Study on Genetic Diversity of Poly-phosphate Accumulating Bacteria Isolated from Both Wastewater of Piggery and Catfish Ponds in the Mekong Delta

Le Quang Khoi, T. T. Ngon, C. N. Diệp
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Abstract

Poly-phosphate accumulating bacteria (PAB) is an important bacterial group that can take up large amounts of phosphate and accumulate as intracellular poly-phosphate, contributing to biological phosphorus removal in waste-water treatment. The study was conducted to analyze a genetic diversity of PAB community isolated from samples of water and sludge of intensive catfish ponds and effluent water and sludge obtained from piggery waste-water treated by bio-digesters in the Mekong Delta, Vietnam. In a total of 439 isolates, there are 48 isolates that have the potential of accumulating intracellular poly-phosphate shown main characters included shaped like a rods or short rod-shaped; motile, twitching movements or non-motile; expression of poly-phosphate electron dense granules within the cells; accumulated poly-phosphate from poly-phosphate kinase gene 1; the content of intracellular poly-phosphate varied from 10-9 to 10-12 mg/cell. Based on phylogenetic tree for partial 16S rRNA genes, there are 22 strains isolated from intensive catfish ponds included in four classes: Bacilli, Actinobacteria, Beta-proteobacteria, Gamma-proteobacteria, and 26 strains isolated from piggery waste-water treated by bio-digesters included in four classes: Bacilli, Actinobacteria, Alpha-proteobacteria, Gamma-proteobacteria. The process of analysis and comparison of genetic diversity of 48 isolates showed 16S rRNA sequences have nucleotide regions of high variability interspersed with nucleotide regions of low variability. The variation of o and Pi index formed the different types of haplotype in population of PAB, there were 25 haplotypes (genotypes) from 48 sequences. The difference of the structure of haplotypes formed a high diversity between them (Hd=0.91). The levels of haplotype diversity created many genotypes for the genetic variation and the ability to adapt to the environment in the evolutionary process of bacterial strains capable ofhigh accumulating poly-P. Comparison of genetic diversity between populations of bacteria isolated from two sampling places, the results showed that the nucleotide and haplotype diversity of the strains isolated in piggery waste-water treated biodigesters (Pi=0.16, h=14) were lower and the genetic conservation was higher than isolated strains in intensive catfish ponds (Pi=0.18, h=16). This is scientific basis for the selection of the sample sources for the isolation and selection of poly-phosphate accumulating strains.
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湄公河三角洲养猪场废水和鲶鱼池废水中聚磷酸盐积累菌的遗传多样性研究
聚磷酸盐积累菌(polyphosphate accumulation bacteria, PAB)是一种重要的细菌群,它能大量吸收磷酸盐,并在细胞内积累成聚磷酸盐,在废水处理中起生物除磷作用。该研究旨在分析从越南湄公河三角洲集约化鲶鱼池塘的水和污泥以及由生物沼气池处理的养猪场废水的废水和污泥中分离的PAB群落的遗传多样性。在439株分离株中,有48株具有细胞内聚积多磷酸的潜力,主要表现为棒状或短棒状;运动、抽搐或不运动;细胞内聚磷酸盐电子致密颗粒的表达;聚磷酸激酶基因1积累的聚磷酸;胞内多磷酸含量在10-9 ~ 10-12 mg/细胞之间变化。根据16S rRNA部分基因的系统进化树,从集约鲶鱼池中分离到22株菌株,分为芽胞杆菌、放线菌、β -变形菌、γ -变形菌4类;从沼气池处理的猪场废水中分离到26株菌株,分为芽胞杆菌、放线菌、α -变形菌、γ -变形菌4类。对48个分离株的遗传多样性分析比较表明,16S rRNA序列具有高变异性核苷酸区和低变异性核苷酸区穿插的特点。0和Pi指数的变化形成了PAB群体中不同类型的单倍型,48个序列中有25个单倍型(基因型)。单倍型结构的差异使其具有较高的多样性(Hd=0.91)。单倍型多样性水平为高聚磷积累菌株的遗传变异和环境适应能力创造了多种基因型。结果表明,猪场废水处理沼液中分离菌株的核苷酸和单倍型多样性(Pi=0.16, h=14)低于集约化鲶鱼池(Pi=0.18, h=16),遗传保守性高于集约化鲶鱼池(Pi=0.18, h=16)。这为多磷酸积累菌的分离和筛选提供了科学依据。
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