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題名 | 以定量聚合酶連鎖反應監測活性污泥中具異營性硝化作用之Rhodococcus屬菌株=Detection of Nitrifying Rhodococcus spp. in Active Sludge by Quantitative Polymerase Chain Reaction |
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作者 | 程梅萍; 廖仁寶; 王西華; Cheng, Mei-ping; Liaw, Ren-bao; Wang, Hsi-hua; |
期刊 | 中國畜牧學會會誌 |
出版日期 | 19990300 |
卷期 | 28:1 1999.03[民88.03] |
頁次 | 頁57-67 |
分類號 | 445.2594 |
語文 | chi |
關鍵詞 | 定量聚合酶連鎖反應; 豬糞尿廢水; 硝化作用; Rhodococcus屬; Quantitaative polymerase chain reaction; Swine wastewater; Rhodococcus spp.; Nitrification; |
中文摘要 | 本試驗目的為建立定量聚合�t連銷反應(QPCR)以監測豬糞尿廢水處理 系統活性污泥中具異營性硝化作用之Rhodococcus spp.,藉以評估接種Rhodococcus spp. 對豬糞尿廢水硝化作用之影響。建立的內標準是將對Rhodococcus spp.專一 的16S rRNA基因選殖後,切去75bp,進行聚合�t連鎖反應後其產物與目標基因 產物可在瓊脂電泳膠片上分離出,並以影像分析系統定量。定量PCR的標準曲 線方程式為:log(N/NIC)=-5.6116+1.3059×log(No),R�揚�為0.99,偵 測極限為364個genome。將三段式豬糞尿處理系統之好氣處理槽廢水,調控其環 境條件,以利於異營性硝化作用之發生,並監測其中Rhodococcus spp.的生長。 在硝化作用方面,在爆氣槽中接種Rhodococcus spp.,在曝氣6小時內,硝化作 用較未接種者快,14小時後則差異不大。以QPCR法監測Rhodococcus spp.菌數, 發現未接種之起始活性污泥中已有 Rhodococcus spp. 存在,但所佔的比例較接種 者低,在曝氣早期接種者Rhodococcus spp.菌數較未接種者高,其後則差異漸減。 顯示生物處理時,調控環境因子至適合微生物生長與分解污染物較接種微生物重 要。 |
英文摘要 | The purpose of this study was to develop a quantitative polymerase chain reaction (QPCR) for quantification of Rhodococcus spp., which heterotrophically nitrifies in the swine wastewater treatment system, especially in the active sludge. By this way, the effect of inoculating Rhodococcus spp. on nitrification of wastewater was evaluated. The internal control for QPCR was constructed by cutting 75 bp of partially cloned 16S rRNA gene, which is specific to Rhodococcus spp. The PCR products of internal control and target gene could be separated by electrophoresis and quantified by densitometric scanning. The equation of standard curve of QPCR was:log (N/N)=15.6116+ 1.3059 × log(No), and the R square and detection limit were 0.99 and 364 genomes, respectively. The dissolved oxygen was increased in the aeration process of wastewater treatment system to benefit the nitrification, and the numbers of Rhodococcus spp. in the active sludge was measured by QPCR. The results showed inoculating Rhodococcus spp. increased the nitrification in the early stage, but made little difference after 14 hr of aeration. Therer were some intrinsic Rhodococcus species in the active sludge as detected by QPCR. In the early stage of aeration, the Rhodococcus spp. population of uninoculated sludge was less than the inoculated one, but the difference became small after 14 hr of incubation These results showed that conditional factors were more important than the primary factor (inoculation) during the process of biological treatment. |
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