| PRRSV is known as porcine reproductive and respiratory syndrome (pocrin reoductive and respiratory syndrome, PRRS) is caused by blue ear pig virus an pregnancy sow abortion, various age pigs (especially the piglet) breathing disorder and the highly contagious diseases characterized by a large number of death [1]. PRRSV in pig alveolar macrophages (PAM) in a large number of replication, cause the body's immune adjustment disorder, secondary infection caused by other diseases. At present our country mainly through vaccine immunization to prevent PRRS, blue ear vaccines are now on the market basically has two kinds, one kind of PRRS inactivated seedlings; A weak for PRRS poisonous plants (classic strain or highly pathogenic strains). Weak poison immune effect is good, so are more widely used, but weak poison seedlings scattered poison itself exists dangerous[2],the pigs exist residual poison. Therefore seek to be able to use the immune effect is good as well as a way of weak blue ear poison plants and to a certain extent, reduce and purify the weak blue ear poison plants in pigs of residual poison method is particularly important.1 astragalus polysaccharides powder and radix isatidis granule on macrophage phagocytosis of mice.Objective:to study the different dosage of radix isatidis, astragalus polysaccharides powder particles on the mice immune function of macrophage phagocytosis. Methods: selected 27 mice were randomly divided into nine groups, each group of three mice. By intraperitoneal injection of 1-8 groups of mice respectively 4 different concentrations of radix isatidis particle, astragalus polysaccharides powder solution, the control injection of saline,1 times a day,4 d, consecutive day 8 intraperitoneal injection of soluble starch. produce different source sex, induced macrophage,10 d injection of 1 ml chicken reel blood cells,5% free mice activities after 30 min executed mice abdominal cavity fluid on the slide,37? temperature box after incubation in red staining microscopy, determine the change of the function of macrophage cell in abdominal cavity. Results:two drugs both can enhance the phagocytosis of macrophages in mice; Radix isatidis particle in 20 mg/ml, astragalus polysaccharides powder in 30 mg/ml works best in mice, and extremely significant difference in the control group (P<0.01).2 fluorescence quantitative rt-pcr detection of PRRSV nucleic acid residual poison methodThis experiment through the contrast analysis of PRRSV ORF7 gene sequences[3],in its conservative region design 1 of primers to artificial synthesis of target genes, after amplification of target genes length of 162 bp, the stripe is clear, the desired amplification effect. Target genes of DNA recovery, by connecting transformation, the preparation of recombinant plasmid pMD19-T-PRRSV, preliminary identification of using polymerase chain reaction (PCR), in determining the target genes into DH5 alpha success after sequencing appraisal, and the sequencing results and login on NCBI PRRSV ORF7 gene homology of 100%, that cloned target genes, and the preparation of recombinant plasmid extraction after determination of the concentration as a fluorescence quantitative rt-pcr amplification of the template. According to the target gene sequence design degenerate primers for fluorescence quantitative PCR amplification. At the same time through optimizing degenerate primer concentrations and annealing temperature to determine the method of the optimum reaction conditions, successfully established on PRRSV SYBR Green I fluorescent quantitative PCR detection method, the design of the degenerate primers easier for strain identification between different regions. Through the specificity, sensitivity and repeatability test, the method can specificity of detected pMD19-T-PRRSV, Yang of plasmid template can be detected low copy number of 1.66×101 copies, Yang plasmid template within the group are less than 2%. variation coefficient of show that the method has good specificity, high sensitivity. Late for drug testing PRRSV nucleic acid load variation in monitoring the pig blood laid a foundation.3 astragalus polysaccharides powder and radix isatidis particle reduce blood PRRSV residual poison poison study (or vaccine)Purpose:by astragalus polysaccharides powder and radix isatidis particle feeding, detecting poisonous sows PRRSV residual poison vaccine (or drug) in the blood, astragalus polysaccharides powder was revealed and radix isatidis particle decrease in pig blood PRRS residual poison vaccine (or drug) ability, provide a reference for production actually guarantee the herd of swine health and help facilitate astragalus polysaccharides medicinal powder and radix isatidis particle in the development and application of additives. Selection methods:21 first tested with residual poison PRRS vaccine (or drug) of the gilt, randomly divided into three groups, each join in each group of three head without toxic health sows, namely the control group and experiment 1 group,2 group. Control group fed chow, with 1 set of astragalus polysaccharides powder mixing materials feeding, use of radix isatidis particle mixing materials feeding experiment 2 group. Three groups of test sows fed separately, feeding cycle for two months, once every ten days for blood through the establishment of fluorescence quantitative rt-pcr technique to detect the sow in the blood the PRRS residual poison the nucleic acid vaccine (or drug) loads. The results show that the two drugs can protect health sows from PRRSV infection to some extent. And for the sow has been poisoned, radix isatidis particle can effectively reduce the PRRSV loads of nucleic acid in the blood, help the sow purification of PRRSV residual poison in the blood, decrease the rate of sow the elimination and returned to the feeling, and the effect of astragalus polysaccharides powder is not obvious. |
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