Rumen Degradability and Small Intestinal Digestibility of the Amino Acids in Four Protein Supplements

Wang, Y.,Jin, L.,Wen, Q.N.,Kopparapu, N.K.,Liu, J.,Liu, X.L.,Zhang, Y.G. Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.2

The supplementation of livestock feed with animal protein is a present cause for public concern, and plant protein shortages have become increasingly prominent in China. This conflict may be resolved by fully utilizing currently available sources of plant protein. We estimated the rumen degradability and the small intestinal digestibility of the amino acids (AA) in rapeseed meal (RSM), soybean meal (SBM), sunflower seed meal (SFM) and sesame meal (SSM) using the mobile nylon bag method to determine the absorbable AA content of these protein supplements as a guide towards dietary formulations for the dairy industry. Overall, this study aimed to utilize protein supplements effectively to guide dietary formulations to increase milk yield and save plant protein resources. To this end, we studied four cows with a permanent rumen fistula and duodenal T-shape fistula in a $4{\times}4$ Latin square experimental design. The results showed that the total small intestine absorbable amino acids and small intestine absorbable essential amino acids were higher in the SBM (26.34% and 13.11% dry matter [DM], respectively) than in the SFM (13.97% and 6.89% DM, respectively). The small intestine absorbable Lys contents of the SFM, SSM, RSM and SBM were 0.86%, 0.88%, 1.43%, and 2.12% (DM basis), respectively, and the absorbable Met contents of these meals were 0.28%, 1.03%, 0.52%, and 0.47% (DM basis), respectively. Among the examined food sources, the milk protein score of the SBM (0.181) was highest followed by those of the RSM (0.136), SSM (0.108) and SFM (0.106). The absorbable amino acid contents of the protein supplements accurately reflected protein availability, which is an important indicator of the balance of feed formulation. Therefore, a database detailing the absorbable AA should be established.

Measurement of the Intestinal Digestibility of Rumen Undegraded Protein Using Different Methods and Correlation Analysis

Wang, Y.,Zhang, Y.G.,Liu, Xiaolan,Kopparapu, N.K.,Xin, Hangshu,Liu, J.,Guo, Jianhua Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.10

Four methods were adopted, including the mobile nylon bag (MNB) method, modified three-step in vitro (MTS) method, original three-step in vitro (OTS) method, and acid detergent insoluble nitrogen (ADIN) estimating method, to evaluate the intestinal digestibility of rumen undegradable protein (DRUP) of 10 types of concentrates and 7 types of roughages. After correlation analysis to determine the DRUP values using the MNB, MTS, OTS, and ADIN methods, the study aimed to find out appropriate methods to replace the MNB method due to its disadvantages such as high price, long time period, and use of a duodenal T-fistula. Three dairy cows with a permanent ruminal fistula and duodenal T-fistula were used in a single-factor experimental design. The results showed that the determined DRUP values using the MNB method for soybean meal, cottonseed meal, rapeseed meal, sunflower meal, corn germ meal, corn, rice bran, barley, wheat bran, corn fiber feed, Alfalfa (Zhao dong), Alfalfa (Long mu 801), Alfalfa (Long mu 803), grass (North), Grass (Inner Mongolia), corn silage and corn straw were 98.13%, 87.37%, 88.47%, 82.60%, 75.40%, 93.23%, 69.27%, 91.27%, 72.37%, 79.03%, 66.72%, 68.64%, 73.57%, 50.47%, 51.52%, 54.05%, and 43.84%, respectively. The coefficient of determination ($R^2=0.964$) of the results between the MTS method and the MNB method was higher than that ($R^2=0.942$) between the OTS method and the MNB method. The coefficient of determination of the DRUP values of the concentrates among the in vitro method (including the MTS and OTS methods) and the MNB method was higher than that of the roughage. There was a weak correlation between the determined DRUP values in concentrates obtained from the ADIN method and those from the MNB method, and there was a significant correlation (p<0.01) between the determined DRUP values of the roughage obtained from the MNB method and those obtained from ADIN method. The DRUP values were significantly correlated with the nutritional ingredients of the feeds. The regression equation was DRUP =100.5566+0.4169CP - 0.4344SP - 0.7102NDF - 0.7950EE ($R^2=0.8668$, p<0.01; CP, crude protein; SP, soluble protein; NDF, neutral detergent fiber; EE, ether extract). It was concluded that both the MTS method and the OTS may suitable to replace the MNB method for determining the DRUP values and the former method was more effective. Only the ADIN method could be used to predict the values of the roughages but conventional nutritional ingredients were available for all of the samples' DRUP.

Measurement of the Intestinal Digestibility of Rumen Undegraded Protein Using Different Methods and Correlation Analysis

Y. Wang,Y.G. Zhang,Xiaolan Liu,N.K. Kopparapu,Hangshu Xin,J. Liu,Jianhua Guo 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.10

Four methods were adopted, including the mobile nylon bag (MNB) method, modified three-step in vitro (MTS) method, original three-step in vitro (OTS) method, and acid detergent insoluble nitrogen (ADIN) estimating method, to evaluate the intestinal digestibility of rumen undegradable protein (DRUP) of 10 types of concentrates and 7 types of roughages. After correlation analysis to determine the DRUP values using the MNB, MTS, OTS, and ADIN methods, the study aimed to find out appropriate methods to replace the MNB method due to its disadvantages such as high price, long time period, and use of a duodenal T-fistula. Three dairy cows with a permanent ruminal fistula and duodenal T-fistula were used in a single-factor experimental design. The results showed that the determined DRUP values using the MNB method for soybean meal, cottonseed meal, rapeseed meal, sunflower meal, corn germ meal, corn, rice bran, barley, wheat bran, corn fiber feed, Alfalfa (Zhao dong), Alfalfa (Long mu 801), Alfalfa (Long mu 803), grass (North), Grass (Inner Mongolia), corn silage and corn straw were 98.13%, 87.37%, 88.47%, 82.60%, 75.40%, 93.23%, 69.27%, 91.27%, 72.37%, 79.03%, 66.72%, 68.64%, 73.57%, 50.47%, 51.52%, 54.05%, and 43.84%, respectively. The coefficient of determination (R2 = 0.964) of the results between the MTS method and the MNB method was higher than that (R2 = 0.942) between the OTS method and the MNB method. The coefficient of determination of the DRUP values of the concentrates among the in vitro method (including the MTS and OTS methods) and the MNB method was higher than that of the roughage. There was a weak correlation between the determined DRUP values in concentrates obtained from the ADIN method and those from the MNB method, and there was a significant correlation (p<0.01) between the determined DRUP values of the roughage obtained from the MNB method and those obtained from ADIN method. The DRUP values were significantly correlated with the nutritional ingredients of the feeds. The regression equation was DRUP =100.5566+0.4169CP – 0.4344SP – 0.7102NDF – 0.7950EE (R2 = 0.8668, p<0.01; CP, crude protein; SP, soluble protein; NDF, neutral detergent fiber; EE, ether extract). It was concluded that both the MTS method and the OTS may suitable to replace the MNB method for determining the DRUP values and the former method was more effective. Only the ADIN method could be used to predict the values of the roughages but conventional nutritional ingredients were available for all of the samples’ DRUP.

Food Microbiology and Biotechnology : Purification and Characterization of a Novel Fibrinolytic Enzyme from Culture Supernatant of Pleurotus ostreatus

( Xiao Lan Liu ),( Xi Qun Zheng ),( Peng Zhi Qian ),( Narasimha Kumar Kopparapu ),( Yong Ping Deng ),( Masanori Nonaka ),( Naoki Harada ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.2

A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDSPAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the α and β chains of fibrinogen followed by the γ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at 45°C and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.