Study On The Quality Standards And The Pharmacology And Toxicology Of The Chinese Traditional Veterinary Compound Preparation "Lian Teng"

This paper was carried out to investigate the quality standards,antibacterial activity,sub-chronic toxicity tests and preventive effects of the Compound preparation "Lianteng" for artificial infection with Salmonella pullorum disease,in order to provide a theoretical basis for development and clinical application of the preparation.Methods:(1)The compound preparation "Lianteng" was composed of Chinese herbal medicine Forsythia suspensa(Thunb.)Vahl and Millettia dielsiana and so on.The effective components of this compound preparation were extracted by using water decoction and ethanol extraction method;Forsythia and formononetin in the extract of compound preparation "Lianteng"were identified by TLC and its content were determined by HPLC,so as to establish the quality standards of the compound preparation "Lianteng".(2)Susceptibility paper method was used to determine antibacterial effect in vitro,and 2-fold tube dilution method was used to determine the minimum inhibitory concentration of the compound preparation "Lianteng" with chicken Escherichia coli,Staphylococcus aureus,respectively.(3)The sub-chronic toxicity of"Lianteng",was determined by giving the preparation with different doses of 2.5,5.0,or 10.0 g·kg-1·bw-1 to the Kunming mice.The organ index,blood ALT,AST,Cr,the white blood cell and leukocyte differential count were examined in the animals.The histological changes in liver,kidney were observed.(4)The"Lianteng" at doses of 1.0,2.0 or 4.0 g·kg-1 was administred in the animals to investigate preventive effect of the preparation for artificially infected pullorum disease in chicken,by researching clinical symptoms,incidence and blood viable count and so on.Results:(1)With the identification of the thin layer,the chromatographic separation was fine and the spots were clear.Within 8?100 ?g·mL-1 of the sample mass concentration of forsythia glycosides,the linear relation was fine(R2=0.9995),the average recovery was 96.40%(n=6);Within 6?150 ?g·mL-1 of the sample mass concentration of formononetin,the linear relation was fine(R2=0.9998),the average recovery was 95.13%(n=6);From the 0.1 g·mL-1 extract of compound preparations "Lianteng",the content of the forsythia glycosides and the formononetin were 47.34 ?g·mL-1 and 35.30 ?g·mL-1.(2)Antibacterial test results showed that the inhibition zone diameter of the concentration of 1 g·mL-1 of compound preparation "Lianteng" extract for chicken Escherichia coli and Staphylococcus aureus were 10.58±1.62 mm and 16.20± 0.42 mm,were moderately sensitive and highly sensitive;the inhibition zone diameter of the concentration of 100 mg·mL-1 of compound preparation"Lianteng" extract for Staphylococcus aureus was 12.35 ± 1.15 mm,was moderately sensitive.In addition,the MIC of compound preparation "Lianteng"extract for chicken Escherichia coli and Staphylococcus aureus were 3.906 mg·mL-1 and 31.250 mg·mL-1.(3)The results of sub-chronic toxicity test showed that the "Lianteng" reduces no significant effect on the weight gain and the feed efficiency of the mice,when compared with the control group(P>0.05);The liver indices of the animals at high-dose of "Lianteng" were significantly increased(P<0.01)and the spleen indices were significantly increased(P<0.05);The kidney indices had not change(P>0.05);The various organs indices of medium-dose group and low-dose group were not significantly changed(P>0.05);The blood AST activity and ALT activity in high,medium and low dose groups were increased,as well as the blood AST activity of the medium-dose group was significantly different(P<0.05),the other groups had no significant difference(P>0.05);There was no change in creatinine level in blood of animals treated with "Lianteng"(P>0.05);There was no change in WBC in animals treated with "Lianteng"(P>0.05),but increased proportion of lymphocytes in the blood of mice was found.There was no significant histological change in liver and kidney.(4)The compound preparations"Lianteng" decreased the mortality of chickens artificially infected with Salmonella,the results showed that the compound preparations "Lianteng" had a prevention function for the pullorosis.Conclusions:(1)In this study the HPLC mass spectrometry method of compound preparation "Lianteng" was established,which could be used in examing quality standards of veterinary medicine compound preparation"Lianteng";(2)The antibacterial effect of compound preparations "Lianteng"extract for Escherichia coli and Staphylococcus aureus were conformed.(3)The results of sub-chronic toxicity test showed that the compound preparation"Lianteng" actual non-toxic,Security was fine;(4)Compound preparations"Lianteng" had a prevention effect against the pullorosis.