| Bovine mastitis is of redness,swelling,heat and pain on mammary glands,which is caused by various pathogenic microorganisms,physical and chemical factors.The disease causes disorders of the microcirculation of the mammary gland,pathological changes of the mammary glands,and the decrease of the immune function.A variety of different types of pathogenic infection of bovine mammary glands are one of the main reasons of bovine mastitis.The disadvantages of antibiotics and other chemicals in the treatment of diseases have been revealed.More and more new types of veterinary drugs have been developed for the reason that Chinese traditional medicines have the advantages of low toxicity or no toxicity,no bacterial resistance,no veterinary drug residue and so on.Qinhong breast perfusion agent is a prerequisite for antibacterial activity test in single herbs.According to the treatment principle of bovine mastitis of traditional Chinese medicine,the amount of drugs were determined by orthogonal test,the optimum extraction conditions were determined by HPLC.Objective: Isolation and identification of the main pathogenic bacteria causing bovine mastitis;To study the bacteriostatic effects,analgesic effects and anti-inflammation effects of Qinhong breast perfusion agent;The acute and subchronic toxicity tests were conducted to investigate the safety of Qinhong breast perfusion agent.Methods: 92 Milk samples from large dairy farm in Inner Mongolia,Chifeng city,were collected.The culture of milk samples,isolation and identification of bacteria were carried out in the laboratory.To determine whether the isolated bacteria were pathogenic bacteria or not through the pathogenicity test on mouse.The bacteriostatic activity,the minimum inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC)were studied by the “Oxford cup method” and “Tube dilution method” in vitro.The analgesic effects of Qinhong breast perfusion agent were studied by “writhing test” and “hot plate test”.The anti-inflammatory test was carried on the swelling of auricle of mouse inducedby the dimethylbenzene.Through the acute toxicity test,the death of the decimal number in the maximum dose test and the the rats’ general living condition,weights,hematological indexes and biochemical indexes,organ coefficients change in subchronic toxicity test to investigate the toxicity of the Qinhong breast perfusion agent.Results: 73 strains of Staphylococcus aureus(43.71%),41 strains of Streptococcus agalactiae(24.55%),22 strains of Streptococcus dysgalactiae(13.17%),31 strains of Escherichia coli(18.56%)were isolated from 92 milk samples.The bacteria have certain pathogenicity.The "Oxford Cup" method showed that the 3 dose groups of Qinhong breast perfusion agent had certain bacteriostatic effect,and the greater the concentration of drugs,the better the antibacterial effect.The MIC of Staphylococcus aureus,Escherichia coli,Streptococcus agalactiae and Streptococcus dysgalactiae were 31.25,62.5,62.5,125.0 mg/mL,MBC were 31.25,125.0,62.5 and 125.0 mg/mL.The results of analgesic test showed that compared with the normal saline group,there was a significant difference in the number of writhing reactions between the high dose group and the medium dose group(P < 0.01),the effect of drug analgesia was positively correlated with the concentration of drugs.In the hot plate method,there was a significant difference between the low dose group and the high dose group at 60 min and 90 min(P < 0.05),and there was no significant difference between the low dose and saline group at 120 min(P > 0.05),while there was a significant difference between the high dose and the medium dose between saline group(0.01 < P < 0.05),it showed that the analgesic effect of the low dose group was short.The anti-inflammatory test showed that the 3 groups had certain swelling inhibition,the difference between the high and medium dose group and the normal saline group was extremely significant difference(P < 0.01),and the swelling inhibition was dose dependent.The acute toxicity test showed that the LD50 of the drug could not be measured.The maximum dose was 80 g/kg,and the maximum tolerated rate was 2000.Subchronic toxicity test results showed that the rats had a normal life,increased weight.The hematological indexes,biochemical indexes and organ coefficients of rats were not affected.Conclusions: Qinhong breast perfusion agent has good antibacterial,analgesic and anti-inflammatory effects.The medicine has no toxicity and side effect,and has high safety. |
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