| This study is divided into three parts. In the first part Apostichopus japonicus is used as a test object. Pharmacokinetics of rhein was studied on coelomic fluid, respiratory trees, muscle and integument of Apostichopus japonicus after single dose injection, providing a theoretical guidance for the production of practical application. The second part is about pharmacokinetics of rhein in Penaeus vannamei after a single intramuscular injection. The concentrations of rhein in muscle, hemolymph, hepatopancreas and gill were detected, which can provide a basis for rhubarb and its preparation in the treatment of shrimp gill-rot disease. The third part is studied the effects of rhubarb on non-specific immune in Apostichopus japonicus, guiding the use of rhubarb in the aspect of immunization on Apostichopus japonicus. The main results are as follows:Part one: Studies were conducted on the pharmacokinetics of rhein in Apostichopus japonicus. Coelomic fluid, respiratory trees, muscle and integument were sampled after coelomic injecting with a single dose of rhein 5.33mg/kg. The contents of rhein were determined by high performance liquid chromatography. The results showed that the peak time (Tmax), elimination half-time (T1/2β) and the area under the concentration-time curve (AUC) in Coelomic fluid, respiratory trees, muscle and integument were as follows: Tmax 0.26h, 0.67h, 0.54h and 0.88h ; T1/2β6.24h, 26.1h, 71.48h and 8.93h; AUC 69.29mg/L?h, 105.6mg/L?h, 132.38mg/L?h and 20.99mg/L?h. The concentration-time course of rhein in coelomic fluid, respiratory trees and muscle could be described by a two-compartment model, except for the course in integument being described by a one-compartment model. These results are suggesting that rhein can reach the peak time with a high speed absorption in Apostichopus japonicus after injection, while it is slowly metabolized with a long elimination half-time. Rhein is mainly excreted from respiratory trees.Part two: Pharmacokinetics of rhein in Penaeus vannamei was studied. The concentrations of rhein in hemolymph, gill, muscle and hepatopancreas were determined by high performance liquid chromatography after intramuscular with a single dose of rhein 5mg/kg. The results showed that the peak time (Tmax), maximum concentration (Cmax), apparent volume of distribution (V), elimination half-time (T1/2β) and the area under the concentration-time curve (AUC) in hemolymph, gill, muscle and hepatopancreas were as follows: Tmax 0.17h,0.23h,0.44h and 2.73h; Cmax 1.81mg/L,2.06mg/L,0.44mg/L and 2.31mg/L; V 2.62L/kg,2.33L/kg,10.35L/kg and 0.96L/kg; T1/2β2.13h,3.72h,3.12h and 2.36h; AUC 5.85mg/L?h,11.54mg/L?h,2.18mg/L?h and 17.56mg/L?h. The concentration-time course of rhein in the four tissues could be described by a one-compartment model. These results are suggesting that rhein can reach the peak time with a high speed absorption in Penaeus vannamei after injection, and it is quickly metabolized with a short elimination half-time. Rhein mainly exists in the gill, which provides a theoretical basis on the Rhubarb treatment gill-rot disease in aquaculture production of Penaeus vannamei.The activities of lysozyme (LSZ), superoxide dismutase (SOD), alkaline phosphatase (AKP), and acid phosphatase (ACP) were monitored in the coelomic fluid of Apostichopus japonicus at different days after feeding Rhubarb diet. The contents of rhubarb in diet were 2%,5% and 10%(by mass), besides a control group was set up. The time of feeding Rhubarb was 28 days, after that blank diet was fed. The results showed that the LSZ activity of 10% dose group has doubled the activity of the control group on the seventh day, while 2% and 5% dose group relative to the activity of the control group increased by 92.86% and 96.43% on the twenty-one day; SOD activity of all of the three dosage groups were highest in the seventh day, respectively increased 4.36%, 8.87% and 9.28% than that of the control group, but all of them were lower than the blank Group on the twenty-eighth days with a significant (P <0.05) difference; AKP activities of 2% and 5% dose group were lower than that of the control group, while the AKP activity of the 10% dose group reached the highest value on the twenty-day, which was 6.99 times than that of the blank one, displaying with a significant difference; relative to the control group, the maximum activity of ACP in each group were 3.55 times, 3.79 times and 8.61 times. The results show that rhubarb can increase non-specific immunity of Apostichopus japonicus. We recommend feeding with 2% and 5% of rhubarb for three weeks, containing 10% rhubarb diet two weeks can improve non-specific immunity of Apostichopus japonicus. Stop feeding about two weeks can restore to the normal level of immune parameters. |
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