| The intension of this study is to find out the leading pathogens causing diary cows subclinical andclinical mastits and effective antibiotic treatment to certain pathogens. Several major pathogenicmicroorganism that cause mastitis of diary cows were select, and the antibiosis effects of Chinesemedicinal herbs were tested through the methods of two fold dilution and stiletto. Theanti-inflammatory effects of Chinese medicinal herbs were tested by swelling of mice pinna by xylene,sack of rat by carboxymethyl cellulose and granuloma with cotton ball in mice. The toxic effects ofcompounds of Chinese medicinal herbs were studied through acute toxicity test. The best Chinesemedicinal herb compound was determined on the basises of antibiosis expriment, anti-inflammatoryexpriment and toxicity experiment. The technologied process and the stability of Chinese medicinalherbs ointment were studied, on the basis of Chinese medicinal pharmacy. Permeation quantity in vitroof main composition-caffeic acid in Chinese medicinal herbs ointment were determined by HPLCmethod. The best enhancing penetration concentration of zzone in Chinese medicinal herbs ointmentwere defined. The safety action of Chinese medicinal herbs ointment was studied.The result of epidemiological survey indicated that in a certain herd (105 cows), the cow infectionrate of clinical mastitis is 13.33%;that of subclinical mastitis is 54.29%. The GVFBX quarter infectionrate of clinical mastitis 5.29% and that of subclinical mastitis is 27.06%.49 bacterial strains were isolated from 10 milk samples of subclinical mastitis, includingstaphylococci (16 of 49,32.65%), streptococci (19 of 49,38.78%) and Gram-negative bacilli (14 of49,28.57%). 258 bacterial strains were isolated from 10 milk samples of clinical mastits, includingstaphylococci (65 of 258,25.19%), streptococci (164 of 258,63.57%) and Gram-negative bacilli (28 of258, 11.24%).The result of two fold dilution indicated that the antibacterial activity of formula 3, formula 4,Raidix saposhnikoviae, Fructus chebulae, Radix Scutellariae on 5 pathogenic microorganisms muchbetter, and their MIC reached 4～63g·l-1, MBC reached 8～63g·l-1.The antibacterial activity of RadixIsatidis, Fructus arctii, Radix glycytthizae, Flos Lonicerae, Fructus Forsythiae, Herba Taraxaci on 5pathogenic microorganisms less better,their MIC reached 31～125g·l-1, MBC reached 31～125g·l-1.The result of stiletto indicated that the antibacterial activity of Fructus Chebulae, RadixScutellariae, Fructus Forsythiae, Radix Isatidis, Raidix Saposhnikoviae, Squama Manitis, SemenVaccariae, Radix Glycytthizae, Fructus Arctii, Rhizoma Chuanxiong, Pericarpium Citri, ReticulataeViride, Radix Salviae Miltiorrhiae and Medulla Tetrapanacis were moderate sensitive or extremelysensitive to 5 pathogenic microorganisms;formula 3 and formula 4 were moderate sensitive orextremely sensitive to the pathogenic microorganisms except for Escherichia coli. So the integrativeantibacterial activity of formula 3, formula 4, Fructus Chebulae, Herba Taraxaci, Radix Scutellariae,Raidix Saposhnikoviae, Semen Vaccariae, Rhizoma Chuanxiong, Radix Salviae Miltiorrhiae and RadixIsatidis much better. Formula 3 and formula 4 were chose to do the next experiment.The result of anti-inflammatory experiment indicated that the inhibition rates of granuloma withcotton ball in mice of formula 3 from high concentration to low concentration were 27.53%, 20.32%and 13.12%;that of formula 4 were 29.34%, 21.43%and 12.58%. The inhibition rates of swelling ofmice pinna by xylene of formula 3 were 29.34%, 1.43%and 12.58%;and that of formula 4 were 16.80%, 14.48%and 7.41%. The inhibition to leukoplania from sack of rat by carboxymethyl cellulose offormula 3 and formula 4 much better compared to the control group. So the anti-inflammatory effect offormula 3 and formula 4 much better to various state of active chronic inflammation, the result showedthe dose-effect relationship, and the anti-inflammatory effect of formula 3 much better than formula 4.The result of acute toxicity test indicated that LD50 of formula 3 and formula 4 were6523.79mg · kg-1 and 22861.25mg · kg-1, and its 95% confidence interval were 5513.91 ～7718.63mg·kg-1 and 20641.44～25319.78mg·kg-1. So the formula 3 and formula 4 were practicalnon-toxicity substance, and the formula 4 was safer than formula 4.The formula 4 was determined to further studied on the bases of antibiosis effects experiment,anti-inflammatory experiment and acute toxicity test.The Chinese medicinal herbs ointment didn't appear any phenomena of breaking emulsion,mildewy and the change of color, lustur, homogeneity by cold –resistant, heat-resistant, centrifugal andaccelerating experiment;The amounts of caffeic acid through the skin mice by HPLC method, theresults indicated that the amounts of caffeic acid was highest when the concentraction of azone inChinese medicinal herbs ointment was 4.33%. The permeation amounts of caffeic acid with 6% azonein the Chinese medicinal herbs ointment (0.6317μg) were lower than that with 4% azone (0.8041μg);The result of irritative and anaphylaxis experiment showed that Chinese medicinal herbs ointment werenon-irritation or weaker anaphylaxis substance. When it was applicated externally, the score of skinirritative action were smaller than 0.5, and the anaphylaxis rate of Chinese medcinal herbs ointmentwas 0%;The results of Clinical Pharmacodynamics experiment showed that the ointment had goodtherapeutic action on clinical mastitis of diary cattle.Above all, the studies showed that traditional Chinese medicines ointment was a good skinpermeating preparation, which was non-toxic, non-irritating, non-anaphylaxis,and had good curativeeffects on clinical mastitis of dairy cow.
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