Differences Response Of Different Selection Lines Of Pacific Abalone,Haliotis Discus Hannai Ino To Vibrio Stimulation

Haliotis discus hannai Ino is one of the most important aquaculture products in China and it is also one of the favorite seafoods in China.Through the proliferation of breeding and breeding workers' hard work also with the popularization of various new farming modes,China has become the first abalone breeding country in the world.There are two mainly breeding species in china,one is Haliotis discus hannai another is Haliotis diversicolor,but the yield of Haliotis discus hannai has exceeded that of other countries' abalone in the world.Now the high annual production of abalone cannot be separated from the promotion of high quality abalone germplasm resource,adequate feeding of food,good quality of cultural water,careful management of the breeding and important means of disease prevention and control.Abalone is an ordinary member of mollusk.Like other marine invertebrates,there is no evolutionary antibody-specific immune system,only the non-specific immune system or the innate immune system.Abalone's innate immunity plays an important role in the complex and volatile marine environment,and the continuous improvement of innate immunity in the process of evolution with the environment makes this ancient species alive for hundreds of millions years on Earth.There are historical records of human beings has begun to use abalone before 75.000 years ago.In our country,since the ancient treasure has been used as delicious seafood.there are a lot of food related to abalone.for another thing the shell of abalone is also expensive Chinese medicine,and large-scale farming may has a relationship with this.Although there are many varieties of abalone in the world,but all kinds of abalone in the breeding process have the outbreak of some diseases,especially in large-scale intensive farming conditions,as well as environmental pollution,ocean acidification and global worming in the case of abalone breeding the marine environment is also deteriorating,which vibrio caused by the disease accounted for the vast majority.Vibrio disease is a common pathogen of marine life,which is harmful to marine life and human.Since the observation of the mortality were different after infection Vibrio,the following studies were carried out:The different tolerance of two selection lines of Haliotis discus hannai to vibrio challenge;selecting the tolerance line of abalone,and the select effect of vibrio disease resistance breeding;the different response between vibrio resistance selection line and general population of Haliotis discus hannai.Now the results of this stage are summarized as follows:1 The detection of vibrio tolerance in different strains of Haliotis discus hannai.The experimental system was tested with 3 kinds of vibrio,and the experimental period was 7 days.At different Vibrio concentrations,the mortality rate increased with the concentration of the vibrio.The tolerability of Vibrio alginolytica(bb3),"97"selection line was similar to "98" selection line,but for the tolerability of V.harveyi(bb4)and V.anguillarum(vm),98 selection line was higher than that of 97.2 Vibrio resistance line's selection and the test of screening effectR1 population was the survial abalones which used for detection of vibrio tolerance in first chapter.The final mixed vibrio concentration ofbb3,bb4 and vm was 5×106 cfu/mL,1×105 cfu/mL,and 5×106 cfu/mL to inject to 4059 health abalones.When the mortality was stable.we make a statistics that found a total survival of 321.and the cumulative mortality rate was 92.09%,in line with the general selection intensity.The selected population is defined as R2 population,and all the unselected population is defined as CK population.The mortality rate of R1 and CK population was 36.67±4.49%and 84.00±6.37%especially after the challenge of three kinds of Vibrio mixed bacteria,and the difference was extremely significant(P<0.01).We injected the same dose of bb3,bb4 or vm vibrio bacteria to R2 and CK population,which the vibrio dose were 1 X 106?1×106×3×107 cfu/mL.For bb3 injection,we found that the moratility rate of R2 population was 13.33±4.71%and CK population was 53.33 ± 9.43%.and the difference was extremely significant(P<0.01);For bb4 injection,we found that the moratility rate of R2 population was 23.33±4.71%and CK population was 83.33±12.47%,and the difference was extremely significant(P<0.01);for vm injection,we found that the moratility rate of R2 population was 0 and CK population was 36.67±4.71%,and the difference was extremely significant(P<0.01);These three tests also proved we had a successful selection.3 Different response in gene expression level between population R2 and CK against vibrio stimulationIn this study,we selected five genes that related to Haliotis discus hannai immune defense.It is Defensin,Lysozyme,HSP70,NOS and Hemocyanin.We can easy to distinguish CK and R2 population for the different expression of these genes.The results show that the relative expression of defensin gene,lysozyme gene,nitric oxide synthase gene were significantly in R2 group lower than the control group CK;but the mantle and hepatopancreas of the hemocyanin gene relative expression quantity in R2 group was higher than that of the control sample of CK;the expression of HSP70 gene of R2 group in the mantle were significantly lower than control.But the expression in gills and hepatopancreas tissues were significantly higher than the control...