Study On The Degradation Rate Of Tea Saponin In Camellia Oleifera Meal And Screening And Characteristics Of Forage Probiotics

Camellia oleifera is a kind of high-value economic oil plant in south of China.Camellia oleifera meal is the residue of Camellia oleifera seed after extracting Camellia oil,and it is rich in nutrition.Tea saponin is insoluble in organic solvents.Most of the tea saponin is left in Camellia oleifera meal after extracting Camellia oil.Tea saponin has antibacterial,anti-inflammatory,stimulating hormone production,the role of elimination of snails,etc.It is a natural surfactant.However,tea saponin has some certain hemolytic property,which has a certain damage effect on animal red blood cells.At the same time,just a few tea saponin can play a beneficial role.Therefore,it is necessary to treat the tea saponin in the camellia oil meal to enable the camellia oil meal to be applied in the direction of the feed additive.At present,some methods for treating tea saponin including water extraction,solvent extraction,and auxiliary extraction.These methods consume relatively high resources and equipment for requirements,so the way of microbial degradation of tea saponin is increasingly favored by researchers.Probiotic feed additive is a new type of functional feed processing technology,which has added a little amount in feed,but play an obvious effects role on animal husbandry.As a non-toxic,non-residual and drug-resistant feed additive,probiotics have received extensive attention.How to screen the strains with high utilization rate which have the good functionality is the one of the ways to effectively use Camellia oleifera meal.Therefore,I used camellia oleifera meal as raw material to screen the microorganisms that can degrade the tea saponin in the camellia oleifera meal in the natural environment,and I optimized microbial degradation in this paper;I have screened the strain with certain probiotic function from the fresh manure of the piglet.And the strain was identified,the biological characteristics were explored,and the process of solid-state fermentation of the saponin-producing spores was studied.The main results are as follows:1.Screening of tea saponin-degrading bacteria and optimization of fermentation conditions.From the fermented camellia oil meal under natural conditions.6 strains with strong ability to degrade tea saponin were screened;strain L-2 with the best degradation effect of tea saponin was selected.The morphological identification and ITS sequence identification were carried out,and the L-2 strain was Aspergillus niger.The single factor experiment on the fermentation conditions of the tea saponin in the L-2 solid fermentation degradation camellia oil meal was carried out.The surface was optimized,and the degradation rate of tea saponin in Aspergillus niger was reduced to 93.96%.The optimum degradation conditions were fermentation temperature 31.3?,fermentation time 103.5 h,and initial acid addition amount 4.57 mL.2.Screening of probiotic strains.Making the pathogens preserved in the laboratory as the target bacteria,the CFC-11 and MR-1 with strong antagonistic ability were screened from the fresh piglet pig manure by the 80? temperature resistance method and the diluted coating plate method;It draw a conclusion from the results including identification,physiological and biochemical identification and 16S rRNA gene sequence analysis,that CFC-11 and MR-1 were identified as Bacillus licheniformis.3.The biological characteristics of probiotic strains.The growth curves of Bacillus licheniformis CFC-11 and MR-1 indicate that the two strains have short growth cycle and fast growth rate;Bacillus licheniformis CFC-11 and MR-1 for norfloxacin and gentamicin has strong sensitivity;Bacillus licheniformis CFC-11 and MR-1 can grow in a medium with a bile salt concentration of 0.1%to 0.4%;Bacillus licheniformis CFC-11 can be grown in less than 11%NaCl concentration of the medium;MR-1 can be grow in less than 10%NaCl concentration of the medium;These two Bacillus licheniformis can grow at optimum growth pH of 6;The spores of Bacillus licheniformis CFC-11 and MR-1 has certain tolerance at high temperature;We can conclude from the enzyme-producing experiments:both Bacillus licheniformis CFC-11 and MR-1 strains can produce protease and cellulase by fermentation,and we can get the stronger enzyme by the mixed fermentation.When the ratio of the two strains was 1:1,the enzyme activities of protease and cellulase reached a maximum,and the number of spores reached the maximum at this time,and the maximum value was 1.28×109 CFU/g.It follows that Bacillus licheniformis CFC-11 and MR-1 are probiotics with probiotic potential.4.Camellia oil meal fermented probiotics.Using spore number as the main evaluation index,single factor experiment and orthogonal experiment were used to optimize the mixed fermentation medium composition of Bacillus licheniformis CFC-11 and MR-1.The optimal medium composition was determined as degraded tea saponin,include 20 g of camellia oil meal,3g of glucose,0.5g of ammonium sulfate,0.25g of disodium hydrogen phosphate,18mL of deionized water,and keeping 121? for 30 min.The single-factor experiment and response surface experiment were carried out on the mixed fermentation conditions of Bacillus licheniformis CFC-11 and MR-1.The optimum fermentation temperature was 31.6? and the fermentation time was 71.1 h.The inoculum amount was 3.5%,and the number of spores reached 2.37 x 1010 CFU/g.