摘要
为了筛选出微生物絮凝剂产生菌并探索该絮凝剂对猪场污水的絮凝效果,试验从收集的家畜粪便、毛发和血液样品中富集并筛选絮凝剂产生菌,采用形态学、生理生化反应和16S rDNA测序等方法对该分离菌进行分离鉴定,优化分离菌的产絮条件,并测定所产絮凝剂对猪场污水的絮凝效果。结果表明:筛选得到200多株絮凝剂产生菌,其中分离菌P71-1的絮凝活性最强,该菌在LB固体培养基上呈黄色,为圆形、革兰氏阳性菌,生化特性与标准菌株芬氏纤维化微细菌W6122相似,PCR扩增获得大小约为1 377 bp的单一条带,16S rDNA基因序列与GenBank中标准菌株芬氏纤维化微细菌W6122的同源性为99.71%,与已知的水生纤维化微细菌3 bp聚在一个进化分支上,综合判断该分离菌为纤维化微细菌。优化该菌的最佳培养基组成为葡萄糖8 g/L,酵母提取物4 g/L,K2HPO4 1 g/L,KH2PO4 1 g/L,MgCl2 0.5 g/L,种子液的接种量为1%;最佳培养条件为pH值7,培养温度30℃,培养时间24 h,摇床转数120 r/min。获得的絮凝剂MBF-P71对猪场污水最佳投放比为10 mL/L,助凝剂最佳投放比为5 mL/L,最佳絮凝效果为絮凝率93.63%、COD去除率88.32%、氨氮去除率40.63%、总氮去除率37.50%。说明微生物絮凝剂产生菌对猪场污水具有良好的絮凝活性,可作为猪场污水净化的潜在途径。
In order to screen for microbial flocculant producing strains, and explore the flocculating effect of the flocculant on pig farm effluent, in this study, flowulant producing strains were enriched and screened from livestock manure, hair and blood samples. Morphology, physiological and biochemical reactions and 16 S rDNA sequencing methods were used to isolate and identify the isolated strains. The flocculant production conditions were optimized, and the flocculant flocculation effect on pig farm sewage was measured. The results showed that over 200 flocculant producing strains were screened and isolated, with the strongest flocculant activity obtained from isolate strain P71-1, which was yellow and round on LB solid medium and gram staining was positive. Its biochemical characteristics were similar to standard strain Cellulosimicrobium funkei W6122. A single band of approximately 1 377 bp was obtained by PCR amplification. The 16 S rDNA gene sequence was 99.71% homologous with Cellulosimicrobiums W6122 from GenBank. The strain and the well-known strain of Cellulosimicrobium aquatile 3 bp gathered in an evolutionary branch and were identified as Cellulosimicrobiums through comprehensive analysis. The optimal medium composition for optimising this bacterium was glucose 8 g/L, yeast 4 g/L, K2HPO4 1 g/L, KH2PO4 1 g/L, MgCl2 0.5 g/L, and the seed solution inoculum is 1%. The optimum culture conditions were pH value 7, culture temperature 30 ℃, culture time 24 h, shaker rotation speed 120 r/min. The optimal ratio of MBF-P71 to pig farm wastewater was 10 mL/L, and the optimal ratio of coagulant was 5 mL/L. The optimal flocculation efficiency was 93.63%, COD removal rate 88.32%, removal rate of ammonia nitrogen was 40.63%, and the removal rate of total nitrogen was 37.50%. It indicated that the microbial flocculant-producing strains had good flocculation activity to pig farm sewage, and could be used as a potential method for pig farm sewage purification.
作者
章沙沙
徐健峰
柳增善
ZHANG Shasha;XU Jianfeng;LIU Zengshan(Key Laboratory of Zoonosis Research,Ministry of Education/College of Animal Medicine,Jilin University,Changchun 130062,China;Panjin Center for inspection and testing,Panjin 124010,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2021年第13期10-16,20,143,共9页
Heilongjiang Animal Science And veterinary Medicine
基金
“十三五”国家重点研发计划项目(2018YFD0500504)。
关键词
微生物絮凝剂
猪场污水
纤维化微细菌
分离
鉴定
絮凝活性
污水净化
microbial flocculant
pig farm sewage
Cellulosimicrobiums
screening
identification
flocculation activity
sewage purification
