Detection For Piggery Air Microbial And Research On Contamination And Propagation Problems By 16SrDNA High-throughput Sequencing

Abstract Air microorganism is one of the important factors of intensive pig farms environment pollution,The pathogenic microorganisms in the air can affect the health of animals, workers and nearby residents, and airborne microorganisms may be derived from fecal bacteria atomization. Currently, researches on pig airborne microbes mainly concentrated on a few specific microbes or pathogens. The study on the microbial diversity and abundance of the piggery air or air pollution around the piggery and comparative studying on microbes of air and fecal are less.Therefore, The experiment was carried out on Qionglai, Sichuan Jinli pig farm from July to August in 2013. Air samples were collected on the spots at seven different distance from pig houses with a liquid impinge sampler. The total DNA of air samples and fecal samples were extracted, then the DNA samples were sent to Novogene Bioinformatics Institute, Beijing to sequence.The QIIME Software was used to analyze the microorganism abundance and diversity, a and (3 diversity analysis shows that the difference of abundance and microbial diversity between fecal microbes and the surrounding air microbes was significant (P<0.01), but the differences between air samples at different distances were not significant (P> 0.05).In addition, the microorganism OTUs differences in air samples at different distances were also analyzed using the QIIME software.The result shows that the OTUs of more than 25% samples contains are 2593, and the abundance significant differences OTUs are 27,but they account for only 1% in the total OTUs, so it indicates that the abundance difference is not significant in total samples at different distances, maybe due to the huge number of no significant differences OTUs, which covers up the significant differences OTUs.The 27 OTUs was observed, we found that there was no difference significance as the distance increases.From the level of OTUs, it was found out using the R software program that class A, B and C of microorganisms. Class A was pollution that those bacteria which numbers in the piggery are higher than that in the nearby of the piggery, and its content are decreasing with the distance gradient. Class B was potential pollution sources that the piggery than outside, but not with the content from the gradient of the microorganisms present. Class C is non-pollution including C1 type and C2 type,Cl is the microorganisms that piggery exit,but does not exist in the outside,C2 class is the microorganisms that piggery less than the outside.Meanwhile,the use of the R software program to find class D was those microorganisms which exists in feces,but undiscovered in the air,and class E was those microbes which Existed in the air but not found in feces.Class A microorganisms contains 149 OTUs,class B contains 1506 OTUs,class C1 contains 132 OTUs,class C2 contains 6689 OTUs,class D contains 969 OTUs, class E contains 436 OTUs. Analysis results of all kinds of microorganisms show that:1 A class microorganisms contains 149 OTUs, of which can spread to 10 meters have 149 OTUs,of which can spread to 20 meters have 15 OTUs,of which can spread to 50 meters has only one OTU, which indicated that the spread of microbial species and concentration process decreases rapidly as the distance increases, Class A microbial was identified to genus has 16 species,which have been reported as pathogenic microorganisms have 8 kinds,Only Prevotella can spread to 20 meters, No pathogenic microorganisms can spread to 50 meters, It is very necessary for pig farm design to maintain the health span,the health span should be at least 20 meters in terms of the gas source for bacterial infections.Under the experimental field conditions, bacterial infectious disease isolation is best to maintain health and safety of more than 50 meters span.In the process of farm planning, the distance of Isolated homes,living area,residential area around the farm and the first class road to piggery is greater than or equal to 50 meters in order to meet the needs of Preventing of bacterial diseases,2 Class D contains 969 OTUs, class E contains 436 OTUs, Class D microbial was identified to genus has 35 species, of which 18 species are reported as pathogenic microorganisms,There are four kinds of OTUs greater than or equal to 10 meters,they respectively are:Prevotella, Parabacteroides, Treponema, Bacteroides;Class E microbial identified to genus has 67 species,which have been reported as pathogenic microorganisms have 34 kinds,There are three kinds of OTUs greater than or equal to 10 meters, they are Streptococcus, Pseudomonas, Chryseobacterium respectively, these microbes can lead to animal or human infectious diseases, in addition, feces and air of microbes also has a large number of pathogenic microorganism,These analyzes indicate that disinfection of air environmental and control measures in large-scale pig farms has great significance for reducing the disease incidence of pigs.