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CLC number: TU961; X172

On-line Access: 2020-03-17

Received: 2019-08-22

Revision Accepted: 2020-01-11

Crosschecked: 2020-02-19

Cited: 0

Clicked: 1526

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Yu Shao

https://orcid.org/0000-0003-2435-5618

Jing-qing Liu

https://orcid.org/0000-0001-5596-0365

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Journal of Zhejiang University SCIENCE A 2020 Vol.21 No.3 P.167-178

http://doi.org/10.1631/jzus.A1900416


Optimization of ultrasonic parameters for effective detachment of biofilm cells in an actual drinking water distribution system


Author(s):  Hong-xi Peng, Yu Shao, Yi-fu Zhang, Ruo-wei Wang, David Z. Zhu, Huan-yu Chen, Jing-qing Liu

Affiliation(s):  College of Civil Engineering and Architecture, Zhejiang University, Hangzhou 310058, China; more

Corresponding email(s):   shaoyu1979@zju.edu.cn, liujingqing@zju.edu.cn

Key Words:  Biofilm, Drinking water distribution system (DWDS), DNA yield, DNA quality, Ultrasound treatment (UST)


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Hong-xi Peng, Yu Shao, Yi-fu Zhang, Ruo-wei Wang, David Z. Zhu, Huan-yu Chen, Jing-qing Liu. Optimization of ultrasonic parameters for effective detachment of biofilm cells in an actual drinking water distribution system[J]. Journal of Zhejiang University Science A, 2020, 21(3): 167-178.

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publisher="Zhejiang University Press & Springer",
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Abstract: 
It is important to obtain a considerable quantity of DNA from oligotrophic environments such as a drinking water distribution system (DWDS) to study microbial communities by molecular biotechnology, and DNA yield is always one of the biggest problems when performing metagenomic sequencing on drinking water samples. To obtain as many microbes as possible, ultrasound has been widely used in cell detachment, but studies on the optimal ultrasonic parameters for biofilm in DWDS have rarely been seen. The effects of three ultrasonic parameters, including power, duration, and the number of ultrasound treatments (USTs) on the selected monoculture bacteria (Pelomonas sp.) biofilm were studied first. Then the optimal values of each ultrasonic parameter were initially determined. Based on these values, three levels of each ultrasonic parameter were selected, and then an orthogonal experiment was conducted to further study drinking water biofilm, and finally the optimal ultrasonic parameters for the effective separation of biofilm cells in DWDS were determined. The results showed that the optimal ultrasonic power, duration, and the number of USTs are 13 W, 1 min, and 15, respectively. A 20-min interval is needed between two USTs. The present optimal UST, which does not lose DNA quality, can increase the amount of extractable DNA by at least 4.78 times compared to samples without UST. This study provides a pretreatment methodology for extracting more and reliable DNA from biofilm in DWDS, and can better solve the problem of DNA collection in oligotrophic environments.

This journal manuscript by Pend and others describes a microbial study of DNA sampling from drinking water distribution pipes, and offers new insights on operational parameters of ultrasound techniques for reliable DNA harvesting. Using experiments, cPCR and other molecular biological methods, the authors conducted a statistical analysis of the results for optimal ultrasonic parameters when the biological sampling technique is used for sufficient extraction of the samples without compromising of the HPC sample integrity. The results are important to sampling and biological monitoring of drinking water systems, especially in compliance of biological standards in water systems. This can add to the understanding and contribute to standard setting. Thus, I think the presented research is valuable and of interests to journal's readers and to the scientific community in general.

促进饮用水供水管道生物膜脱落的超声处理参数优化

目的:从贫营养环境(如饮用水供水管道)中获取足量的DNA是用分子生物学的方法研究微生物群落的重要环节. 尽管如此,DNA量不足始终是对饮用水环境样品进行宏基因组测序时最大的问题之一. 为了获得尽可能多的微生物,本文采用超声处理进行微生物分离,并研究处理供水管道生物膜的最佳超声处理参数.
创新点:本研究提供了一种预处理方法,可以从实际供水管道生物膜中提取更多可靠的DNA,可以更好地解决贫营养环境中DNA的收集问题.
方法:1. 用挑选的细菌(Pelomonas sp.)形成的生物膜来初步研究三个超声参数的最佳水平,即功率、时间和超声处理次数; 2. 基于这些值,对每个超声参数选择三个水平,并对实验室培养的饮用水生物膜进行正交试验,以进一步确定能有效分离生物膜的最佳超声参数水平; 3. 使用研究所得的最优超声处理参数对从实际供水管道采集的生物膜进行处理,以验证其效果.
结论:1. 最佳超声功率、时间和次数分别为13 W、1 min和15次,且两次超声处理之间需要20 min的静置期; 2. 与没有进行超声处理的样品相比,研究所得的最优超声处理不会降低DNA质量,而且可以将提取的DNA量增加至少4.78倍.

关键词:生物膜; 饮用水供水管道; DNA量; DNA质量; 超声处理

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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