| The purpose of this study was to determine the technology of extraction from prick Opuntia monacantha, study the Effects on the Newcastle disease virus in vitro, and discuss its preliminary mechanism, research duodenums mucosal immune on the chickens of the Opuntia monacantha polysaccharides.1.The optimal condition of extraction of Opuntia monacantha Polysaccharides (OMPS) by the ethanol subsiding method is established by the method of single-factor test and orthogonal experiment:the extraction temperature is 100℃,the extraction time is 3 hours, the Liquid material ratio is 1:8, and the concentration of ethanol is 70%.According to the above-mentioned conditions, the content of OMP was 38.63%, the rate of OMPS was 0.58%,the content of protein was 0.037%.2. There were three different ways:virus infection after adding OMPS, adding OMPS and Virus meantime, and adding OMPS after virus infection to be studied the effect on the Newcastle Disease Virus in Vitro of Opuntia Monacantha Polysaccharides. It was evaluated with observing the CPE of BHK cells and detecting OD450 value by the method of MTT. The results were judged by IC50, virus inhibition rate and therapeutic index. The results indicated OMPS displayed a virustatic effect in different ways. The highest inhibition ratio is the group of virus infection after adding OMPS. The higher inhibition ratio is the group of adding OMPS and Virus meantime. The worst effect is the group of adding OMPS after virus infectiont. The results show that OMPS have significant effect on inhibiting the replication of NDV, especially in the way of virus infection after adding OMPS and the concentration-response relations is evident. It indicates that preventive dose administration is the main way.3. The chickens were inoculated LaSota lived vaccine against Newcastle disease by intranasal and eye-drop route and administered to Different Doses of OMPS. The results show that the antibody titer in the serum of the chickens has greatly improved at first week after immunization. At second week after immunized, all of the antibody titer reached the peak. The differences between the high dose group and control group were very significant from the second week to the fourth week (p<0.01).The differences between model dose group and control group were very significant (p<0.01) at second week and significant (p<0.05).The quantitiy-effect relationship in the high dose, model dose and low dose are obviouse. The results indicated that the OMPS significantly (P<0.05) improved the immunization results of the ND IV vaccine, its action was of dose-dependent.OMPS can be used as immunoenhancement in practice.4. The chickens were inoculated LaSota lived vaccine against Newcastle disease by intranasal and eye-drop route and administered to Different Doses of OMPS.The content of SIgA in the intestinal mucous was measured by Enzyme Linked Immunosorbent Assay (ELISA).The positive cells and material of SIgA in duodenum and jejunum were detected by immunohistochemistry SABC method.(1)The results of the content of SIgA (secretary immunoglobulin A) in intestinal mucus were measured by ELISA.The high dose and model dose of the OMPS can obviously increase the content of the SIgA, and the doseffect relationship was obvious. At the second week, difference between the test groups and control group were all very significant (p<0.01), difference between the model dose group and low dose group were significant (p<0.05). At the third week and the fourth week, the differences between the high dose group and model dose group, the differences between the high dose group and control group, the differences between the model dose group and low group, were all very significant (p<0.01).(2)The positive cells and material of SIgA in the duodenums and jejunum were detected by immunohistochemistry SABC method.The OMPS can obviously increase the quantity of the positive cells and material of SIgA in the duodenums and jejunum, and the doseffect relationship was obvious. The changes of positive cells and material of SIgA in the duodenal and jejunal are basically in the same.At the second week, the differences in the test groups were all very significant (p<0.01), the difference between test groups and control group were all very significant (p<0.01) except low dose group. The differences between the high dose group and model dose group, and the the differences between the high dose group and control group, were very significant (p<0.01) at the third week. The the differences between the high dose group and control group in the duodenums were very significant (p<0.01), but in the jejunum were significant (p<0.05). The differents in the rest of the groups was not statistical.These results demonstrate that OMPS have increased the secretion of SIgA in the duodenums and jejunum of the chickens, when the contents of OMPS are 160mg/kg and 80mg/kg, they can significantly enhance the intestinal mucosa immune. It points out OMPS can be used as intestinal mucosa immunoenhancement in practice. |
