treatment or eliminated by fractionation during food processing. Although lectins are heat-labile, the inhibitors are more heat-stable than the lectins. Most commercially heated meals retain up to 20% of the Bowman-Birk (BBI) inhibitor of chymotrypsin and trypsin and the Kunitz inhibitor of trypsin (KTI). To enhance the value of soybeans in human nutrition and health, a better understanding is needed of the factors that impact the nutrition and health-promoting aspects of soy proteins. This paper discusses the composition in relation to properties of soy proteins. Also described are possible beneficial and adverse effects of soy-containing diets. The former include soy-induced lowering of cholesterol, anticarcinogenic effects of BBI, and protective effects against obesity, diabetes, irritants of the digestive tract, bone, and kidney diseases, whereas the latter include poor digestibility and allergy to soy proteins. Approaches to reduce the concentration of soybean inhibitors by rearrangement of protein disulfide bonds, immunoassays of inhibitors in processed soy foods and soybean germplasm, the roles of phytoestrogenic isoflavones and lectins, and research needs in all of these areas are also discussed. This integrated overview of the widely scattered literature emphasizes general concepts based on our own studies as well as recent studies by others. It is intended to stimulate interest in further research to optimize beneficial effects of soy proteins. The payoff will be healthier humans and improved animal feeds.

Nutritional and Health Benefits of Soy Proteins

Chemical factors involved in ruminal fiber digestion.

https://agrinews.es/wp-content/uploads/2014/01/Essential-Oils-as-Modifiers-of-Rumen-Microbial-Fermentation-S.Calsamiglia-2007.pdf

Invited review: Essential oils as modifiers of rumen microbial fermentation.

Effects of incremental urea supplementation on ruminal ammonia concentration and bacterial protein formation.

In vitro procedures for estimating rates of ruminal protein degradation and proportions of protein escaping the rumen undegraded.

The effect of heat treatment of ruminal protein degradation and escape was studied using in vitro incubations with autoclaved and commercial cottonseed meal (CSM) samples. Incubations using high ratios of protein to ruminal fluid appeared to overestimate ruminal degradation of CSM protein relative to casein. A biexponential model, assuming CSM contained two protein fractions degraded at two different rates, was used to interpret data from in vitro incubations conducted using ratios of protein to ruminal fluid similar to those expected in vivo. The first fraction was degraded at rates (0.68-1.19/hour) which were 2--3 1/2 times greater than that of casein (0.34/hour). Degradation rates of the second fraction were much slower (0.011-0.093/hour). The effect of heat treatment was to decrease the proportion of the rapidly degraded fraction and to both increase the proportion and to decrease the degradation rate of the more slowly degraded fraction. Estimated ruminal escape increased with each increment of heat treatment. Intestinal protein digestibility (ruminal escape times true digestibility) increased to maximum at 60 minutes autoclaving, and then declined. Estimates of intestinal protein digestibility averaged 30.6 and 50.3% for solvent-extracted and screw-press CSM, respectively. These and previous results suggest heat treatment decreases ruminal degradation partly by blocking reactive sites for microbial proteolytic enzymes and partly by reducing protein solubility.

Effect of Heat Treatment on Ruminal Degradation and Escape, and Intestinal Digestibility of Cottonseed Meal Protein

Chemical Inhibition of Amino Acid Deamination by Ruminal Microbes in Vitro

Effects of heating cottonseed meal (CSM) protein were quantitatively assessed by determination of true digestibility (TD), in vitro proteolysis, N solubility and fluorodinitrobenzene (FDNB) available lysine. Flaked, dehulled cottonseed was extracted with hexane and desolventized at 25 C, then autoclaved (121 C, 1.1 kg/cm(2)) for 0, 15, 30, 60, 90 or 120 minutes. Free gossypol was subsequently extracted, and TD was determined in weanling rats. Metabolic fecal N (the fecal N excreted by rats fed a basal diet containing 4% casein protein) was 1.84 +/- .10 mg N/g dry matter intake. TD and FDNB-available lysine (percentage of total) were 91 and 89%, respectively, in the unheated meal. TD and FDNB-available lysine were reduced to 84 and 78% after 60 min of autoclaving, and to 71 and 44% after 120 min of autoclaving. The effect of heat treatment on TD was described by the equation: % TD = 100 - 9.28e(.0096t) (r = .998), where t = minutes of autoclaving. This indicated an accelerated decline in TD as heating time increased. No more than 40% of the loss in FDNB-available lysine was attributable to gossypol binding. In vitro release of total amino acids from autoclaved CSM samples during pepsin-pancreatin incubations was highly correlated to TD (r = .996), but N solubility in .02 N NaOH was poorly correlated to TD. In samples of solvent-extracted and screw-pressed CSM, TD (estimated from pepsin-pancreatin incubations) ranged from 80 to 85% and FDNB-available lysine ranged from 73 to 85%, and both were only slightly lower in screw-pressed than in solvent-extracted meals. Intake of FDNB-available lysine was correlated (r = .902) to weight gain in rats fed diets containing the CSM that were more severely autoclaved. Results suggest that heat treatment must be more severe than that which normally occurs in commercial CSM processing to cause substantial, selective loss in lysine availability.

Glen A. Broderick

Papers

Effects of incremental urea supplementation on ruminal ammonia concentration and bacterial protein formation.

In vitro procedures for estimating rates of ruminal protein degradation and proportions of protein escaping the rumen undegraded.

Effect of Heat Treatment on Ruminal Degradation and Escape, and Intestinal Digestibility of Cottonseed Meal Protein

Chemical Inhibition of Amino Acid Deamination by Ruminal Microbes in Vitro

Effect of heat treatment on true digestibility in the rat, in vitro proteolysis and available lysine content of cottonseed meal protein.