Distirbution Of Microbe And Greenhouse Gas In Different Depth Of Bio-Bed

There are numerous microbes in the litter of bio-bed, these microbes were the most important factors. This study use PCR-DGGE, Real-Time PCR technique and infrared acoustic spectrum monitor, to analyze flora similarity, the number of Escherichia coli, lactobacillus, methanogens, and concentration distribution of CO2,CO,N2O,NH3,CH4, H2O in different depth of bio-bed for the further study of community ecology on bio-bed.Due to the complex compositions of the litter in Bio-bed, there are many impurities in DNA extract. Under these circumstances, the universally applicable and reproducible method to extract DNA from different organisms is not desirable. Thus, the aim of this study was to modify the conventional method in order to extract high quality complex genomic DNA from different bacteria in Bio-bed. Our results demonstrated that modified Genoise Bacteria DNA Ectraction kit can remove PCR inhibiting factor effectively,and the optimal concentration of thiamine hydrochloride was0.5M.Through the test of bacterial universal primer968f/1401r,bacterial16S rDNA sequences,which were samples from the litter in bio-bed of Taicang pig breeding farm, were amplified by PCR,and using the DGGE fingerprint technology to analyse the products of the PCR.It was found that the number of bacterial bands in a higher level (0-15cm) of litter in bio-bed was similar and the fluctuation range was from24to27.The number of bacterial bands in the depth of20cm was the lowest,while the number in the depth of30cm reached the maximum,The similarity between the depth of10cm and other depths of litter in bio-bed was the lowest,only51.2%,followed was the surface layer of padding,and the similarity of residual levels was56.8%;the similarity of the surface and the depth of5cm was the closest, reaching95.6%.By Real-Time PCR technology, this paper made a quantitative analysis of E. coli, Lactobacillus and Methanogens of litter in different depths on bio-bed.The study found that the largest number of E. coli was on the surface of bio-bed,which was up to9.68×105cfu-g-1. With the depth increasing,the number of E.coli reduced, the numbr at40cm was least. Thus, we can found that bio-bed could effectively control the number of E. coli, so, if we want to extend feces-decomposition area, reduce the growth of escherichia coli, we should try to extend function area and build a approriate oxygen environment in bio-bed. Lactobacillus were founded in different depth of bio-bed, and the number of lactobacillus on the surface of bio-bed was less, only3.69x103cfu-g"1. With the increase of the depth, the number of lactobacillus increased, at the15cm, the number was6.08×103cfu-g-1which was up to maximum, while, the number was lowest at40cm,onlyl.62×102cfu-g-1.thus, lactobacillus was mainly active in the upper of bio-bed.The number of methanogens are few in different depth of bio-bed, at the range of0～25cm, the number increased with the depth increasing, at30cm, the number declined slightly, at35cm, the number was up to5.08×103cfu-g-1which has on the top. Thus, methanogens was mainly active in the lower area,namely anaerobic zone in bio-bed.In this study, INNOVA1412Infrared Sound Spectrum Monitor was used to monitor six greenhouse gases (CO2, CO, N2O, NH3, CH4, H2O) at different depths of padding material in the fermentation bed. The results showed that with depth gradually increased, CO2, CH4concentration increased, and the increase depended by breeding density, litter’s using time, the gap of plowing time and management, we found that we could control breeding density and management to extend the function distribution of bio-bed.The result also showed that the growth of CO and N2O roughly same with increasing depth,and the concentration of both were obviously impacted by oxygen content,breeding density and management. At the same time,we also found that the emission of NH3was relative to material of litter, management and the contents of H2O in different depth of bio-bed are relatively steady.