Effects Of Vibrio Alginolyticus Peptidoglycan On Immune Indicators, The Level Of Fatty Acid And Differentially Expressed Gene Of Litopenaeus Vannamei

The peptidoglycan was extracted from cell wall of Vibrio alginolyticus, anddissolved into physiological saline completely with the help of ultrasonic.Then the mixedsolution was injected into muscle of Litopenaeus vannamei for immunestimulation.Then it was measured about the enzyme activity of SOD and NOS,component and content of hepatopancreas fatty acid and a high throughput sequencingtranscriptome of hepatopancreas to investigate differences in gene expression afterimmune stimulation.More details followed：1. The yield rate of peptidoglycan extracted from cell wall of Vibrio alginolyticus is6.43%and the after300W30mins of ultrasonic, peptidoglycan dissolves completelyand nothing residue left.It is a new and efficient method to dissolve the peptidoglycan.2. The Vibrio alginolyticus peptidoglycan was injected into Litopenaeus vannameishrimp with different concentrations of0.05mg/mL,0.1mg/mL,0.15mg/mL.The resultsshow that nitric oxide synthase vitality (NOS), superoxide dismutase vitality (SOD) hadhighly significant difference(P<0.01).In the0.05mg/mL group, the time of NOS andSOD appeared maximum value respectively at12h and48h.In the0.1mg/mL group,NOS and SOD appeared maximum value at72h. In the0.15mg/mL group, SOD andNOS appeared maximum value also at6h.3. The same treatment as before to Litopenaeus vannamei shrimp of experimentalgroup,shrimps of control group was injected with0.1mL physiological saline.Then thehepatopancreas was tested to investigate the content and component of fatty acid.Results show that there are8kind of saturated fatty acid and C16:0and C18:0dominants; C18:1dominants of all5monoenoic fatty acid, C18:2and C20:5take a largecontent of all8polyunsaturated fatty acids. The content of saturated fatty acid declinesabout2.27%, while the largest growing content of polyunsaturated fatty acid increased at4.43%,n-3series of unsaturated fatty acids increased significantly maximizing a peak at72h after injection. n-6series of unsaturated fatty acids goes steadily and have nosignificant change. C14:0and C15:0decreases all the time even at the end of theexperiment. C16:0、C16:1and C20:0decrease overall, C22:0、C22:4、C24:1decrease at the beginning and turn to increasing. C22:4is in the undervalued at72h, while C24:1is at8h after injection.4. The same treatment as before to Litopenaeus vannamei shrimp. Analysis ofLitopenaeus vannamei transcriptome by using a high-throughput sequencing techniquewas conducted and succeeds.10,439,598,780nt bases are generated totally, In the resultsof assembly,27,873unigenes are detected, total length for unigenes is29,094,393nt,averge length is1044nt,N50is1,913nt.For functionn annotation analysis, we get14,563,8,144,13,180,11,553,6,824,7,503unigenes which annotate to the NR,NT, Swiss-Prot, KEGG, COG, GO database, respectivly, the total annotation unigenesare16,387.5. After the analysis of gene expression difference,3076genes expression quantityshow a rise after injection of peptidoglycan than control group of physiologicalsaline.While650genes decrease its expression.6. GO categories of differentially expressed genes shows that all thoses unigenedivided into49functional groups of biological-process, cellular-component andmolecular-function, such as biological adhesion, biological regulation, cellularprocess,etc.