Study On Extraction By PEF And Enzymatic Method, Bacteriostasis And Immunocompetence Of Crude Polysaccharides In Abalones' Viscus

In recent years, with the continuous improvement and innovation of the scientific level, theliving standards of people have been greatly improved. At home and abroad, the abaloneaquaculture industry has also had a rapid development. More and more people can afford theabalone. With the expansion of consumer market of the abalone, abalone products have beenincreasingly diversified, which causes a lot of abalones' viscus. Abalones' viscera contain richnutrients such as polysaccharides, proteins, amino acids and trace elements.In this paper, theabalones' viscus were the experimental materials extracted by the method of PEF combined withthe enzymolysis. On the basis of our processing, the optimal enzyme solution process conditionswere determined. Simultaneously, several components in dry powder of crude polysaccharidesextracted from abalones' organs were determined. What's more, some preliminary researches onthe functionality of the crude polysaccharides were conducted. The inhibitory effect of crudepolysaccharides in abalones' viscus on several common pathogens and its effects of immunefunction on normal mice and the mice with immune suppression were studied. The results werelisted as follows:1. The influence of PEF on extracting crude polysaccharides in abalones' viscus was studied.The result showed that the extraction effect was the best when the electric intensity was30KV/cm, and pulse number was12.2. The effects of three kinds of proteases on extracting crude polysaccharides in abalones'viscera were compared. It turned out that the neutral protease was the most optimal proteases.3. The optimum technological condition was the combination of PEF and neutralprotease.The best parameters in enzymolysis were listed as follows: the ratio of raw material towater was1:6, enzymolysis time was2.5h, the quantity of enzyme was2.4×104U/g, and thetemperature was55℃.4. Several components in crude polysaccharides powder of the abalones' viscus weredetermined. It indicated that crude polysaccharides powder contained60.39%polysaccharides,10.94%moisture,6.02%fat,17.88%protein, and0.40mg/Kg heavy metal cadmium.5. The circular filter paper method was used to study the inhibition of crude polysaccharidesin abalones' viscus on11kinds of pathogens. The results showed that the crude polysaccharidesplayed an obviously suppressive role on Salmonella typhimurium, Bacillus subtilis, Ralstonia solanacearum, Escherichia coli, and Staphylococcus aureus.But for Saccharomyces cerevisiae,Fusarium oxysporum f. sp. Vasinfectum, Penicillium, Penicillium digitatum, Fusariumgraminearum Schw, Pyricularia oryzae Cav, they had no apparent inhibition.6. The dilution-2method was used to study minimum inhibitory concentrations of crudepolysaccharides in abalones' viscus on5kinds of bacteria. The results proved when theconcentration of the crude polysaccharides in abalones' viscus was50mg/ml, Staphylococcusaureus, Ralstonia solanacearum, Escherichia coli, Bacillus subtilis could be significantlyinhibited. However, the MIC was12.5mg/mL for Salmonella typhimurium.7. Through administration of drug in vivo and setting the blank control group, low, mediumand high dose (200,400,600mg/Kg) groups of crude polysaccharides in abalones' viscus,theimpact of crude polysaccharides on the index of the organs of normal mice (including the spleenindex and thymus index), DTH, and the WBC were examined. The experimental results showedthat there weren't obvious influence of the abalones' viscus polysaccharides on the above threeindices of the mice immune function.8. Through administration of drug in vivo and setting the blank control group,cyclophosphamide model group, low, medium and high dose (200,400,600mg/Kg) groups ofcrude polysaccharides in abalones' viscus respectively with cyclophosphamide treatment group,the impact of crude polysaccharides on the index of the organs of immunosuppressed mice(including the spleen index and thymus index), DTH, WBC, carbon clearance ability wereexamined. The experimental results revealed that crude polysaccharides in abalones' viscuscould significantly improve the organs index of immunosuppressed mice (including the spleenindex and thymus index), DTH, WBC counts, carbon clearance ability. The indices werestatistical significance (P <0.05or P <0.01) by comparing the results of those indices with themodel group.And the results showed that the crude polysaccharides could enhance theimmunologic function on the above four indices.