Research Into The Easing Of Gln To Immunological Stress Mechanism In Trionyx Sinensis

Trionyx sinensis,an old secondary aquatic reptiles,are of research importance. Since 1980s,the increasing market demand has stimulated the rapid development of Trionyx sinensis breeding.Trionyx sinensis is always under the state of stress under the impact of various kinds of environment,human management and pathogeny,so that its ability to resist disease and muscle quality are reduced.Gln is a conditionally necessary amino acid,which has not only nutrition,but also important pharmacological action.The thesis sets up the mode of the immunological stress in Trionyx sinensis by using E.coli LPS,and makes an on-the-spot investigation into the protection and mechanism of Gln on internal secretion of the mechanism,ability to resist oxidation,immunologic function, and production performance of the immunological stressed Trionyx sinensis by injecting and adding Gln in the fodder.In order to profoundly probe into immunological stress mechanism,the application of Gln to strengthen the immunological protection lays a theoretical foundation for the healthy cultivation of Trionyx sinensis.In the thesis,seven tests are set for the research on easing of immunological stress mechanism in Trionyx sinensis.Forty-two healthy Trionyx sinensises(339.46±25.54)g whose weight is close to each other are chosen in test one and divided into seven groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.Each Trionyx sinensis was respectively injected into different doses of LPS from abdominal cavity,and heads were cut to get plasma after 5h. COR content in the blood plasma was measured using radioimmunoassay.It is proved that the plasma COR level content in group E is strikingly higher than that in group A(comparison group),B(50μg/kg BW) and C(100μg/kg BW)(P＜0.01),and is much higher than that in group D(250μg/kg BW)(P＜0.05).Thirty healthy Trionyx sinensises (329.66±23.98)g are chosen in test two and divided into five groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.We injected 0.7%NaCl in the comparison group and 500μgLPS/kgBW in each test group respectively,and cut their heads to get plasma at the same time after 5h,1d,4d and 7d.We measured COR,TNF-α,IL-1βand IL-6 content in each sample blood plasma using radio-immunoassay, and IgM,IgG and IgA content in the sample blood plasma using immunoturbidimetery.It is proved that when injected 500μgLPS/kgBW in the abdominal cavity of mature Trionyx sinensis for five hours,the COR,TNF-αand IL-1βcontent is much higher or strikingly higher than that in the comparison group(P_COR＜0.01,P_TNF-α＜0.05,P_IL-1β＜0.01),if injected for 1d,IL-6 content in the blood plasma is strikingly higher than that in the comparison group(P＜0.01),if injected for 4d,the IgM content in the blood plasma is much higher than that in the comparison group(P＜0.05).Fifty-four young Trionyx sinensises(106.46±19.53) g are chosen in test three,and divided into three groups at random,with 3 subgroups and 6 Trionyx sinensis in each subgroup.They were respectively injected into 0.7%NaCl,500μgLPS/kgBW and 1000μgLPS/kgBW.We tested weight gain ratio(WGR) and feed conversion ratio(FCR) of each group after two week's test.It is proved that there is neither obvious difference in WGR and FCR of young Trionyx sinensis in comparison group and group 500μg LPS/kg BW,nor obvious behavior difference in the process of feeding.However,young Trionyx sinensis in group1000μgLPS/kg BW has weakened food intake ability and remarkably decreased WGR(P＜0.01).Furthermore, there is strong reaction,frequent aggression of mutual biting skirt lateral after injection.In conclusion,through COR content change in the blood plasma of mature Trionyx sinensis by injecting different doses of LPS after 5h(test one),change of the COR,TNF-a,IL-1β,IL-6,IgM,IgG and IgA content in th blood plasma at different periods after injection of 500μg LPS/kg BW(test two),and changes in the food intake,growth and behavioristics of young Trionyx sinensis after injection of different doses of LPS(test three),the injection of 500μgLPS/kg BW into the abdominal cavity could cause immunological stress reaction in mature Trionyx sinensis,while injection of 1000μg LPS/kg BW into the abdominal cavity could inhibit its growth.in young Trionyx sinensises and COR content in the blood plasma could be regarded as one significant index to supervise the immunological stress reaction of Trionyx sinensis.Sixty Trionyx sinensises(317.86±22.57) g are chosen in test four and divided into three groups at random,with 4 subgroups and 3 Trionyx sinensis in each subgroup in group A,with 8 subgroups and 3 Trionyx sinensis in each subgroup in group B and C.group A,B and C were respectively injected 0.7%NaCl,500μgLPS/Kg BW,500μg LPS+100mgGln /Kg BW into abdonamal cavity to inspect the content variation of COR,TNF-α,IL-1β,IL-6,GSH,GSH-PX,MDA in blood plasma and protein,GSH,GSH-PX,MDA in lever at different times.The result indicates that after injection of LPS,the content of COR,TNF-α,IL-1β,IL-6,MDA in blood plasma and PRO,MDA in the liver organization strikingly rise at the corresponding period(P＜0.01),while the GSH content in blood plasma remarkably lower(P＜0.05).The supplement of Gin not only eases the rising trend of COR,TNF-α,IL-1βand IL-6,the decrease of GSH in the blood plasma,but also prominently lowers the MDA level of liver.Gln could protect the immunological stressed Trionyx sinensis by restraining the secretion of concerned cell factors such as TNF-α,IL-1β,IL-6,glucocorticoid(COR) and improving the oxidatin resistance ability. Seventy-two Trionyx sinensises(332.35±13.75) g are chosen in test five and divided into four groups at random,with 6 subgroups and 3 Trionyx sinensis in each subgroup.Group A and B are fed with basal feed,group C and D are fed with basal feed plus 2%Gln.After being fed for one week,group B and C are injected into 500ugLPS/kgBW,group A and D are injected into 0.7%NaCl.4d,11d and 25d After injection the content of CD₄⁺/CD₈⁺ in peripheral blood,transformation ratio of T lymph,IgM,IgG,IgA in the blood plasma, muramidase and S-IgA content in intestinal canal were examined.The result proves that the immunological stress caused by LPS strikingly improves the IgM level(P＜0.01) in the blood of Trionyx sinensis.Supplement of Gln not only improves the immunologic function of cells and intestinal canal,but also prohibits the rising of IgM level in the blood plasma conducted by LPS.Thirty-six Trionyx sinensises(318.53±10.26)g are chosen in test six and divided into six groups at random,with 2 subgroups and 3 Trionyx sinensis in each subgroup.We used LPS,Gln and Q to treatt them.After 6h,we got spleen,liver and intestinal canal organization and inspected the expression of HSP70 by immunochemistry.At the same time,we inspected the COR content variety of each group. It is proved that injection of 500ugLPS/kgBW remarkably improves the expression of HSP70 in spleen and liver organization of Trionyx sinensis,while injection of 100mgGln/ kgBW or 8mgQ/kgBW inhibits the expression of HSP70 in intestinal canal,spleen and liver organization.To sum up,Gln could protect the immunological stressed Trionyx sinensis by regulating the immunological function,inhibiting the secretion of inflammatory cell factor and improving the oxidation resistance ability,while the protection might have nothing to do with the expression of HSP70.Seventy-two Trionyx sinensises(101.92±17.53) g are chosen in test seven and divided into four groups at random,with 6 subgroups and 3 Trionyx sinensis in each subgroup.Group A and B are fed with basal feed,and group C and D are fed with basal feed plus 2%Gln.On the eighth day,group B and C are injected into 1000ugLPS/kg BW and 0.7%NaCl is injected into group A and D.On 51th day,they are under the same treatment,on 95th day,we inspected the weight gain ratio,comparison growth ratio, feed conversion ratio,the content of body fat ratio,body skirt ratio,muscle moisture, coarse protein,coarse fat and amino acid,and got small intestine,liver and spleen to observe their structure variation.It is proved that injection of 1000ugLPS/kgBW caused the damage of the villus growth of the intestine,the shrinkage and degeneration of liver cells,which strikingly inhibits the growth of the young Trionyx sinensis and lowers the quality.Supplement of 2%Gln could improve the growth of intestine,remarkably enhance the growth,restrain the damage of immunological stress to the liver of Trionyx sinensis,and improve the production performance of Trionyx sinensis reduceded by immunological stress.