【摘要】：近年來,作為威脅人類健康的一種新型污染物“抗生素抗性基因(Antibiotic Resistance Gene, ARG)",其在畜禽糞肥中有較高的檢出豐度,表明畜禽糞肥是抗生素抗性基因的一個重要蓄積庫。本研究通過向三種不同的糞肥(豬糞肥、雞糞肥和鴨糞肥)中添加不同種類的生物質(zhì)炭(RSB：稻草生物質(zhì)炭、CM：蘑菇渣生物質(zhì)炭)進行好氧堆肥實驗,研究了其對抗生素抗性基因(四環(huán)素類抗性基因、磺胺類抗性基因和氯霉素類抗性基因)、重金屬(Cu、Zn和As)的影響。同時以雞糞堆肥為例,用16S rRNA基因測序手段探究雞糞肥堆肥過程中微生物群落結(jié)構(gòu)的變化,并分析其與抗生素抗性基因的關(guān)系。結(jié)果表明：(1)堆肥前,抗生素抗性基因總豐度順序為雞糞肥鴨糞肥豬糞肥。在不添加生物質(zhì)炭的情況下,抗生素抗性基因的平均去除率順序為鴨糞肥堆肥(DM)豬糞肥堆肥(PM)雞糞肥堆肥(CM),相對應(yīng)的數(shù)值分別是2.58 logunits、2.19 log units and 0.80 log units。在豬糞肥和雞糞肥堆肥過程中,總的來說,稻草生物質(zhì)炭(RSB)的添加降低了抗生素抗性基因的平均去除率,而蘑菇渣生物質(zhì)炭(MB)的添加增加了抗生素抗性基因的平均去除率。在鴨糞肥堆肥過程中,生物質(zhì)炭的添加均降低了抗生素抗性基因的平均去除率。(2)在三種糞肥堆肥過程中,總量As和有效態(tài)As比例下降,生物質(zhì)炭的添加進一步減少As的有效態(tài)比例。在雞糞肥堆肥過程中,總量Cu、Zn下降,但有效態(tài)Cu、Zn比例上升,生物質(zhì)炭的添加顯著減少Cu、Zn的有效態(tài)比例(p0.05)。相關(guān)性分析結(jié)果還表明,與總量重和抗生素抗性基因的相關(guān)性相比,有效態(tài)重金屬和抗生素抗性基因的相關(guān)性更明顯。(3) 16S rRNA基因測序結(jié)果表明,雞糞肥堆肥過程中優(yōu)勢菌群并未改變,均為厚壁菌門(Fimicutes)、擬桿菌門(Bacteroidetes)、變形菌門(Proteobacteria)和放線菌門(Actinobacteria)。但優(yōu)勢菌群的比例發(fā)生了較大改變。糞肥堆肥過程中抗生素抗性基因豐度減少的原因可能是厚壁菌門Fimicutes含量的降低。和未添加生物質(zhì)炭的堆肥相比,添加稻草生物質(zhì)炭(RSB)的堆肥中屬于厚壁菌門Fimicutes的三種細菌科大量增加,RDA分析結(jié)果表明這三種細菌科和抗生素抗性基因呈顯著相關(guān)性(p0.05),所以稻草生物質(zhì)炭(RSB)的添加會降低抗生素抗性基因去除率。此外,雞糞肥堆肥過程中致病菌去除率的順序和抗生素抗性基因去除率的順序一致。
[Abstract]:In recent years, as a new pollutant "antibiotic resistance gene (Antibiotic Resistance Gene, ARG)", which is a new pollutant threatening human health, it has a high detection abundance in livestock manure, which indicates that livestock manure is an important accumulation of antibiotic resistance genes. In this study, aerobic composting experiments were carried out by adding different kinds of biomass carbon (RSB: straw biomass carbon CMC: mushroom residue biomass carbon) to three different manure (pig manure, chicken manure and duck manure). Its effects on antibiotic resistance genes (tetracycline resistance genes, sulfonamides resistance genes and chloramphenicol resistance genes) and heavy metals (Cu, Zn and as) were studied. At the same time, taking chicken manure compost as an example, 16s rRNA gene sequencing was used to study the changes of microbial community structure in chicken manure compost, and the relationship between the microbial community structure and antibiotic resistance gene was analyzed. The results showed that: (1) before composting, the sequence of total abundance of antibiotic resistance genes was chicken manure, duck manure, pig manure. Without adding biomass carbon, the average removal rate of antibiotic resistance gene was 2.58 logunits2.19 log units and 0.80 log unitsof duck manure compost, (DM) pig manure compost, (PM) chicken manure compost, (CM),. In the process of pig manure and chicken manure composting, in general, the addition of straw biomass carbon (RSB) reduced the average removal rate of antibiotic resistance gene, while the addition of mushroom residue biomass carbon (MB) increased the average removal rate of antibiotic resistance gene. In the process of duck manure composting, the addition of biomass carbon decreased the average removal rate of antibiotic resistance genes. (2) during the three kinds of manure composting, the ratio of total as and available as decreased. The addition of biomass carbon further reduced the ratio of effective state of as. During the composting of chicken manure, the total amount of Cu ~ (2 +) Zn decreased, but the ratio of available Cu ~ (2 +) Zn increased, and the proportion of available Cu ~ (2 +) Zn decreased significantly (p0.05) by the addition of biomass carbon. The results of correlation analysis also showed that the correlation between available heavy metals and antibiotic resistance genes was more obvious than that of total weight and antibiotic resistance genes. (3) 16s rRNA gene sequencing showed that, During the composting process of chicken manure compost, the dominant microflora did not change, all of them were (Fimicutes), (Bacteroidetes), (Bacteroidetes), (Proteobacteria), and actinomycetes, (Actinobacteria). But the proportion of dominant bacteria changed greatly. The decrease in the abundance of antibiotic resistance genes during manure composting may be due to the decrease of Fimicutes content in the phylum thuringiensis. Compared with compost without biomass carbon, The results of (RSB) analysis showed that there was a significant correlation between the three bacterial families and antibiotic resistance genes (p0.05), so the addition of (RSB) in straw biomass charcoal compost showed a significant correlation (p0.05). It reduces the removal rate of antibiotic resistance genes. In addition, the sequence of the removal rate of pathogenic bacteria in chicken manure compost was the same as that of antibiotic resistance gene.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S141.4

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