生物质炭对粪肥堆肥过程中抗生素抗性基因行为特征的影响
发布时间：2018-08-19 05:57
【摘要】：近年来,作为威胁人类健康的一种新型污染物“抗生素抗性基因(Antibiotic Resistance Gene, ARG)",其在畜禽粪肥中有较高的检出丰度,表明畜禽粪肥是抗生素抗性基因的一个重要蓄积库。本研究通过向三种不同的粪肥(猪粪肥、鸡粪肥和鸭粪肥)中添加不同种类的生物质炭(RSB：稻草生物质炭、CM：蘑菇渣生物质炭)进行好氧堆肥实验,研究了其对抗生素抗性基因(四环素类抗性基因、磺胺类抗性基因和氯霉素类抗性基因)、重金属(Cu、Zn和As)的影响。同时以鸡粪堆肥为例,用16S rRNA基因测序手段探究鸡粪肥堆肥过程中微生物群落结构的变化,并分析其与抗生素抗性基因的关系。结果表明：(1)堆肥前,抗生素抗性基因总丰度顺序为鸡粪肥鸭粪肥猪粪肥。在不添加生物质炭的情况下,抗生素抗性基因的平均去除率顺序为鸭粪肥堆肥(DM)猪粪肥堆肥(PM)鸡粪肥堆肥(CM),相对应的数值分别是2.58 logunits、2.19 log units and 0.80 log units。在猪粪肥和鸡粪肥堆肥过程中,总的来说,稻草生物质炭(RSB)的添加降低了抗生素抗性基因的平均去除率,而蘑菇渣生物质炭(MB)的添加增加了抗生素抗性基因的平均去除率。在鸭粪肥堆肥过程中,生物质炭的添加均降低了抗生素抗性基因的平均去除率。(2)在三种粪肥堆肥过程中,总量As和有效态As比例下降,生物质炭的添加进一步减少As的有效态比例。在鸡粪肥堆肥过程中,总量Cu、Zn下降,但有效态Cu、Zn比例上升,生物质炭的添加显著减少Cu、Zn的有效态比例(p0.05)。相关性分析结果还表明,与总量重和抗生素抗性基因的相关性相比,有效态重金属和抗生素抗性基因的相关性更明显。(3) 16S rRNA基因测序结果表明,鸡粪肥堆肥过程中优势菌群并未改变,均为厚壁菌门(Fimicutes)、拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)和放线菌门(Actinobacteria)。但优势菌群的比例发生了较大改变。粪肥堆肥过程中抗生素抗性基因丰度减少的原因可能是厚壁菌门Fimicutes含量的降低。和未添加生物质炭的堆肥相比,添加稻草生物质炭(RSB)的堆肥中属于厚壁菌门Fimicutes的三种细菌科大量增加,RDA分析结果表明这三种细菌科和抗生素抗性基因呈显著相关性(p0.05),所以稻草生物质炭(RSB)的添加会降低抗生素抗性基因去除率。此外,鸡粪肥堆肥过程中致病菌去除率的顺序和抗生素抗性基因去除率的顺序一致。
[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.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S141.4
[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.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S141.4
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