Cultivation, Drug Screening Of E.wenyoni In Vitro And Therapeutic Test Of Sick Cows

Different proportion of NBCS,glylose and NaHCO₃ were added into the mediums of RPMI-1640,MDEM,M-199 and HL,those were used to compwere the effects of E.wenyoni in vitro in order to definite the optimal medium and the optimal proportion of NBCS, glylose and NaHCO₃ .The serial subcultivation of E.wenyoni in vitro and the expriment of that erythrocytes of human,rabbit,goat and swine were infected by E.wenyoni were undertaked which according to the results of this study.In order to filter the best medicine that can kill E.wenyoni in vitro ,the kill experiment in vitro of imidocarb,primachin phosphate, oxytetracycline hydrochloride, tetracyline and so on,were undertakend by the conditions which were definited according to the results of culture in vitro.Through comprensive evaluation,two medicines were chosen to cure the Eperythrozoonosis.The choice drug which was used to cure Hydrochlorideure the Eperythrozoonosis was definited by the conventioal observation of clinic,the microscopic exmination of the blood smears,index of the physiology and biochemistry of the blood.The results suggested that under the conditions of 37°C,5%CO₂ and the medium were renoreatured every 12 hours,the erythrocytes had a higher in fection rate and there were more E.wenyoni on their surfaces,when they were cultured in the medium of RPMI-1640 which was added in 30% NBCS,2% glylose and 0.20%NaHCO₃ or in the medium of HL which was added in 20% NBCS and 1%HEPES.The statwastical results were no significant between these.But the RPMI-1640 was the optimal medium because the cose of the HL and HEPES were higher than RPMI-1640.When the infection rate was more than 80%,the erythrocytes of these apertures were collected and 2μl erythrocytes were remained in every aperture.Every aperture was supplied with 18 μl normal cow erythrocytes and some complete medium in order to make the infection rate of the erythrocytes retain 7～9%,After that these erythrocytes were cultured under the conditions of primary culture in vitro and they could serial passage more than 20.The erythrocytes of human, rabbit,goat and swine can infect E.wenyoni when they were cultured in vitro,Thwas indicated that the E.wenyoni had not severe and specitied host,and the infected rate of the rabbit RBC was higher than others.The experiment of kill in vitro indicated that the kill effect of imidocarb and primachin phosphate were the best ,and they had less influence on erythrocytes than dipterex, ethacridine and etc.Therefore, imidocarb or primachin phosphate should be the choice drug to cure Eperythrozoonswas, dipterex and ethacridine were the second choice.The result of the therapeutic test indicated that the cure effect of primachin phosphate was better than imidocarb,and its secondary effect was less ,thus primachin phosphate was the choice medicine through the obervation of clinical symptom,the microscopric examination of the blood smears and the determintion of physiology and biochemwastry of the blood.