Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays

Abstract The invasive American bullfrog (Lithobates catesbeianus) imperils freshwater biodiversity worldwide. Effective management hinges on early detection of incipient invasions and subsequent rapid response, as established populations are extremely difficult to eradicate. Although environmental D...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Teun Everts, David Halfmaerten, Sabrina Neyrinck, Nico De Regge, Hans Jacquemyn, Rein Brys
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3244cecf9d9a4550b8cafe6651474d58
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:3244cecf9d9a4550b8cafe6651474d58
record_format	dspace
spelling	oai:doaj.org-article:3244cecf9d9a4550b8cafe6651474d582021-12-02T16:53:11ZAccurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays10.1038/s41598-021-90771-w2045-2322https://doaj.org/article/3244cecf9d9a4550b8cafe6651474d582021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90771-whttps://doaj.org/toc/2045-2322Abstract The invasive American bullfrog (Lithobates catesbeianus) imperils freshwater biodiversity worldwide. Effective management hinges on early detection of incipient invasions and subsequent rapid response, as established populations are extremely difficult to eradicate. Although environmental DNA (eDNA) detection methods provide a highly sensitive alternative to conventional surveillance techniques, extensive testing is imperative to generate reliable output. Here, we tested and compared the performance of two primer/probe assays to detect and quantify the abundance of bullfrogs in Western Europe in silico and in situ using digital droplet PCR (ddPCR). Although both assays proved to be equally target-specific and sensitive, one outperformed the other in ddPCR detection resolution (i.e., distinguishing groups of target-positive and target-negative droplets), and hence was selected for further analyses. Mesocosm experiments revealed that tadpole abundance and biomass explained 99% of the variation in eDNA concentration. Because per individual eDNA emission rates did not differ significantly among tadpoles and juveniles, and adults mostly reside out of the water, eDNA concentration can be used as an approximation of local bullfrog abundance in natural populations. Seasonal eDNA patterns in three colonized ponds showed parallel fluctuations in bullfrog eDNA concentration. An increase in eDNA concentration was detected in spring, followed by a strong peak coinciding with the breeding season (August, September or October), and continuously low eDNA concentrations during winter. With this study, we report the validation process required for appropriately implementing eDNA barcoding analyses in lentic systems. We demonstrate that this technique can serve as a solid and reliable tool to detect the early stages of bullfrog invasions and to quantify temporal changes in abundance that will be useful in coordinating large-scale bullfrog eradication programs and evaluating their efficiency.Teun EvertsDavid HalfmaertenSabrina NeyrinckNico De ReggeHans JacquemynRein BrysNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Teun Everts David Halfmaerten Sabrina Neyrinck Nico De Regge Hans Jacquemyn Rein Brys Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays
description	Abstract The invasive American bullfrog (Lithobates catesbeianus) imperils freshwater biodiversity worldwide. Effective management hinges on early detection of incipient invasions and subsequent rapid response, as established populations are extremely difficult to eradicate. Although environmental DNA (eDNA) detection methods provide a highly sensitive alternative to conventional surveillance techniques, extensive testing is imperative to generate reliable output. Here, we tested and compared the performance of two primer/probe assays to detect and quantify the abundance of bullfrogs in Western Europe in silico and in situ using digital droplet PCR (ddPCR). Although both assays proved to be equally target-specific and sensitive, one outperformed the other in ddPCR detection resolution (i.e., distinguishing groups of target-positive and target-negative droplets), and hence was selected for further analyses. Mesocosm experiments revealed that tadpole abundance and biomass explained 99% of the variation in eDNA concentration. Because per individual eDNA emission rates did not differ significantly among tadpoles and juveniles, and adults mostly reside out of the water, eDNA concentration can be used as an approximation of local bullfrog abundance in natural populations. Seasonal eDNA patterns in three colonized ponds showed parallel fluctuations in bullfrog eDNA concentration. An increase in eDNA concentration was detected in spring, followed by a strong peak coinciding with the breeding season (August, September or October), and continuously low eDNA concentrations during winter. With this study, we report the validation process required for appropriately implementing eDNA barcoding analyses in lentic systems. We demonstrate that this technique can serve as a solid and reliable tool to detect the early stages of bullfrog invasions and to quantify temporal changes in abundance that will be useful in coordinating large-scale bullfrog eradication programs and evaluating their efficiency.
format	article
author	Teun Everts David Halfmaerten Sabrina Neyrinck Nico De Regge Hans Jacquemyn Rein Brys
author_facet	Teun Everts David Halfmaerten Sabrina Neyrinck Nico De Regge Hans Jacquemyn Rein Brys
author_sort	Teun Everts
title	Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays
title_short	Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays
title_full	Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays
title_fullStr	Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays
title_full_unstemmed	Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays
title_sort	accurate detection and quantification of seasonal abundance of american bullfrog (lithobates catesbeianus) using ddpcr edna assays
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3244cecf9d9a4550b8cafe6651474d58
work_keys_str_mv	AT teuneverts accuratedetectionandquantificationofseasonalabundanceofamericanbullfroglithobatescatesbeianususingddpcrednaassays AT davidhalfmaerten accuratedetectionandquantificationofseasonalabundanceofamericanbullfroglithobatescatesbeianususingddpcrednaassays AT sabrinaneyrinck accuratedetectionandquantificationofseasonalabundanceofamericanbullfroglithobatescatesbeianususingddpcrednaassays AT nicoderegge accuratedetectionandquantificationofseasonalabundanceofamericanbullfroglithobatescatesbeianususingddpcrednaassays AT hansjacquemyn accuratedetectionandquantificationofseasonalabundanceofamericanbullfroglithobatescatesbeianususingddpcrednaassays AT reinbrys accuratedetectionandquantificationofseasonalabundanceofamericanbullfroglithobatescatesbeianususingddpcrednaassays
_version_	1718382849212547072

Accurate detection and quantification of seasonal abundance of American bullfrog (Lithobates catesbeianus) using ddPCR eDNA assays

Ejemplares similares