Separation、Fermentation Of Cordyceps Strains From Naqu And Evalvation Of Immunological Cordyceps Polysaccharide

Cordyceps sinensis is a precious Chinese herbal medicines and now the rare and precious resource more lack because of its unique growing conditions, coupled with excessive harvesting. This research aims at a strain of high-yield strains of Cordyceps polysaccharide expaned this precious resource.Separating fresh alpine meadows fruiting body of Cordyceps sinensis which are taken from in naqu, Tibet, got 50 strains of varying size, shape, color. Using the Insert method to observe characteristics of mycelium andconidia morphology and literaturing, found that CC-2, the CC-3, CC-4, CC-5, CC-9, CC-10 are similar to cordyceps fungus spore in morphology structure.Via biological identification, found that the similarity of CC-2 strains and Simplicillium sp is 97.80%, the similarity of CC-3 strains and Cladosporium sp is 100%, the similarity of CC-4 strains and Claviciptitaceae sp is 99.76%, the similarity of CC-5 strains and Uncultured ustilago is 99.83%, the similarity of CC-9 strains and Ustilaqinoidea virens is 100%, and the similarity of CC-10 strains and Verticillium sp is 97.1%.Selected colony white, and growth exuberant, and hyphae robust of Cordyceps strains CC-3 for fermentation medium and fermentation conditions optimization, single factors and orthogonal test results displayed, to peptone for nitrogen source, polysaccharide content for 0.686 g/L. to maltose for carbon source, polysaccharide content Supreme for 0.786 g/L. Orthogonal designoptimal proportion is KH2PO4 salts 0.1 g, Mg SO4 0.05 g, Zn SO4 0.05 g, highest content of polysaccharides 1.550g/L. Optimum fermentation conditions for the fermentation temperature is 30 degrees centigrade, inoculum 10%, the extraction rate of the highest, to 2.180 g/L.Extraction rate as an index,orthogonal water extraction and ultrasonic-assisted water extraction of Cordyceps polysaccharide extraction of optimal conditions for extraction of Hot water temperature 90 ℃, solid-liquid ratio 1:40, time 180 min, Extracted 4 times, the rate of polysaccharides to 2.265%. Ultrasonic water extraction of polysaccharides from 3:7 the optimal conditions for the start and stop time, time 90 min, 4 times ultrasound extraction ate is highes,to 6.619%. Extraction rate 2.9 times times higher than the former.Selected strains CC-3, 50 broiler as the test material.The broilerdivided into 3 groups randomly, the control group, the 1% dose group and the 2% dose group. Studies on immune organ index HE staining and the intestinal microbial flora. In the 7-14 d, the thymus grows slowly and 21 d grows rapidly. In the 28 d the thymus index and the thymus begins to degenerate. The degeneration of the 35 d was significantly lower and the degeneration of the thymus and bursa was significantly decreased after the normal capsule was degenerated. H.E slices can be seen, compared with the control group, the experimental group of the spleen and bursa of the capsule were not significantly changed, the normal tissue sections. Control group of splenic red and white pulp boundary is not obvious, the peripheral lymphatic sheath is not obvious, add Cordyceps Polysaccharide of the experimental group and the control group compared to lymphocytes is dense. The development of spleen is better than the control group. Bursal sections, control group cortex less, lymphocyte is relatively sparse, add Cordyceps Polysaccharide of test group and the control group compared to lymphocytes is dense, thick cortex and the experimental group well developed.From the test results, the number of Escherichia coli displayed significantly lower, and the difference between the experimental group and the control group was significantly(P<0.05). The number of Lactobacillus increased significantly(P<0.05), and the number of Lactobacillus increased with the increase of the amount of polysaccharide. The increase of the number of Bifidobacterium with Cordyceps polysaccharide amount, the number of bacteria in the increase of adding 1% dose group in the experimental group significantly(P < 0.05). 2% dose add group in the experimental group were significant(P < 0.01).