Practice Instruction of Innovative and Entrepreneurial Training Program for Aquatic Animal Medicine Students

As an important platform for the combination of theory and practice,college students' innovative and entrepreneurial training program has an important impact on improving the comprehensive ability of undergraduates. The major of aquatic animal medicine is a new sea-related major. The continuous development and transformation and upgrading of aquaculture in the new period have put forward newer and higher requirements for the innovative and practical ability and comprehensive quality of aquatic animal medicine graduates. Taking the innovation and entrepreneurship program of college students majoring in aquatic animal medicine as an example,this paper analyzes the significance of the innovation training program for college students,the cultivation of students' scientific research ability,and the problems existing in the application and operation of the program,in order to provide a reference for the innovative education and talent training for students majoring in aquatic animal medicine.

Establishment and Optimization of Two-dimensional Electrophoresis System for Spleen Proteome of Sillago sihama Forsskal

Taking Sillago sihama Forssk?l as the research object,through the optimization of the extraction method,hydration and isoelectric focusing,a two-dimensional electrophoresis system was established for spleen proteome of S. sihama Forssk?l. The results showed that the twpstep hydration and isoelectric focusing method is better than the hydration and isoelectric focusing integrated method. The protein spots of the spleen of S. sihama Forssk?l were detected by two-dimensional electrophoresis mainly at pH 4. 0-7. 0. The establishment of this technical system will lay a foundation for further research on proteomics of S. sihama Forssk?l in the future.

Inhibitory Activity of Prunus mume,Copitis chinensis and Crataegus pinnatifida on Vibrio harveyi and Its Biofilm

[Objective] This study was conducted to determine the in-vitro inhibitory effects of Prunus mume,Coptis chinensis and Crataegus pinnatifida on Vibrio harveyi and its biofilm. [Method]The inhibitory zone diameters of the three Chinese herbal medicines against V. harveyi were determined by agar diffusion method; the minimal inhibitory concentration( MIC) and minimal bactericidal concentration( MBC) values of the three Chinese herbal medicines against V. harveyi were determined by doubling dilution method; and the effects of the three Chinese herbal medicines on the formation of V. harveyi biofilm were determined by methyl thiazolyl tetrazolium( MTT) method. [Result]The three Chinese herbal medicines all inhibited V. harveyi to different degrees. C. chinensis and C. pinnatifida and P. mume exhibited the inhibitory zone diameters of( 17. 62 ± 0. 04),( 20. 16 ± 0. 08) and( 30. 76 ± 0. 26) mm against V. harveyi,respectively. P. mume and C. pinnatifida had strong inhibitory effects on V. harveyi. The MIC and MBC values of P. mume against V. harveyi were 7. 812 5 mg/ml; the MIC and MBC values of C. pinnatifida against V. harveyi were 31. 25 mg/ml; and the MIC and MBC values of C. chinensis against V. harveyi were 62. 5 mg/ml. P. mume had the strongest antibacterial and bactericidal ability. The MIC values of C. pinnatifida,C. chinensis and P. mume against V. harveyi were 7. 81,7. 81 and 1. 96 mg/ml,respectively,i. e.,P. mume exhibited the lowest MIC. [Conclusion] P. mume,C. pinnatifida and C. chinensis all have inhibitory effects on V. harveyi and its biofilm,and P. mume has the strongest bactericidal ability.

Inhibitory Effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio parahaemolyticus and Its Biofilm in Vitro

In order to explore the inhibitory effects of Prunus mume,Crataegus pinnatifida and Coptis chinensis on Vibrio parahaemolyticus and its biofilm in vitro,the agar diffusion method was applied. These three Chinese herbal medicines had different inhibitory effects on V. parahaemolyticus. The inhibition zone of C. pinnatifida to V. parahaemolyticus was( 15. 25 ± 0. 53) mm,and the minimum inhibitory concentration( MIC) and minimum bactericidal concentration( MBC) of C. pinnatifida on V. parahaemolyticus were both 31. 25 mg/m L; the inhibition zone of C. chinensis to V. parahaemolyticus was( 18. 08 ± 0. 10) mm,and the MIC and MBC of C. chinensis on V. parahaemolyticus were both15. 63 mg/m L; the inhibition zone of P. mume to V. parahaemolyticus was( 28. 99 ± 0. 47) mm,and the MIC and MBC of P. mume on V. parahaemolyticus were both 7. 81 mg/m L. The effects of three traditional Chinese medicines on the biofilm formation of V. parahaemolyticus were tested by MTT colorimetric method using methylthiazolyl tetrazolium( MTT). P. mume,C. pinnatifida and C. chinensis have significant inhibitory effects on V. parahaemolyticus biofilm and their MIC are 7. 81 mg/m L,3. 125 mg/m L,and 62. 5 mg/m L,respectively( P < 0. 01).The experimental results are expected to provide certain references for the development of new fishery drugs.

Inhibitory Effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio alginolyticus and Its Biofilm In Vitro

[Objectives]To explore the inhibitory effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio alginolyticus and its biofilm in vitro. [Methods] The agar diffusion method was applied to determine the inhibition zone diameter of three Chinese herbal medicines on V. alginolyticus; double dilution method was used to determine the minimum inhibitory concentration( MIC) and minimum bactericidal concentration( MBC) of three Chinese herbal medicines on V. alginolyticus; methylthiazolyl tetrazolium( MTT) method was applied to evaluate the effects of three Chinese herbal medicines on the biofilm formation of V. alginolyticus. [Results]The three Chinese herbal medicines had different inhibitory effects on V. alginolyticus. The inhibition zones of P. mume,C. chinensis,and C. pinnatifida on V. alginolyticus were( 18. 31 ± 0. 29) mm,( 20. 31 ± 0. 30) mm,and( 30. 24 ± 0. 14) mm respectively,and P. mume had strong bacteriostatic effects on V. alginolyticus. The double dilution method was used to determine the bacteriostatic effects of three Chinese herbal medicines on V. alginolyticus. Results showed that P. mume has the highest antibacterial and bactericidal ability. The MIC and MBC of P. mume,C. pinnatifida,and C. chinensis on V. alginolyticus was 3. 91,31. 25,and 125 mg/m L,respectively. Finally,MTT method was applied to evaluate the effects of C. pinnatifida,C. chinensis,and P. mume on the biofilm formation of V. alginolyticus,and results showed that the MIC on the biofilm formation of V. alginolyticus was 15. 63,7. 81,and 3. 91 mg/m L,respectively; the MIC of P. mume was the lowest. [Conclusions]This experiment proves that P. mume,C. chinensis,and C. pinnatifida have inhibitory effects on V. alginolyticus and its biofilm formation. It is expected to provide references for prevention and control of pathogenic vibrio induced diseases of aquatic animals.

Analysis of Biological Activity and Immunogenicity of Extracellular Products from Streptococcus iniae

[Objectives] This study aimed to analyze the biological activity and immunogenicity of extracellular products from Streptococcus iniae .[Methods] S. iniae was incubated with brain heart infusion agar medium （BHIA ＋ 4% calf serum） for 60 h. The bacterial liquid was rinsed with PBS, centrifuged, and filtered through microporous filtering film to collect extracellular products （ECPs）.[Results] Extracellular proteinase （ECPase） of S. iniae exhibited amylase, protease, lecitinase, gelatinase, lipase activities and hemolytic activity but had no urease activity. EDTA, DTT and PMSF could reduce ECPase activity to 72.4%, 77.6% and 72.4%, respectively. Cu^2＋ , Ca 2＋ , K^＋ and Mg^2＋ exhibited an inhibitory effect on ECPase activity, whereas Fe^3＋ , Co^2＋ and Mn^2＋ could activate ECPase activity. ECPs had good heat stability and exhibited relatively high activities under alkaline conditions. The optimal temperature for ECPs was 55 ℃. Two-dimensional electrophoresis was performed to analyze the main protein of ECPs. The results indicated that there are 12 main bands of ECPs, and the molecular weights mainly ranged between 28-68 kDa. About 120 protein spots were detected, and the molecular weights mainly ranged between 26-95 kDa. The mouse anti- S. iniae was used for Western-blot analysis of ECPs, and the results showed that there were four proteins, with molecular weights of 26, 37, 95, and 97 kDa, respectively. The pathogenicity assay indicated that ECPs of S. iniae were highly pathogenic to tilapia. The mortality rate of tilapia was enhanced as the concentration of ECPs increased.[Conclusions] This study provided a certain theoretical basis for revealing the pathogenic mechanism of Streptococcus iniae .