A Study On The In Vitro Enzyme Method To Evaluate The Quality Of Phytase And NSP Enzymes

With the development of livestock industry and feed industry,enzymes were widely used as feed additive.Phytase can increase availability of phytate-P,reduce add inorganic phosphorus to poultry feed,decrease the pollution of environment.NSP enzyme can hydrolyze NSP and reduce the intestinal viscosity,which improve feed conversion.Now,enzyme has difference in production strains,gene source and the way of use,meanwhile,it has problem in storage conditions and time,so the activity evaluation method lack of uniform standard,the real effect was not objective evaluation in the practical production.The objects of this study were to establish a method imitating broiler digestion in vitro and test its feasibility and validity in digestion parameters and animal trial.Study of phytase digestion parameters in vitro.According to the results of preliminary studies,focus on the trypsin digestion time.Choosing corn sample,three treatments as add two phytase and not add phytase(control).pepsin digestion 90 min,trypsin digestion2 h,4h,6h,8h.After digestion,determine the content of P.The result showed that,at the time of2,the release of P was the highest.Combined with previous research results,we build digestive system parameters assessed phytase product quality through a two-step enzyme.Study of phytase product quality evaluation method in vitro.We collected eight kinds of phytase(A-H),ues the method to evaluated invitro.We chosen six phytase sample,placed the sample at the average temperature 33? and the average humidity 55%.The sample were collected at the first,3th,7th,14 th day and use the vitro digestion to evaluate them.The result showed that,the content of Phosphorus release increased significantly by add phytase(P<0.05).Different phytase had significant difference(P<0.05).The activity of phytase C was high,the activity of phytase B was low.Under the hot and humid condition,the activity of enzyme had decreased.the release of P obviously decreased along with the prolonged(P<0.05).So,we choose high activity of phytase C and low activity phytase B for animal trial.The production validation of animal trial.we choose high activity of phytase C and lowactivity phytase B for animal trial.We choose 640 7-day-old Arbor Acres(AA),birds were grouped randomly into 4 treatments,with 4 replicates per treatment and 40 birds per replicate.Four diets were prepared: control group feed basic ration,experimental group feed low P basic ration(available phosphorus reduce 0.2%),low P basic ration+phytase B,low P basic ration+phytase C.The apparent digestibility of phosphorus of feed were obtained by metabolism trials.The result showed that,after add phytase,.the body weigh and body weigh gain were higher than low P treatment(P<0.05).Phytase supplementation had significant increased apparent digestibility of phosphorus(P<0.05).Supplemented phytase into low P diets had increased the concentration of serum P.At 42 days of age,phytase C treatment P level reach to control treatment,higher than low P and phytase C treatment(P<0.05).phytase significant improved the calcium and phosphorus content of tibia and femur(P<0.05).Phytase C had significant higher than phytase B(P<0.05).Tests showed that phytase C was superior to phytase B.The result show,Two different qualities phytase products have an impact on growth and bone deposition of calcium and phosphorus in broiler test,in accord with the result of in vitro digestion.Study of NSP enzyme evaluation method in vitro.we have established the technology of in vitro two-step enzymatic method of pepsin-pancreatin.We evaluated the kinds of eight NSP enzyme(A-H).The result showed that,add NSP enzyme had no effect in vitro dry matter digestion.After enzyme digestion the content of NSP had reduced(P<0.05).different NSP enzyme had different effect(P<0.05).The activity of NSP enzyme H was high,the activity ofNSP enzyme B was low.we choose high activity of phytase H and low activity phytase B for animal trial.The application validation of the quality of NSP enzyme.we choose high activity of phytase H and low activity phytase B for animal trial.We choose 288 1-day-old Arbor Acres(AA),birds were grouped randomly into 4 treatments,with 6 replicates per treatment and 12 birds per replicate.Four diets were prepared:control feedwheat-soybean diet,experimental group feed wheat-soybean diet,wheat-soybean diet+NSP enzyme B,wheat-soybean diet+NSP enzyme H.The result showed that Wheat-soybean diet had effect on body weigh?feed intake and feed conversion ratio from 1 to 14 days of age(P<0.05).But the feed intake and body weigh had no effect in the whole growth period(P>0.05).AddingNSP enzyme had no effecct on feed intake and body weigh.Adding NSP enzyme improved the concentration of serum glucose(P<0.05).But the concentration of serum uric acid and AME were not significance.Compare with four treatments,except ileum length had difference(P<0.05),others had no significant effect(P>0.05).In summary,the digestion system parameter optimization in vitro,we constructed pepsin-trypsin digestion step assessment of phytase and NSP enzyme product quality methods in vitro.We conducted sample verification and production test verification,evaluation method more reliable,especially phytase assessment methods to verify the results were in agreement,to meet the production needs assessment.