The Application Research On Effect Of Solid Fermentation Phytase And Lysozyme To Broiler And Piglet

This study carried out the industrial production experiment on Aspergillus niger HF-2008 transgenic engineering bacteria and lysoxyme to produce the phytase and lysozyme that met the feed industry application. Particles and liquid phytase were produced after liquid coating and preservation experiment. In order to investigate the effects of phytase and lysozyme on the production performance of broilers and piglets, the following experiment were carried out: industrial production experiment on phytase solid fermentation; liquid phytase preservation and particles phytase heat-resistant experiment; experiment on the effects of production performance of broilers fed with corn-soybean basal diets adding phytase; experiment on the effects of the growth performance and serum biochemical parameters of weanling piglets fed with granulate diet adding aniso-form phytase, experiment on synthetic canine lysozyme gene prokaryotic expression and activity analysis, experiment on the performance of lysozyme, metabolism and immune function of broilers fed with corn-soybean basal diets, the results show:1. By the solid-state industrialization fermentation of Aspergillus niger HF-2008 transgenic engineering bacteria, the best fermentation medium were bran (adding modest powder together) 2700 kg, bean powder 400 kg, ammonium sulfate 40 kg, K2HPO4 20 kg. The best fermentation time was 70 h and the best temperature was the previous 30℃, later adjusted to 28℃. pH was 5.0～5.5.2. The best preservation of the liquid preservative was acetic acid, sodium benzoate, tween 20 (80) respectively. The ratio was 0.02%, 0.02%, 0.01% and 0.1%, respectively. Enzyme activity preservation rate of the particle phytase was 83%at 85℃.3. Dealed with two–factor variation, 400 one-day-old broilers were randomly divided into 10 groups and each group contained four replications and 10 chicken per replication (5 males and 5 females). This experiment was to investigate the effects of different dosage-form phytase (liquid, powdery and granular) and pelleting temperature (75℃, 80℃and 85℃) on growth performance, metabolism, blood biochemistry target and quality of tibia bone of broilers fed low phosphorus corn-soybean diets. Earlier period(1～21 d), the F/G of adding granular phytase group was significantly lower than adding powdery phytase group (P<0.05). Later period (22～42 d) and entire period (1～42 d), the average daily gain(ADG) of adding granular phytase group was significantly higher than adding powdery phytase group (P<0.05). The F/G was significantly lower than adding powdery phytase group (P<0.05). Entire period, the ADG of 75℃pelleting group was significantly higher than 85℃pelleting group (P<0.05), and the F/G was significantly lower than 85℃pelleting group (P<0.05). The apparent digestibility of phosphorus of each experimental group broilers was significantly improved (P<0.05). The P excretion of each experimental group broilers was significantly reduced (P<0.05). The shear force of tibia bone of adding granular phytase group was significantly higher than adding powdery phytase group (P<0.05). The serum P of 75℃pelleting group was significantly higher than 85℃pelleting group (P<0.05). The serum AKP, HYP was significantly lower than 85℃pelleting group (P<0.05). 42 age, the shear force, ash of tibia bone of adding granular phytase group was significantly higher than adding powdery phytase group (P<0.05). The phosphorus, shear force, ash of tibia bone of 75℃pelleting group were significantly higher than 85℃pelleting group (P<0.05).4. A total of 224 crossbred piglets (Duroc×Landrace×Large white), weight anaverage (11.79±0.26) kg were randomly divided into 4 groups, consisting of replicates of 14 pigs each. The control group fed the corn-bean of 83℃granulate diet. In the experiment groups (A. B. C) , 7.0 kg calcium phosphoric dibasic was replaced by 100 g or 100 ml phytase (Powdery, Pellet,liquid,500 unit), and adding 1.5kg stone powder and 5.5 kg carrier to per ton holo-valence power, granulating after sufficiently mixing. The experiment lasted 30 days. The results showed: under 83℃granulation temperature, granulation and reducing the level of dietary calcium hydrogen phosphate with adding particles and liquid phytase could significantly improve the ADG , significantly lower the F/G and also impact the piglets blood TP, ALB and GLO index (P<0.05).5. The recombinant expression vector pPROHTa-rcLYZ was constructed successfully. After the expression vector was induced at optional temperature, there was a proper band at the 16 ku. On the basis of synthetic canine lysozyme gene (396 bp, 132 aa), the lysozyme activities analysis revealed that: rcLYZ showed powerful lysozyme activity on some bacterial, e.g. Escherichia coli., icroccusly sodeikticus, and had restriction activity on some bacterials. Moreover, it was against some bacterial. The animal experiment indicated: the recombinant protein contributed to healing of artificial infection of canine E.coli disease.6. The effects of exogenous enzymes on the performance, metabolism and immune function in broilers were investigated in this paper. A total of 288 1-day-old broilers were randomly divided into 4 groups, 6 repeats in each group and 12 chickens in each repeat (6 males and 6 females). GroupⅠwas control fed with corn-soybean basal diet, but groupⅡ,ⅢandⅣwere fed with basal diet supplemented low, middle and high level of exogenous enzymes respectively. The results showed that: supplementation of exogenous enzymes decreased the feed intake, weight gain and improved feed conversion rate in broilers. In 0-6 weeks, feed intake of groupⅣwas decreased by 5.85%(P<0.05) and feed/gain was improved by 8.25%(P<0.01)comparing control respectively; supplementation of exogenous enzymes improved the indexes of immune organs, in 0～3 weeks, fabricius comparative weight of groupⅣwas increased by 27.69%(P<0.01)and thymus comparative weight was increased 14.24%(P<0.05)comparing control respectively, in 4～6 weeks, fabricius comparative weight of groupⅣwas increased 25.68%(P<0.05)and thymus comparative weight was increased 29.01%(P<0.01)comparing control respectively. Supplementation of exogenous enzymes increased the contents of TP, ALB and GLOB in serum and blood of broilers. In 0～3 weeks, TP of groupⅢwas increased by 17.58%(P<0.01)and GLOB was increased by 21.86%(P<0.01)comparing control respectively. Apparent metabolic rate of energy and nutrients, the content of IgG in serum and content of HI in blood also increased, but not significantly(P>0.05).