Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Testing Different Membrane Filters for 16S rRNA Gene-Based Metabarcoding in Karstic Springs

Oana Teodora Moldovan
* ORCID logo ,
Andreea Baricz
* ,
Edina Szekeres
* ,
Marius Kenesz
,
Maria-Alexandra Hoaghia
,
Erika Andrea Levei
ORCID logo ,
Ionut Cornel Mirea
ORCID logo ,
Ruxandra Nastase-Bucur
ORCID logo ,
Traian Brad
ORCID logo ,
Horia Banciu
ORCID logo ,
Iulia Chiciudean
Version 1 : Received: 8 October 2020 / Approved: 12 October 2020 / Online: 12 October 2020 (17:39:14 CEST)

A peer-reviewed article of this Preprint also exists.

Moldovan, O.T.; Baricz, A.; Szekeres, E.; Kenesz, M.; Hoaghia, M.A.; Levei, E.A.; Mirea, I.C.; Năstase-Bucur, R.; Brad, T.; Chiciudean, I.; Banciu, H.L. Testing Different Membrane Filters for 16S rRNA Gene-Based Metabarcoding in Karstic Springs. Water 2020, 12, 3400. Moldovan, O.T.; Baricz, A.; Szekeres, E.; Kenesz, M.; Hoaghia, M.A.; Levei, E.A.; Mirea, I.C.; Năstase-Bucur, R.; Brad, T.; Chiciudean, I.; Banciu, H.L. Testing Different Membrane Filters for 16S rRNA Gene-Based Metabarcoding in Karstic Springs. Water 2020, 12, 3400.

Abstract

(1) Introduction: Karstic springs are used worldwide by rural communities as sources of fresh water for human use and livestock survival. In Romania, 1/3 of the population has no direct access to the public water supply. The present study is part of a country-wide project on developing simple, quick and cheap methods for seasonal environmental and microbiological monitoring of karstic springs used as drinking water supply by rural populations. Critical steps for the monitoring workflow consist in the evaluation of water quality and choosing of suitable membrane filters to efficiently capture environmental DNA for further microbial diversity estimation by 16S rRNA gene-based metabarcoding; (2) Methods: Several commercial membrane filters of different composition and pore sizes were tested on the water sampled from three karstic springs in Romania, followed by water chemistry and whole community 16S rRNA gene-based metabarcoding analysis; (3) Results: We found that the different types of applied membrane filters provide a varying recovery of diversities and abundances of both overall and pathogenic bacteria; and (4) Conclusions: The result of the experiment with different filters shows which are the best for amplicon-based metabarcoding monitoring of karst springs.

Keywords

spring water; karst; 16S rRNA gene; membrane filters; metabarcoding; pathogenic bacteria

Subject

Biology and Life Sciences, Anatomy and Physiology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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