Study Of Production Of Gluten Fermented By Rumen Bacteria In Vitro Improving Xueshang Chicken Laying Performance And Isolation Of Bacteria Producing Exopoid Peptides

This paper presents the results of a study on fermentation of wheat gluten protein in the artificial rumen, The study was conducted under labortatory condition. During the study invitro fermentation liquor with higher concertration of gluten peptide was extracted. The extracted liquid was found to have bio-activity of plant exorphines, and can improve Xueshan chicken lay performance and some revelent endocrine level' after hatching. Then a bacterial strain responsible for producing exopoid peptide was isolated from the fermentation mixture in vitro. This study, which comprise of four parts, privides the basis for future exploration of plant exorphines and their influence on animal performance. Presented below are the inherent series of this study.Series Ⅰ : Study on degrading gluten protein by rumen bacteriaThis experiment is to study the sweat gluten protein fermented by selected rumen bacteria in the artificial rumen, and extract the fermentation liquid with higher concertration of gluten peptide. In the whole process of fermation, fresh medium was added at the rate of 0.5ml/min in order to keep pH in 6.23-6.87. The total dehydrogenase activity(TDHA) of the bacteria was ranging from 0.65 U/ml to 1.2 U/ml, and its average level was 0.899 ± 0.014U/ml; The concertration of bacteria protein was changing from 0.306 mg/ml to 1.576 mg/ml, and its averagelevel was 0.901 ±0.041mg/mi; The average activity of proteolytic enzyme was 50.002 ±2.626U/ml, In 18h of fermentation,the concentration of gluten peptide in extracted fermentation fluid was the maximum, 253.063 ± 8.234 ug/ml.Series Ⅱ: Study on small intestine of toad in vitro to determineplant exorphins activityThe main aim of this experiment is to establish a method of determining plant exorphins activity by a small intestine in vitro. The dertermined sample is the extracted fermentation fluid in the series I experiment. In the normal state, the contraction force of small intestine in vitro was 0.230± 0.027g, After the sample was added, the contraction force increases to 0.576 ± 0.080g, which was significantly enhanced. But after the opioid peptide inhibitor-naloxone was added, the contraction force significantly decreased to 0.337 ± 0.019g. The result showed that the dertermine sample can significantly enhance the contraction force of small intestine in vitro, and this effect can be reversed by the opioid peptide special inhibitor-naloxone, which proved that the extracted fermentation fluid in the series I experiment had bio-activity of plant exorphines. Contrasting to the conventional meathods, the merits of this meathod are its fastness and convenience.Series Ⅲ Effects of degradation of wheat gluten on Laying Performance and some revelent Hormone Level in laying Xueshan chickenDifferent levels of degradation of sweat gluten with the bioactivity of plant exorphines were added to the basal diet of 20-week-old Xueshan chicken to improve lay performance and raise some revelent endocrine level after hatching. Results showed that, During 22-29weeks of age, compared with control group, treated group 3 was the most effective. The average egg weight, total egg production, laying rate and feed/egg ratio of this group were respectivelyincreased by 4.33% (P<0.01), 3.70% (p>0.05), 8.39% (P<0.01) and 2.30% (p>0.05). And the average level of serum E2, FSH, LH, P of this group was respectively increased by 25.64% (P<0.05), 16.20% (P>0.05), 24.26% (P<0.01), 2.72% (P>0.05). The results implied that appropriate level of degradation of sweat gluten with the bioactivity of plant exorphines can improve Xueshan chicken lay performance and some revelent endocrine level after hatching.Series IV Isolation of bacteria producing exopoid peptide bydegrading wheat glutenA selective medium with the wheat gluten as the only source of protein was used to islate the bacterium from the rumen of goat feeded with sweat gluten, a bacterial strain which can producing exopoid peptide by degrading sweat gluten was isolated and named OB-1. This stain is Gram-postive and has spore and h...