| With the increasing need of people for aquatic product, aquaculture gains the express development. But in the meanwhile, the water contamination, which caused by wide scale and high consistency of aquaculture, result in serious consequences of aquatic disease day by day, especially the huge loss effected by virus disease. Currently, the marine fish culturists usually use chemosynthesis or antibiotics as the therapy medicament. However, the side effects such as drug residues, resistant strains induced and water pollution etc. becomes the restriction of the development of China's aquaculture business. Compared with traditional vaccine, genetic engineering vaccine has its unique vantage, for example, low cost, ability of composition with other multivalent immunogen vaccine, inducing strong and sustained immune response, etc. Nowadays, aquaculture sector genetic vaccine research is still at an early stage, abroad study is mainly in the IHNV, VHSV, SVCV, SHRV and other infectious virus disease control of salmon and trout fish in the upper. Till now, there is no TRBIV genetic engineering vaccine study report and coverage. Here the paper filtrates two kinds antigen gene TRBIV 25R(Ribonucleotide reductase,R2/beta subunit) and TRBIV 47R(Platelet-derived and vascμlar endothelial growth factors family domain) by using bioinformatics; Builds two kinds TRBIV genetic vaccine: pVAX1 47R and pVAXl 25R, and research the expression level in grass carp kidney cells and the Xiphophorus helleri in vivo. The result shows pVAX1 47R both can express in the CKI cells and swordtail in vivo. The pVAXl 47R's expression about lasted 40 days and had a highest relative expression level in 20 days. However, the pVAXl 25R have little expression in the CKI cells and Xiphophorus helleri in vivo. For the swordtail injected pVAXl 47R, does the evaluation of immune effects. The result shows the expression of IgM, whose the swordtail injected pVAXl 47R, is significantly higher. The phenomenon indicates pVAXl 47R can stimulate the immune response in fish effectively. Study the expression time and safety of PVAX1-47 in swordtail, and further probe into its immune mechanism. The thesis provides essential theoretical basis of the production application and industrialization of TRBIV genetic engineering vaccine.In addition, the author also did the rare disease research for two marine fish species, Epinephelus septemfasciatus Thunberg and Carcharias Taurus.Epinephelus septemfasciatus Thunberg belongs to Perciformes, Serranidae, is the large kind in Epinephelus genus. It's very popular among consumers because of its tender and delicious meat. Since the demand in Japan and South Korea is great, Thunberg can be the great market potential of high-grade precious marine fish species. In the early June,2009, inflation began to appear within the eyes of some juvenile fishes of Thunbergs in a swine farm, apart from that, the juvenile fishes also had exophthalmos, swimming slowly symptoms. With the serious of the disease, corneal opacity juveniles began to appear, then blindness, eye loss, red body, not eating and other symptoms, eventually leading fish fries to death. We segregated a pathogen BY0081 from sick Thunberg, researched its pathogenicity, analyzed its physiological, biochemical and 16S rDNA sequence; after that, identified the pathogen as Pseudoalteromonas sp, then further studied its susceptibility. The study result brings positive significance for the control of fish disease which arises from Pseudoalteromonas sp.Carcharias Taurus also called Kamoharai, Sand Shark or Killer Shark Tooth Bevel Gear, is a kind of shark living in coastal waters. In Oct,2009, a Carcharias Taurus was dead accidentally in Shandong province. We searched the pathogenic factor from bacteria, parasite and virus etc. and finally identified that the cause of death is the mixed infection of Vibrio and LCDV. The outcome can contribute to the control of shark disease. |
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