Bacteriostatic Test In Vitro And The Experimental Animal Safety Evaluation Of Gamithromycin

Objective: Macrolide antibiotics play an important role in the treatment of respiratory tract infection.The animal medicine mainly through the 50 S subunit of the bacterial ribosome binding and inhibition of protein synthesis.The clinical use of drugs in the treatment of respiratory tract infection have different limitations,with the new resistant strains emerge,imminent development of new macrolide antibiotics.France Merial(MERIAL)developed by Gamithromycin is a special kanamycin antibiotic animal,the drug in 20072010 and 2011 respectively by the European Medicines Agency,Canadian health care products and food administration and the U.S.Food and Drug Administration approved for the prevention of mannheimiahaemolytica,kill bacteria and sleep histophilus caused by cattle respiratory disease of PAP.Due to Gamithromycin with the rapid absorption,wide distribution,high concentration of lung tissue in eliminating slow,sustained effect,high security,try Jamie azithromycin injection in Tianjin or the challenge of Bio Science and technology research and development,the test of Tianjin Zhongsheng R & D pilot Gamithromycin activity of azithromycin injection laboratory evaluation,toxicity,safety evaluation and other aspects,to provide basic data for drug development and reporting number.Methods: 1 To collect clinical lung symptoms of respiratory tract disease in cattle,tracheal effusion and subcutaneous gelatinous liquid infiltration,under sterile conditions,inoculated in BHI broth,after purification,bacterial morphology observation,biochemical identification and culture characteristics,and analysis of bacterial 16 S rRNA sequences to determine the treatment of bacteria species,16 S rRNA by the method of PCR amplification,sequencing analysis;to determine the species of bacteria using bacterial species specific gene Kmt-1.2,The CLSI's method,test of azithromycin injection in Garmisch has been isolated from bovine mycoplasma pneumonia in the determination of trace two times dilution method,minimal inhibitory concentration of Pasteurella multocida and hemolytic manesque coli(MIC).3,Using the modified Karber's method of 66 clean grade Kunming healthy mice median lethal dose(LD50)were determined in mice;continuous subcutaneous administration of 28 d,to observe the long term toxicity on mice,in Gamithromycin injection into high and low,three doses(219?109.4?10.94mg/kg)the solvent group on organ coefficient and histopathology were observed.4,In accordance with the "chemical irritation,allergic and hemolytic technical research guidelines >(2005)of rabbit skin irritation test in rabbits,the injection site irritation test,hemolysis test,active systemic anaphylaxis test and guinea pig passive allergy test.Results: 1,Lung disease of cattle disease,isolated 3 strains of dew like,the trachea and subcutaneous effusion jelly like liquid infiltration without hemolysis,gray white colonies,by Gram staining microscopic examination,can be observed in gram negative coccobacilli,through biochemical test instrument Merieux,and related test a(2 VITEK COMPACT)for biochemical identification of the strains of Pasteurella multocida 16 S rRNA sequence homology of 99% result for kill Bacillus.16 S rRNA sequence analysis and NCBI of PAP has announced.The separation of the PAP Bacillus were injected into mice,all mice died.The results showed that the isolated from the cow in 3 strains of Pasteurella multocida.2,In Gamithromycin of control bacteria and Mycoplasma antimicrobial effect,minimal inhibitory concentrations were 0.5 g/mL,and 1 g/mL,in accordance with the quality control range < animal bacterial antimicrobial susceptibility test paper method dilution method on the implementation of Standards >(0.5-4g/mL),and CLSI < broth dilution method and agar dilution method for determination of Mycoplasma pneumoniae MIC(g/m L)to explain the discount range on the quality control guidelines >(0.5-1 g/m L).3,azithromycin injection LD50 1094.2mg/kg rice,95% confidence limit of 1000 mg/kg-1196.7mg/kg28d;continuous subcutaneous drug for chronic toxicity showed examination and control group animal organ coefficient and pathological differences were not statistically significant(P>0.05).4,the results of in vitro hemolysis test were negative,and the stimulation test was negativeConclusion: We have successfully isolated from bovine Pasteurella multocida disease in vivo.Test of azithromycin injection in Garmisch causing bovine respiratory disease mannheimiahaemolytica has high antibacterial activity and Pasteurella multocida and mycoplasma.According to the acute toxicity of chemical LD50 dose of azithromycin injection grade show Garmisch belong to the low toxicity of drugs.Combined with the stimulation test,hemolysis test,systemic anaphylaxis test results show that,in Gamithromycin injection will not have adverse effects on the target animal.