| Duck plague which is caused by duck plague virus (DPV) is an acute contagious disease of ducks and goslings,No matter what age group or variety can infect this virus. And the shelduck, muscovy ducks and cotton ducks are the highest infectious. In the natural epidemic, adult ducks and laying ducks have serious morbidity and mortality.Duck virus hepatitis (DVH) which is caused by DHV is a highly lethal infectious disease of duckling. The disease is characterized by acuteness, rapid transmission and high mortality. Most of the deaths occur in 3 or 4 days. Both of the morbidity and mortality are very high on ducklings, even 95% on 1-week-old duck.Preventions are through by immune breeder ducks and ducklings. There also have many active ingredients that traditional Chinese medicine have effect on virus in cells or embryo reported in the literature.In the same time, through recent years'study, the complex traditional Chinese medicine "Qin kangsan" had preventative and therapeutic effect on a number of bacterial and viral disease of birds. Improving immunity was also one effect of "Qin kangsan". So, we proposed the subject that effect research of "Qin kangsan" on duck plague and duck hepatitis virus.In the present study, the anti-DPV and anti-DHV effects of "Qin kangsan" decoction were investigated on two levels:duck embryo, duck embryo fibroblasts. Through the test, we looked forward to accumulating data and providing support for in-depth study of traditional Chinese medicine's effect on duck diseases.Test 1 Antiviral effects of "Qin knagsan" against DPV and DHV in duck embryo. The 48 duck embryo were assigned into 6 groups,8 per group. 100mg or 25mg decoction was inoculated into embryo after being mixed with virus(DHV or DPV) solution of 100ELD50 1h, which called group of Chinese anti-virus. And as for preventing grop,100 mg or 25mg decoction was drew in duck embryo,24h later, virus(DHV or DPV) solution of 100ELD50. Positive control group was only 0.2m virus(DHV or DPV) solution of 100ELD50.Just 0.2ml PBS of negative control group. The protection ratio reached to 100%.87.5% of high dose prevention group (100mg "Qin kangsan") to duck embryo inoculated DHV,DPV. And low dose prevention group (25mg "Qin kangsan") reached to 100%,87.5% The protection ratio reached to 50%,12.5% of high dose of anti-virus group, and 25%,12.50% of low dose of anti-virus group. The result indicated that "Qin knagsan" had prominent killing and prohibiting infection effect on DPV and DHV in 10-day-old duck embryo.Test 2 Study on proliferation effect of "Qin kangsan" to DEF. The decoction diluted to 0,2-4,2-5,2-6,2-7,2-8g/ml was drew in DEF,100μl per plate,4 plates per concentrations.One 96 well plate was took after 12h,24h,36h,48h,60h for getting the OD by method of MTT. The result indicated, the maximum safe concentrations of "Qin knagsan" for DEF was 2-4g/ml. In the concentrations of 2-4-2-8g/ml significantly promoted DEF proliferation. And the 10-5g/ml was the best one.Test 3 Research on the protection effects of "Qin kangsan" to DEF infected by DHV or DPV artificially. We took three ways:①"Qin kangsan"(2-4-2-8g/ml) was mixed with 100TCID50 virus(DHV or DPV) at the same volume.37℃,4h later,200μl mixed liquor was inoculated in DEF;②100μl virus first,4h later, 100μl "Qin kangsan" of every concentration gradient;③The method was opposite with②. The result indicated: every concentration gradient of "Qin kangsan" had effect to remove restraint of DHV,DPV to DEF. "Qin kangsan" of 2-6-2-4g/ml had the best effect to DHV,③was the best way.Cmpared with negative control, difference is extremely remarkable(P<0.01) "Qin kangsan" of 2-5-2-4g/ml had the best effect to DPV, compared with negative control, difference is extremely remarkable(P<0.01),③was the best way. |
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