دورية أكاديمية
أعرض في EDS

eDNA-based monitoring of Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans with ddPCR in Luxembourg ponds: taking signals below the Limit of Detection (LOD) into account.

التفاصيل البيبلوغرافية
العنوان: eDNA-based monitoring of Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans with ddPCR in Luxembourg ponds: taking signals below the Limit of Detection (LOD) into account.
المؤلفون: Porco D; Musée national d'histoire naturelle du Luxembourg, 25, rue Münster, Luxembourg, L-2160, Luxembourg. david.porco.gm@gmail.com.; Fondation Faune Flore, 24, rue Münster, Luxembourg, L-2160, Luxembourg. david.porco.gm@gmail.com., Purnomo CA; Musée national d'histoire naturelle du Luxembourg, 25, rue Münster, Luxembourg, L-2160, Luxembourg., Glesener L; Naturschutzsyndikat SICONA, 12, rue de Capellen, L-8393 Olm, Luxembourg, Luxembourg., Proess R; Umweltplanungsbüro Ecotop, 45, Schlassuecht, L-7435 Hollenfels, Luxembourg, Luxembourg., Lippert S; Musée national d'histoire naturelle du Luxembourg, 25, rue Münster, Luxembourg, L-2160, Luxembourg., Jans K; Natur&ëmwelt Fondation Hëllef fir d'Natur, 5, Route de Luxembourg, L-1899, Kockelscheuer, Luxembourg., Colling G; Musée national d'histoire naturelle du Luxembourg, 25, rue Münster, Luxembourg, L-2160, Luxembourg.; Fondation Faune Flore, 24, rue Münster, Luxembourg, L-2160, Luxembourg., Schneider S; Musée national d'histoire naturelle du Luxembourg, 25, rue Münster, Luxembourg, L-2160, Luxembourg.; Naturschutzsyndikat SICONA, 12, rue de Capellen, L-8393 Olm, Luxembourg, Luxembourg., Stassen R; Biota.lu, 9a, Rue Principale, L-6990, Hostert, Luxembourg., Frantz AC; Musée national d'histoire naturelle du Luxembourg, 25, rue Münster, Luxembourg, L-2160, Luxembourg.; Fondation Faune Flore, 24, rue Münster, Luxembourg, L-2160, Luxembourg.
المصدر: BMC ecology and evolution [BMC Ecol Evol] 2024 Jan 05; Vol. 24 (1), pp. 4. Date of Electronic Publication: 2024 Jan 05.
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: BioMed Central Country of Publication: England NLM ID: 101775613 Publication Model: Electronic Cited Medium: Internet ISSN: 2730-7182 (Electronic) Linking ISSN: 27307182 NLM ISO Abbreviation: BMC Ecol Evol Subsets: MEDLINE
أسماء مطبوعة: Original Publication: London : BioMed Central, [2021]-
مواضيع طبية MeSH: Mycoses*/diagnosis , Mycoses*/microbiology , Chytridiomycota*/genetics , DNA, Environmental*, Animals ; Batrachochytrium/genetics ; Luxembourg ; Limit of Detection ; Ponds ; Amphibians/genetics ; Amphibians/microbiology ; Polymerase Chain Reaction/veterinary
مستخلص: Background: Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal) are two pathogenic fungi that are a significant threat to amphibian communities worldwide. European populations are strongly impacted and the monitoring of the presence and spread of these pathogens is crucial for efficient decision-making in conservation management.
Results: Here we proposed an environmental DNA (eDNA) monitoring of these two pathogenic agents through droplet digital PCR (ddPCR) based on water samples from 24 ponds in Luxembourg. In addition, amphibians were swabbed in eight of the targeted ponds in order to compare the two approaches at site-level detection. This study allowed the development of a new method taking below-Limit of Detection (LOD) results into account thanks to the statistical comparison of the frequencies of false positives in no template controls (NTC) and below-LOD results in technical replicates. In the eDNA-based approach, the use of this method led to an increase in Bd and Bsal detection of 28 and 50% respectively. In swabbing, this resulted in 8% more positive results for Bd. In some samples, the use of technical replicates allowed to recover above-LOD signals and increase Bd detection by 35 and 33% respectively for eDNA and swabbing, and Bsal detection by 25% for eDNA.
Conclusions: These results confirmed the usefulness of technical replicates to overcome high levels of stochasticity in very low concentration samples even for a highly sensitive technique such as ddPCR. In addition, it showed that below-LOD signals could be consistently recovered and the corresponding amplification events assigned either to positive or negative detection via the method developed here. This methodology might be particularly worth pursuing in pathogenic agents' detection as false negatives could have important adverse consequences. In total, 15 ponds were found positive for Bd and four for Bsal. This study reports the first record of Bsal in Luxembourg.
(© 2023. The Author(s).)
References: PLoS One. 2013 May 20;8(5):e63639. (PMID: 23703511)
Ecol Appl. 2021 Jul;31(5):e02342. (PMID: 33817953)
Biomol Detect Quantif. 2017 Apr 29;12:1-6. (PMID: 28702366)
Dis Aquat Organ. 2017 Mar 21;123(3):213-226. (PMID: 28322208)
J Wildl Dis. 2020 Jul;56(3):702-706. (PMID: 32243244)
PLoS One. 2012;7(4):e35038. (PMID: 22514705)
Dis Aquat Organ. 2022 Dec 15;152:99-108. (PMID: 36519681)
Nat Rev Microbiol. 2020 Jun;18(6):332-343. (PMID: 32099078)
Environ Sci Technol. 2015 May 5;49(9):5601-8. (PMID: 25850372)
Clin Biochem Rev. 2008 Aug;29 Suppl 1:S49-52. (PMID: 18852857)
Ecol Appl. 2023 Mar;33(2):e2785. (PMID: 36478292)
BMC Genomics. 2009 Mar 19;10:116. (PMID: 19298667)
R Soc Open Sci. 2017 Mar 1;4(3):160801. (PMID: 28405365)
Oecologia. 2018 Sep;188(1):319-330. (PMID: 29860635)
J Clin Microbiol. 2013 Dec;51(12):4173-7. (PMID: 24108616)
J Fungi (Basel). 2021 Mar 30;7(4):. (PMID: 33808405)
Science. 2019 Mar 29;363(6434):1459-1463. (PMID: 30923224)
Nature. 2012 Apr 11;484(7393):186-94. (PMID: 22498624)
Front Microbiol. 2017 Feb 02;8:108. (PMID: 28210243)
Nat Biotechnol. 2001 May;19(5):405. (PMID: 11328998)
PLoS One. 2014 Jan 21;9(1):e85999. (PMID: 24465831)
Dis Aquat Organ. 2019 Apr 11;134(1):15-24. (PMID: 32132269)
PLoS One. 2014 Sep 15;9(9):e106790. (PMID: 25222122)
J Wildl Dis. 2011 Apr;47(2):455-8. (PMID: 21441199)
Nature. 2017 Apr 19;544(7650):353-356. (PMID: 28425998)
Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):9236-41. (PMID: 10430926)
Dis Aquat Organ. 2017 Dec 19;127(1):75-79. (PMID: 29256431)
EFSA J. 2018 Apr 30;16(4):e05259. (PMID: 32625888)
Dis Aquat Organ. 2007 Jan 18;73(3):175-92. (PMID: 17330737)
Dis Aquat Organ. 2017 Sep 20;126(1):71-74. (PMID: 28930087)
PLoS One. 2022 Nov 16;17(11):e0275363. (PMID: 36383571)
Sci Total Environ. 2022 Dec 10;851(Pt 1):158143. (PMID: 35995149)
Dis Aquat Organ. 2004 Aug 9;60(2):141-8. (PMID: 15460858)
Fungal Biol. 2013 Feb;117(2):156-62. (PMID: 23452953)
PLoS One. 2013;8(4):e55943. (PMID: 23573183)
Anal Bioanal Chem. 2016 Nov;408(27):7583-7593. (PMID: 27558101)
Science. 2014 Oct 31;346(6209):630-1. (PMID: 25359973)
PLoS One. 2015 Mar 23;10(3):e0122763. (PMID: 25799582)
PLoS One. 2014 Oct 21;9(10):e111091. (PMID: 25333363)
فهرسة مساهمة: Keywords: Batrachochytrium dendrobatidis; Batrachochytrium salamandrivorans; Below-LOD signal; Luxembourg; Monitoring; Pathogens; ddPCR; eDNA
المشرفين على المادة: 0 (DNA, Environmental)
SCR Organism: Batrachochytrium salamandrivorans
تواريخ الأحداث: Date Created: 20240104 Date Completed: 20240108 Latest Revision: 20240108
رمز التحديث: 20240109
مُعرف محوري في PubMed: PMC10768104
DOI: 10.1186/s12862-023-02189-9
PMID: 38178008
قاعدة البيانات: MEDLINE
الوصف
تدمد:2730-7182
DOI:10.1186/s12862-023-02189-9