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浙江大学学报（农业与生命科学版）
Journal of Zhejiang University (Agriculture and Life Sciences)  2021, Vol. 47 Issue (1): 98-106    DOI: 10.3785/j.issn.1008-9209.2020.03.122
Resource utilization & environmental protection     
Characteristics of an endogenous compound microbial inoculant and its immobilization effect on wastewater treatment from pig feedlots
Xinxin YOU(),Sheng WANG,Linna DU()
Wenzhou Vocational College of Science and Technology, Wenzhou 325006, Zhejiang, China
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Abstract  

Taking the wastewater from large-scale pig feedlots as the research object and the original wastewater as the main nutrient source, an endogenous compound microbial inoculant was successfully prepared by the enrichment method, which was named as WKM, and its removal effects on chemical oxygen demand (COD) and ammonia nitrogen in the process of wastewater treatment from pig feedlots were analyzed and monitored. The results showed that the removal rates of COD and ammonia nitrogen in the pig feedlot wastewater (ZW) were 94.8% and 61.8%, respectively, after 3 d by adding the WKM. The high-throughput sequencing results indicated that the main microbial species of WKM belonged to Proteobacteria, Actinobacteria and Bacteroidetes at the phylum level. The main microbial species at the genus levelbelonged to Leucobacter, Castellaniella, Camelimonas, Moheibacter, Nitrosomonas, Cloacibacillus and Pusillimonas, of which the mass ratio was approximately 2∶2∶2∶2∶1.5∶1.5∶1. The results based on the clusters of orthologous groups of proteins (COG) analysis showed the abundances of amino acid transport and metabolism ([E]), energy production and conversion ([C]), carbohydrate transport and metabolism ([G]), inorganic ion transport and metabolism ([P]) and coenzyme transport and metabolism ([H]) were relatively high, which played an essential role in the conversion of organic substances such as proteins and lipids in the pig feedlot wastewater containing high concentrations of ammonia nitrogen and organic pollutants. Furthermore, the endogenous compound microbial inoculant immobilized with wheat bran showed a higher removal efficiency for COD in the pig feedlot wastewater (ZR), compared with the non-addition group (P＜0.01) and WKM group (P＜0.05). Besides, one-way analysis of variance showed that the immobilization ratio of 1∶50 (mass/volume) of wheat bran to compound microbial inoculant had the highest removal efficiency in COD (P＜0.05) among all the treatments, reaching to 89.4% in the pig feedlot wastewater (ZR). To sum up, the endogenous compound microbial inoculant immobilized with wheat bran has potential application prospect and practical value at wastewater treatment from pig feedlots.

Key words： wastewater from pig feedlots      endogenous compound microbial inoculant      high-throughput sequencing      wheat bran immobilization     
Received: 12 March 2020      Published: 09 March 2021
CLC:  X  
Corresponding Authors: Linna DU     E-mail: luckyxinyou@163.com;dlg1314@126.com
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Xinxin YOU
Sheng WANG
Linna DU
Cite this article:

Xinxin YOU,Sheng WANG,Linna DU. Characteristics of an endogenous compound microbial inoculant and its immobilization effect on wastewater treatment from pig feedlots. Journal of Zhejiang University (Agriculture and Life Sciences), 2021, 47(1): 98-106.

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http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2020.03.122     OR     http://www.zjujournals.com/agr/Y2021/V47/I1/98


一种内源性复合微生物菌剂的特性及其固定化对猪场粪污水的处理效果

以规模化猪场粪污水为研究对象，以污水原液为主要营养来源，通过富集内源性土著微生物得到复合微生物菌剂（WKM），并分析监测其在污水处理过程中对猪场粪污水化学需氧量（chemical oxygen demand, COD）和氨氮的削减效果。结果表明：投加WKM处理3 d后，猪场粪污水（ZW）中COD和氨氮去除率分别达94.8%和61.8%。通过高通量测序发现：复合微生物菌剂中主要微生物种类属于变形菌门（Proteobacteria）、放线菌门（Actinobacteria）和拟杆菌门（Bacteroidetes）；在属水平上，其主要的微生物种类属于白色杆菌属（Leucobacter）、卡斯特兰尼菌属（Castellaniella）、Camelimonas属、Moheibacter属、亚硝化单胞菌属（Nitrosomonas）、Cloacibacillus属和尼古丁降解菌属（Pusillimonas），丰度比例约为2∶2∶2∶2∶1.5∶1.5∶1。基于直系同源蛋白簇（clusters of orthologous groups of proteins, COG）功能分布预测表明：复合微生物菌剂WKM的微生物功能结构中，氨基酸转运和代谢（［E］）、能量产生与转化（［C］）、碳水化合物转运和代谢（［G］）、无机离子转运和代谢（［P］）、辅酶转运和代谢（［H］）的功能丰度相对较高，对含高浓度的氨氮和有机污染物的猪场粪污水中蛋白质及脂类等有机物质的转换起到了重要作用。在猪场粪污水（ZR）中，经麸皮固定化后的复合微生物菌剂对COD的去除率进一步提高，极显著高于无添加组（P＜0.01）和显著高于WKM组（P＜0.05）。通过固定化配比优化实验得出：按照质量体积比1∶50的比例混合的麸皮和复合微生物菌剂对COD的去除效果最佳，高达89.4%（P＜0.05）。综上所述，本研究制备获得的麸皮固定化复合微生物菌剂在猪场粪污水的除污处理中具有潜在的应用和实践价值。


关键词： 猪场粪污水,  内源性复合微生物菌剂,  高通量测序,  麸皮固定化 
Fig. 1 Removal efficiencies of COD and ammonia nitrogen in wastewater from pig feedlots treated by the endogenous compound microbial inoculant immo-bilized with wheat branWB: Wheat bran; WKM: Compound microbial inoculant; CK: Non-addition. m(WB)∶V(WKM)=1∶50. Single asterisk (*) and double asterisks (**) indicate significant and highly significant differences at the 0.05 and 0.01 probability levels, respectively.
Fig. 2 Classification and phylogenetic information map of WKM-1, WKM-2 and WKM-3The center of the figure is the evolutionary taxonomic tree of the top 100 species with high abundance, and the phylum of the top 20 species (marked with asterisks) are marked by different colors, and the sizes of the circle and the asterisk represent the level of abundance. The outer ring is a thermal ring, in which each ring is a sample (group) that corresponds to a color. The color intensity varies with species abundance.
Fig. 3 Heat map of the species abundance in WKM-1, WKM-2 and WKM-3
Fig. 4 Heat map of functional abundance in WKM-1, WKM-2 and WKM-3 based on clusters of orthologous groups of proteins (COG)
Fig. 5 Removal efficiencies of COD and ammonia nitrogen in wastewater from pig feedlots treated by the endogenous compound microbial inoculant immo-bilized with wheat bran of different ratiosSingle asterisk (*) indicates significant differences at the 0.05 probability level.
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