• Asian-Australasian Journal of Animal Sciences
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• v.22 no.9
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• pp.1267-1278
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• 2009

Three beef steers fitted with permanent cannulae in the rumen and duodenum were used to determine the effects of protein supply from soyhulls (SH) and wheat bran (WB) on ruminal metabolism, blood metabolites, nitrogen metabolism, nutrient digestion and concentrations of soluble non-ammonia nitrogen (SNAN) in ruminal (RD) and omasal digesta (OD). In a 3${\times}$3 Latin square design, steers were offered rice straw and concentrates formulated either without (control) or with two brans to increase crude protein (CP) level (9 vs. 11% dietary DM for control and bran-based diets, respectively). The brans used were SH and WB that had similar CP contents but different ruminal CP degradability (52 vs. 80% CP for SH and WB, respectively) for evaluating the effects of protein degradability. Ruminal ammonia concentrations were higher for bran diets (p<0.01) than for the control, and for WB (p<0.001) compared to the SH diet. Similarly, microbial nitrogen and blood urea nitrogen were significantly increased (p<0.05) by bran and WB diets, respectively. Retained nitrogen tended (p<0.082) to be increased by SH compared with the WB diet. Intestinal and total tract CP digestion was enhanced by bran diets. In addition, bran diets tended (p<0.085) to increase intestinal starch digestion. Concentrations of SNAN fractions in RD and OD were higher (p<0.05) for bran diets than for the control, and for WB than for the SH diet. More rumendegraded protein supply resulting from a higher level and degradability of CP released from SH and WB enhanced ruminal microbial nitrogen synthesis and ruminal protein degradation. Thus, free amino acids, peptides and soluble proteins from microbial cells as well as degraded dietary protein may have contributed to increased SNAN concentrations in the rumen and, consequently, the omasum. These results indicate that protein supply from SH and WB, having a low level of protein (13 and 16%, respectively), could affect ruminal metabolism and nutrient digestion if inclusion level is relatively high (>20%).

• Animal Bioscience
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• v.35 no.10
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• pp.1592-1605
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• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• Applied Biological Chemistry
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• v.13 no.1
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• pp.29-34
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• 1970

In order to prepare digested Protein source for the Weanling Food from soybean, an attempt was made to decompose steamed soybean protein to amino acids and peptides by protease and cellulase produced from Aspergillus niger and Aspergillus sojae. In this paper, the optimum condition for digestion of soybean protein were studied and also investigated the effects of decolorization of it. As the results, followings were obtained; 1. As steaming conditions, a treatment under 15 lb of pressure and 10 minutes of heating shows most effective. 2. The optimum pH of Asp, sojae enzyme for the digestion of soybean protein is 6.0, while that of Asp. niger enzyme is 4.4. In successievly-decomposing with Asp. sojae and Asp. niger, it shows the most effective on ratio of water-soluble-nitrogen to total nitrogen and amino-nitrogen to total nitrogen than any other separate treatments. 3. The suitable amount of the enzyme solution to that of the soybean substrate paste, in volume, is 1 : 2. 4. Digestion ratio of soybean protein indicates the gradual and steady effects of increasing time of digestion, but 8 hour-digestion regarding to putrefaction was suitable. 5. The most effective decolorization was successively passed on culumns of active carbon and anion exchanger (Dowex 2-x-8) at room temperature. In separate treatments, the effective order of decolorization was as follows; (Dowex 2-x-8)>Active carborn>Amberite IR-120 6. The powder type of the soy protein source obtained by concentration below $60^{\circ}C$ contains 12.51% of moisture, 66.31% of protein, 4.25% of fat, 12.75% of carbohydrate, 4.18% of ash.

• Journal of Food Hygiene and Safety
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• v.37 no.6
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• pp.411-417
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• 2022

The purpose of this study was to evaluate protein hydrolysis and the amino acid ratio among different cuts of goat meat, such as the foreleg, hindleg, loin, and rib, using an in vitro digestion model. The corresponding cuts of beef and pork were used to compare with the goat meat. The hindleg (8.32%) and rib (8.32%) had the highest levels of protein hydrolysis among the goat cuts. There was no significant difference in protein hydrolysis between goat and pork (8.57%), ribs (P > 0.05), which had higher levels of protein hydrolysis than the beef ribs. Before digestion, the glutamine (53.44%) and glycine (11.03%) ratios were highest in the pre-digested goat foreleg and loin (P < 0.05). After in vitro digestion, goat ribs had the highest lysine ratio (17.54%) among the different cuts, and the lysine ratio was significantly higher in goat ribs than beef ribs (P < 0.05). This study provides basic data on protein hydrolysis and the amino acid composition of different cuts of goat meat, which may facilitate the evaluation of protein digestion patterns and bioavailability.

• Molecules and Cells
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• v.23 no.3
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• pp.340-348
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• 2007

Although considerable effort has been devoted in the mass spectrometric analysis of phosphorylated peptides, successful identification of multi-phosphorylated peptides in enzymatically digested protein samples still remains challenging. The ionization behavior of multi-phosphorylated peptides appears to be somewhat different from that of mono- or di-phosphorylated peptides. In this study, we demonstrate increased sensitivity of detection of multi-phosphorylated peptides of beta casein without using phosphopeptide enrichment techniques. Proteinase K digestion alone increased the detection limit of beta casein multi-phosphorylated peptides in the LC-MS analysis almost 500 fold, compared to conventional trypsin digestion (~50 pmol). In order to understand this effect, various factors affecting the ionization of phosphopeptides were investigated. Unlike ionizations of phosphopeptides with minor modifications, those of multi-phosphorylated peptides appeared to be subject to effects such as selectively suppressed ionization by more ionizable peptides and decreased ionization efficiency by multi-phosphorylation. The enhanced detection limit of multi-phosphorylated peptides resulting from proteinase K digestion was validated using a complex protein sample, namely a lysate of HEK 293 cells. Compared to trypsin digestion, the numbers of phosphopeptides identified and modification sites per peptide were noticeably increased by proteinase K digestion. Non-specific proteases such as proteinase K and elastase have been used in the past to increase detection of phosphorylation sites but the effectiveness of proteinase K digestion for multi-phosphorylated peptides has not been reported.

• Current Research on Agriculture and Life Sciences
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• v.31 no.2
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• pp.108-112
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• 2013

Nuts are one of the most common sources of allergies in individuals of all ages. In order for a particular protein to render an allergic reaction, it must resist proteolytic digestion by intestinal enzymes. In this study, three well-known allergenic nuts, almonds, cashew nuts, and peanuts, were used as samples, and enzyme digestion with Bacillus protease and porcine pepsin was tested. A proteomic approach using two-dimensional gel electrophoresis and an MS/MS analysis was applied to visualize and identify the proteins that were resistant to enzyme digestion. Among the 150 protein spots tested, 42 proteins were assigned functions. Due to the lack of genomic databases, 41% of the identified proteins were grouped as hypothetical. However, 12% of them were well-known allergens, including AraH. The remainder were grouped as storage, enzymes, and binding proteins.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.12
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• pp.1849-1857
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• 2007

The experiment was carried out using fistulated multiparous Holstein Friesian crossbred (75% Holstein Friesian and 25% Red Sindhi) dairy cows in their dry period fed on untreated rice straw to evaluate the nutritive value of local protein feed resources using the in sacco method and in vitro pepsin-pancreatin digestion. Experimental feeds were cottonseed meal (CSM); soybean meal (SBM); dried brewery's grains (DBG); palm kernel meal (PSM); cassava hay (CH); leucaena leaf meal (LLM). Each feedstuff was weighed into duplicate nylon bags and incubated in each of the two rumen fistulated cows for 0, 2, 4, 8, 16, 24, and 48 h. Rumen feed residues from bags of 16 h incubation were used for estimation of lower gut digestibility by the technique of in vitro pepsin-pancreatin digestion. Ruminal ammonia-nitrogen ($NH_3-N$) concentrations did not differ between treatments or time with a mean of 5.5 mg%. Effective degradability of DM of CSM, SBM, DBG, PSM, CH and LLM were 41.9, 56.1, 30.8, 47.0, 41.1 and 47.5%, respectively. Effective degradabilities of the CP in feedstuffs were 49.6, 59.2, 40.9, 33.5, 47.3 and 65.0% for the respective feedstuffs. The CP in vitro pepsin-pancreatin digestibility as ranked from the highest to the lowest were SBM, CSM, LLM, CH, DBG, PSM, respectively. The intestinal and total tract digestion of feedstuffs in the current study were relatively lower than that obtained from previous literature. The results of this study indicate that SBM and LLM were highly degradable in the rumen, while CH, CSM and DBG were less degradable and, hence resulted in higher rumen undegradable protein. Soybean meal and LLM could be used to improve rumen ecology whilst CH, CSM and DBG could be used as rumen by-pass protein for ruminant feeding in the tropics.

• Journal of Aquaculture
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• v.5 no.1
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• pp.1-7
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• 1992

Digestion rates of protein, carbohydrate, and lipid in the two kinds of extruding processed soybean meals, heated (H-SBM) and raw (R-SBM), were tested for evaluate the effectiveness of soybean meal in the rainbow trout diet. The relation between digestion rate of protein and trypsin inhibitor (TI) was also determined. The protein digestion rate of both H-SBM and R-SBM were increased up to $95{\%}$ by extruding process compared to the none treated soybean meal. The digestion rate of carbohydrate in R-SBM was increased by extruding process whereas the one in H-SBM was not. The activity of trypsin inhibitor was almost diminished by the extruding process and digestion rate of dietary protein was improved. However, the reason of this improvement was not clear whether caused by the deactivation of trypsin inhibitor.

• Journal of Animal Science and Technology
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• v.66 no.2
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• pp.374-386
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• 2024

To predict the apparent total tract digestibility (ATTD) of crude protein (CP) in dogs we developed an in vitro system using an in vitro digestion method and a statistical analysis. The experimental diets used chicken meat powder as the protein source, with CP levels of 20% (22.01%, analyzed CP value as dry-based), 30% (31.35%, analyzed CP value as dry-based), and 40% (41.34%, analyzed CP value as dry-based). To simulate in vivo digestive processes a static in vitro digestion was performed in two steps; stomach and small intestine. To analyze ATTD the total fecal samples were collected in eight neutered beagle dogs during the experimental period. CP digestibility was calculated by measuring CP levels in dog food, in vitro undigested fraction, and dog feces. In result, CP digestibility at both in vivo and in vitro was increased with increasing dietary CP levels. To estimate in vivo digestibility the co-relation of in vivo ATTD and in vitro digestibility was investigated statistically and a regression equation was developed to predict the CP ATTD (% = 2.5405 × in vitro CP digestibility (%) + + 151.8). The regression equation was evaluated its feasibility by using a commercial diet. The predicted CP digestibility which was calculated by the regression equation showed high index of similarity (100.16%) with that of in vivo in dogs. With that, it would be a feasible non-animal method to predict in vivo CP digestibility by using in vitro digestion method and the proposed linear regression equation in adult dogs.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.2
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• pp.187-193
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• 2014

As a result of the cost of grains, the replacement of grains by co-products (i.e. DDGS) in feedlot diets is a common practice. This change produces diets that contain a lower amount of starch and greater amount of fibre. Hypothetically, combining feed grade urea (U) with slow release urea (Optigen) in this type of diet should elicit a better synchrony between starch (high-rate of digestion) and fibre (low-rate of digestion) promoting a better microbial protein synthesis and ruminal digestion with increasing the digestible energy of the diet. Four cannulated Holstein steers ($213{\pm}4$ kg) were used in a $4{\times}4$ Latin square design to examine the combination of Optigen and U in a finishing diet containing different starch:acid detergent fibre ratios (S:F) on the characteristics of digestive function. Three S:F ratios (3.0, 4.5, and 6.0) were tested using a combination of U (0.80%) and Optigen (1.0%). Additionally, a treatment of 4.5 S:F ratio with urea (0.80% in ration) as the sole source of non-protein nitrogen was used to compare the effect of urea combination at same S:F ratio. The S:F ratio of the diet was manipulated by replacing the corn grain by dried distillers grain with solubles and roughage. Urea combination did not affect ruminal pH. The S:F ratio did not affect ruminal pH at 0 and 2 h post-feeding but, at 4 and 6 h, the ruminal pH decreased as the S:F ratio increased (linear, p<0.05). Ruminal digestion of OM, starch and feed N were not affected by urea combination or S:F ratio. The urea combination did not affect ADF ruminal digestion. ADF ruminal digestion decreased linearly (p = 0.02) as the S:F ratio increased. Compared to the urea treatment (p<0.05) and within the urea combination treatment (quadratic, p<0.01), the flow of microbial nitrogen (MN) to the small intestine and ruminal microbial efficiency were greater for the urea combination at a S:F ratio of 4.5. Irrespective of the S:F ratio, the urea combination improved (2.8%, p = 0.02) postruminal N digestion. As S:F ratio increased, OM digestion increased, but ADF total tract digestion decreased. The combination of urea at 4.5 S:F improved (2%, p = 0.04) the digestible energy (DE) more than expected. Combining urea and Optigen resulted in positive effects on the MN flow and DE of the diet, but apparently these advantages are observed only when there is a certain proportion of starch:ADF in the diet.