Part Biological Characteristics Of Microsporum Gypseum From Giant Panda,and The Extraction And Identification Of Extracellular Protease

Objective:To explore the biological characteristics of Microsporum gypseum from giant panda,we study hair perforation test,drug susceptibility test and cultivated the fungus in different culture conditions.Did some extraction,identification and pathogenicity studies to the extracellular protease of Microsporum gypseum from giant panda,to provide a basic information for a further research of the pathogenic mechanism,the diagnosis and treatment to Microsporum gypseum from giant panda.Method:Using different conditions(medium,carbon,nitrogen,temperature,PH)to culture Microsporum gypseum from giant panda,observing the growth morphology and daily growth speed(measured by crossover method).Hair perforation test was carried out to observe the damage degree of hair at different time points.Antimicrobial Susceptibility Test of 96-well Micro-dilution was used to determinate the minimal inhibitory concentrations of 5 drugs.The extracellular protease produced by hair-induced liquid fermentation could be identified by Liquid chromatography-mass spectrometry,the data were analyzed by GO functions enrichment,KEGG Pathway and interaction(PPI)bioinformatics analysis.Result:1.The optimum culture conditions of Microsporum gypseum from giant panda were 25?,pH 10,M-SDA,glucose and peptone,respectively.2.Hair perforation test was cultured 7d,the mycelium was dense,the spores continually grew and sectioned.Becides,the hair damage was serious,some even perforated.3.The MIC value of Fluconazole was 32?g/mL,the Ketoconazole,Ciclopirox,Itraconazole and Griseofulvinwere 1,1,0.5,and 2?g/mL,respectively.The order of sensitive drugs of Microsporum gypseum from giant panda were:Itraconazole>Ketoconazole=Ciclopirox>Griseofulvin>Fluconazole.4.We extracted a number of extracellular proteases which associated with dermatophytes,including ? sulfite reductase,serine proteases,ammonium peptidase and dipeptide enzymes.These proteases could cause skin lesions in mice,including the epidermis cutin and prickle cell flaked,hair follicles and sweat glands in cuticle significantly reduced or disappeared and the inflammatory cells infiltrated extensively.Conclusion:1.The Microsporum gypseum from giant panda could grow in a wide range of culture conditions;Cultivated the Microsporum gypseum from giant panda with hair could cause hair perforation after 7d;The order of sensitive drugs were:Itraconazole>Ketoconazole= Ciclopirox>Griseofulvin>Fluconazole.2.This fungi can secre a number of secretory proteases associated with dermatophytes,including ? sulfite reductase,serine proteases,ammonium peptidase and dipeptide enzymes.It confirmed that extracellular protease of Microsporum gypseum from giant panda could damage the stratum corneum in mice and cause inflammation.