含油废水酵母菌-SBR处理工艺及微生态群落解析研究
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摘要
含油废水来源广泛,进入水体后会造成严重的环境污染。常规的物理法和化学法对BOD去除能力较差,在实际应用中微生物法具有成本低、占地少、不需特殊设备、不会带来二次污染等优点,因而倍受青睐。酵母菌作为一种极为宝贵的微生物资源,由于它具有良好的耐酸、耐渗透压,成絮能力好等特点,且酵母菌能将大部分有机物转化成无毒且营养丰富的单细胞蛋白,具有很高的废水处理能力和饲料生产价值,因此广泛地应用于高浓度含油废水的处理。
本文建立了含油废水酵母菌-SBR法废水处理工艺,同时用PCR-DGGE以及荧光原位杂交(FISH)的分子生物学方法解析废水处理系统中酵母菌群落结构,并研究酵母菌的表面特性以了解优势菌定植机制,研究结果如下:
(1)10菌株酵母菌复合体系对含油废水有很好的处理效果。酵母菌SBR连续运行过程中,保证污泥负荷0.25 kg/(kg·d),曝气时间6 h,沉降2 h,N、P营养源的量按BOD:N:P=100:5:1投加的情况下,系统稳定运行,出水COD从3000 mg/L左右降低到200～350 mg/L ,COD去除率达到88%～93%,油含量由1500 mg/L左右降为45～100 mg/L,去除率为93%～96%,去除了含油废水中绝大部分的有机污染物。为使出水达到排放标准,使用活性污泥后处理,和酵母菌SBR组成复合系统,使得总COD的去除率高达97.5%,油的去除率达到99%以上,保证了出水的COD值在50～60 mg/L,油含量为10～20 mg/L,达到了工业污水综合排放标准(GB 8978-1996)。
(2)利用PCR-DGGE技术对酵母菌-SBR废水处理系统中酵母菌群落进行解析,证明菌株O2或W1、O3或G3、W2、未知菌株X1可以稳定存在于废水处理系统中,且O2或W1、未知菌株X1在系统中占有绝对优势。
(3)用FISH技术进一步检测对优势菌进行解析,证明在酵母菌SBR系统在长期运行过程中,相对于细菌,酵母菌占据着优势地位;实验所使用探针能与O2(Candida tropicalis)、G3、O3、W1、W2纯菌株成功杂交,但仅O2、O3的杂交信号发出的红色荧光较强。在酵母菌SBR系统连续运行过程中,探针所追踪的酵母菌在处理含油废水的过程中数量逐渐增多,到后期占有绝对的优势,说明样品中发出红色荧光的O2或O3占大多数,结合PCR-DGGE的结果,可知,在酵母菌-SBR系统中,O2和O3是绝对优势菌株,能够稳定存在于系统中。
(4)对酵母菌定植机制的研究表明,疏水性和絮凝性是菌株O2和O3成为优势菌的重要影响因素,而酵母菌株产乳化剂及乳化能力直接影响到废水中油脂的降解效果,但对于菌株是否成为含油废水处理中的优势菌,菌株乳化作用是的一个影响因素,但不是主要因素。
Oily wastewater is an widely variety sources.This wastewanter accessing to water will cause serious environmental pollution. Conventional physical and chemical methods on the BOD removal capacity is poor, but in the practical application, biological methods are acclaimed with its low cost, small footprint, no special equipment, not bringing about secondary pollution.
As a kind of extremely precious microorganism resources, yeast has good acidproof, osmotic pressure, and flocculation ability. Meanwhile, the yeast can convert the majority of non-toxic and organic nutrient-rich single-cell protein, which with high feeding value, therefore it widely applies in high concentration organic wastewater treatment.
This article establishes the yeast-SBR wastewater treatment system. Simultaneously, molecular biological methods of PCR-DGGE and FISH were applied to analyze he yeast community structure in wastewater treatment system. And then we study the surface characteristics of yeast to understand the mechanism of mechanism of dominant yeast colonization. The results as follows:
(1) 10 yeast strains on the oily wastewater system show good treatment effect. During the process of yeast SBR,ensure that sludge loading 0.25kg / (kg ? d), aeration time 6 h, settlement 2 h, N, P nutrition source volume is based on BOD: N: P = 100:5:1.It can make the system stable operation. When the yeast-SBR system stable, the system effluent COD is down to 200 ~ 350mg/L from 3000mg/L, COD removal rate reached 88% to 93%, oil content reduced to 45 ~ 100 mg/L from 1500mg/L, the removal rate is 93% to 96%, this system removed most of the organic pollutants in the oil-containing wastewater To meet the effluent discharge standards we use activated sludge post-processing, composed of yeast SBR as complex system. The complex system maked up 97.5% removal rate of COD and oil removal rate is up to 99%. This combination ensures the effluent COD values of 50~60 mg/L, the oil content of 10 ~ 20 mg/L, reaching to the Integrated Wastewater Discharge Standard (GB 8978-1996).
(2) Using PCR-DGGE technique to analysis yeast community in yeast-SBR wastewater treatment system .The study shows that the strain of O2 or W1, O3, or G3, W2, unknown strain X1 can exist in the wastewater treatment system, and O2, or W1, unknown strain X1 are the dominant strains in the system.
(3) For further testing, we use FISH technology to analysis the dominant strains. In the yeast-SBR system , yeasts occupy a favorable position as opposed to bacteria; probe can successful hybrid with the pure strains of O2 (Candida tropicalis), G3, O3, W1, W2 ,but only the hybridization signals of O2, O3 are strong. In the long run of yeast-SBR system, the number of yeast tracked by probe in oily wastewater treatment gradually increased. It indicates that the majority of strains in the sample were O2 or O3 with red fluorescence. Combined with the results of PCR-DGGE, we can see in the yeast-SBR system, O2 and O3 are absolutely dominant strains that can stably exist in the wastewater treatment system.
(4) On the Mechanism of yeast colonization, hydrophobicity and flocculation is important factors of O2 and O3 to become the dominant strains.Emulsifier production and emulsifying ability of yeast strains directly affected the lipid degradation in wastewater, but emulsifying ability is just a factor for strains to be dominant, not the significant one.
本文建立了含油废水酵母菌-SBR法废水处理工艺,同时用PCR-DGGE以及荧光原位杂交(FISH)的分子生物学方法解析废水处理系统中酵母菌群落结构,并研究酵母菌的表面特性以了解优势菌定植机制,研究结果如下:
(1)10菌株酵母菌复合体系对含油废水有很好的处理效果。酵母菌SBR连续运行过程中,保证污泥负荷0.25 kg/(kg·d),曝气时间6 h,沉降2 h,N、P营养源的量按BOD:N:P=100:5:1投加的情况下,系统稳定运行,出水COD从3000 mg/L左右降低到200～350 mg/L ,COD去除率达到88%～93%,油含量由1500 mg/L左右降为45～100 mg/L,去除率为93%～96%,去除了含油废水中绝大部分的有机污染物。为使出水达到排放标准,使用活性污泥后处理,和酵母菌SBR组成复合系统,使得总COD的去除率高达97.5%,油的去除率达到99%以上,保证了出水的COD值在50～60 mg/L,油含量为10～20 mg/L,达到了工业污水综合排放标准(GB 8978-1996)。
(2)利用PCR-DGGE技术对酵母菌-SBR废水处理系统中酵母菌群落进行解析,证明菌株O2或W1、O3或G3、W2、未知菌株X1可以稳定存在于废水处理系统中,且O2或W1、未知菌株X1在系统中占有绝对优势。
(3)用FISH技术进一步检测对优势菌进行解析,证明在酵母菌SBR系统在长期运行过程中,相对于细菌,酵母菌占据着优势地位;实验所使用探针能与O2(Candida tropicalis)、G3、O3、W1、W2纯菌株成功杂交,但仅O2、O3的杂交信号发出的红色荧光较强。在酵母菌SBR系统连续运行过程中,探针所追踪的酵母菌在处理含油废水的过程中数量逐渐增多,到后期占有绝对的优势,说明样品中发出红色荧光的O2或O3占大多数,结合PCR-DGGE的结果,可知,在酵母菌-SBR系统中,O2和O3是绝对优势菌株,能够稳定存在于系统中。
(4)对酵母菌定植机制的研究表明,疏水性和絮凝性是菌株O2和O3成为优势菌的重要影响因素,而酵母菌株产乳化剂及乳化能力直接影响到废水中油脂的降解效果,但对于菌株是否成为含油废水处理中的优势菌,菌株乳化作用是的一个影响因素,但不是主要因素。
Oily wastewater is an widely variety sources.This wastewanter accessing to water will cause serious environmental pollution. Conventional physical and chemical methods on the BOD removal capacity is poor, but in the practical application, biological methods are acclaimed with its low cost, small footprint, no special equipment, not bringing about secondary pollution.
As a kind of extremely precious microorganism resources, yeast has good acidproof, osmotic pressure, and flocculation ability. Meanwhile, the yeast can convert the majority of non-toxic and organic nutrient-rich single-cell protein, which with high feeding value, therefore it widely applies in high concentration organic wastewater treatment.
This article establishes the yeast-SBR wastewater treatment system. Simultaneously, molecular biological methods of PCR-DGGE and FISH were applied to analyze he yeast community structure in wastewater treatment system. And then we study the surface characteristics of yeast to understand the mechanism of mechanism of dominant yeast colonization. The results as follows:
(1) 10 yeast strains on the oily wastewater system show good treatment effect. During the process of yeast SBR,ensure that sludge loading 0.25kg / (kg ? d), aeration time 6 h, settlement 2 h, N, P nutrition source volume is based on BOD: N: P = 100:5:1.It can make the system stable operation. When the yeast-SBR system stable, the system effluent COD is down to 200 ~ 350mg/L from 3000mg/L, COD removal rate reached 88% to 93%, oil content reduced to 45 ~ 100 mg/L from 1500mg/L, the removal rate is 93% to 96%, this system removed most of the organic pollutants in the oil-containing wastewater To meet the effluent discharge standards we use activated sludge post-processing, composed of yeast SBR as complex system. The complex system maked up 97.5% removal rate of COD and oil removal rate is up to 99%. This combination ensures the effluent COD values of 50~60 mg/L, the oil content of 10 ~ 20 mg/L, reaching to the Integrated Wastewater Discharge Standard (GB 8978-1996).
(2) Using PCR-DGGE technique to analysis yeast community in yeast-SBR wastewater treatment system .The study shows that the strain of O2 or W1, O3, or G3, W2, unknown strain X1 can exist in the wastewater treatment system, and O2, or W1, unknown strain X1 are the dominant strains in the system.
(3) For further testing, we use FISH technology to analysis the dominant strains. In the yeast-SBR system , yeasts occupy a favorable position as opposed to bacteria; probe can successful hybrid with the pure strains of O2 (Candida tropicalis), G3, O3, W1, W2 ,but only the hybridization signals of O2, O3 are strong. In the long run of yeast-SBR system, the number of yeast tracked by probe in oily wastewater treatment gradually increased. It indicates that the majority of strains in the sample were O2 or O3 with red fluorescence. Combined with the results of PCR-DGGE, we can see in the yeast-SBR system, O2 and O3 are absolutely dominant strains that can stably exist in the wastewater treatment system.
(4) On the Mechanism of yeast colonization, hydrophobicity and flocculation is important factors of O2 and O3 to become the dominant strains.Emulsifier production and emulsifying ability of yeast strains directly affected the lipid degradation in wastewater, but emulsifying ability is just a factor for strains to be dominant, not the significant one.
引文
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