Research Of Microbial Diversity And Isolation Of Deodor Microorganism Of Earthworm Cast

With the development of intensive Animal Husbandry,odor emitted from Animal Husbandry not only affected the health and production performance of livestock,but also threated the health of people,earthworm cast,which has high factor of porosity and specific surface areais,is deemed to be a effective adsorbing agent of odor.Earthworm cast can deordor by decomposed the organic matter.In 1980's, earthworm cast was used to dealt with the domestic waste in Germany and Japan. Untill now,research in this area is still relatively small In domestic.In this paper.Research on Biological deodorant of earthworm cast was carried out as follows:1.With cow dung as stock blend,the mixture of upper,middle and lower classes of earthworm cast were collected respectively in keeping to the first 10 d,20d,30d. Among DNA exrtacted mhetods,total DNA was successfully extracted from the feces of the earthworm by using acetone to remove feces organic matter and Proetinase K,SDS,phenol to lyses the cells.Integrity and purity of DNA was confirmed by spectrophotometer and electrophoresis.The DNA,which in length was 23kb and the absorption of which is between 1.6 and 1.8,was suitable to be analysed by PCR-SSCP.2.With PCR-SSCP methods,microbial diversity of the earthworm cast of different stages were analyzed.Itis found that the diversity of bacteria line up before 20 d,but,with the lapse of time and the base material was slowly consumed,the diversity of bacteria presented in the slow upward trend and the diversity of fungi presented in a declining trend.The diversity of the bottom bacterial microbial was higher than that of any other microbial at the level of index,and exceeded 0.8.3.Through the enrichment and domesticated of microbial,20 deodorant microbes was isolated,including seven bacteriaswhich could decompose H₂S,four fungi which could decompose H₂S,six bacterias which could decompose NH₃,three fungi which could decompose NH₃.With further screening experiment,11 stains deodorant microbes was isolated.The bacteria accounted for 55 percent of total isolated microbes from the result.4.With a non-antagonistic B6 and B1-6 as tested strain,orthogonal variance analysis was carried out to Screen the compost formula of deodorant assay.The proportion of material of compost:80 percent of pig manure,straw 20 percent,the ratio of microbes and material 1:1/1000.5.According to the result of orthogonal experiment,non-antagonistic three strains micorbes of decomposing H₂S and NH₃ were mixed to deordor of compost. Experimental results showed that:18d in culture,H₂S emissions the average emissions is 213.13 mg/kg,.the average emissions decreased 46.26%,compared with the CK,respectively.NH₃ emissions the average emissions was 146.74mg/kg.the average emissions decreased 46.31%,compared with the CK,respectively.Compared the deodorant effect between three strains and single strain in same time,H₂S emissions and the grade of odor of three strains was superior to that of all a single strain.6.The microbes was confirmed with molecular biology technology.The result showed that B1,B5,B1-2,B1-4 belonged to Bacillus and B3,B2 belonged to Alcaligenaceae and F3 belonged to penicillium and B1-6 did not belong to any knowed genus.Bacillus accounted for 57.2%of confirmed microbes and belonged to predominant microbes to deodor.