土霉素和磺胺二甲嘧啶對(duì)堆肥過(guò)程中微生物群落多樣性及抗性基因的影響
發(fā)布時(shí)間:2018-05-06 16:49
本文選題:土霉素 + 磺胺二甲嘧啶; 參考:《西北農(nóng)林科技大學(xué)》2015年碩士論文
【摘要】:畜禽養(yǎng)殖業(yè)中,抗生素作為防治疾病藥物被大量使用,但大部分抗生素會(huì)隨畜禽糞便排出。好氧高溫堆肥是畜禽糞便無(wú)害化處理和資源化利用的主要途徑之一,而畜禽糞便中殘留的抗生素會(huì)對(duì)好氧堆肥過(guò)程產(chǎn)生一定的影響。本試驗(yàn)選取畜禽糞便中殘留濃度較大、檢出率較高的土霉素(OTC)和磺胺二甲嘧啶(SM2)為研究對(duì)象,將添加不同濃度抗生素的畜禽糞便與小麥秸稈混合堆肥,采用酶活分析法、Biolog法、T-RFLP技術(shù)、高效液相色譜法及實(shí)時(shí)定量PCR技術(shù)等,研究了OTC和SM2對(duì)堆肥過(guò)程中堆料理化性質(zhì)、酶活性、微生物群落多樣性、抗生素殘留及抗性基因的影響。主要結(jié)果如下:1.OTC和SM2對(duì)堆料理化性質(zhì)及酶活性的影響經(jīng)過(guò)35 d的堆肥處理,各處理堆體溫度均在55℃以上保持了4 d,達(dá)到了無(wú)害化處理的標(biāo)準(zhǔn)。各處理GI指數(shù)均大于85%,C/N小于16,達(dá)到了腐熟標(biāo)準(zhǔn),但添加抗生素會(huì)在短期內(nèi)延緩堆肥進(jìn)程。高濃度(140 mg·kg-1 OTC+30 mg·kg-1 SM2)抗生素對(duì)脫氫酶活性呈先抑制后促進(jìn)的作用,而對(duì)脲酶活性的抑制作用則表現(xiàn)在堆肥后期。添加抗生素僅在堆肥前期對(duì)纖維素酶活有顯著抑制作用。2.OTC和SM2對(duì)堆肥過(guò)程中微生物群落多樣性的影響B(tài)iolog結(jié)果表明,在堆肥腐熟期高濃度處理AWCD值、Shannon指數(shù)和Simpson指數(shù)顯著大于CK處理。主成分分析表明,抗生素改變了堆肥過(guò)程中微生物群落功能多樣性,高濃度處理作用更為顯著,起分異作用的碳源主要為糖類(lèi)、羧酸類(lèi)及氨基酸類(lèi)。采用T-RFLP技術(shù)分析堆肥過(guò)程中細(xì)菌和真菌群落多樣性,結(jié)果表明抗生素能改變堆肥過(guò)程中微生物群落多樣性,且對(duì)真菌群落的影響更大。高濃度處理顯著提高了細(xì)菌群落多樣性而降低了真菌群落的多樣性。聚類(lèi)分析表明,添加抗生素使堆肥過(guò)程中細(xì)菌和真菌的群落結(jié)構(gòu)發(fā)生了變化,在堆肥高溫期,高濃度抗生素顯著改變了真菌群落結(jié)構(gòu)。3.堆肥中OTC和SM2的降解及抗性基因豐度的變化堆肥35 d后,低濃度(10 mg·kg-1 OTC+1 mg·kg-1 SM2)和高濃度(140 mg·kg-1 OTC+30 mg·kg-1 SM2)處理中OTC殘留率分別為2.8%和4.2%,SM2含量均減少到檢測(cè)限以下?股卦诙逊手械慕到夥弦患(jí)動(dòng)力學(xué)方程,半衰期為0.56 d~1.24 d。從堆肥各時(shí)期樣品中共檢出5種四環(huán)素類(lèi)抗性基因,2種磺胺類(lèi)抗性基因,1種I類(lèi)整合子,其中tet X基因豐度最大,而dfr A7最小。說(shuō)明好氧高溫堆肥是一種有效去除抗生素的方法,但不能徹底去除抗性基因,還需要進(jìn)一步處理還降低抗性基因?qū)Νh(huán)境帶來(lái)的風(fēng)險(xiǎn)。
[Abstract]:In livestock farming, antibiotics are widely used as disease control drugs, but most antibiotics are excreted with animal feces. Aerobic high temperature composting is one of the main ways of harmless disposal and resource utilization of livestock and poultry feces, and the antibiotic residues in animal manure will have a certain effect on aerobic composting process. In this experiment, oxytetracycline (OTC) and sulfadimethazine (SM2), which had higher residual concentration and higher detection rate, were selected as the research objects. The mixed compost of livestock and poultry manure with different concentrations of antibiotics and wheat straw was used as the research object. The effects of OTC and SM2 on physical and chemical properties, enzyme activity, microbial community diversity, antibiotic residues and resistance genes during composting were studied by using Biolog T-RFLP, high performance liquid chromatography (HPLC) and real-time quantitative PCR. The main results were as follows: 1. The effects of OTC and SM2 on the physicochemical properties and enzyme activity of the stacks reached the standard of innocuous treatment after 35 days of composting. The temperature of each treatment remained above 55 鈩,
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