An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juveniles
The pen shell Pinna nobilis is critically endangered due to a disease that has affected all open water populations since late 2016. Collection of early spats is considered a fundamental step for pen shell conservation. However, the identification between P. nobilis and P. rudis juveniles by morpholo...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-01-01
|
| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016122000899 |
| _version_ | 1811201301823881216 |
|---|---|
| author | Gaetano Catanese José Tena-Medialdea Marija Aleksandra Bel Dajković Milena Mičić José Rafaél García-March |
| author_facet | Gaetano Catanese José Tena-Medialdea Marija Aleksandra Bel Dajković Milena Mičić José Rafaél García-March |
| author_sort | Gaetano Catanese |
| collection | DOAJ |
| description | The pen shell Pinna nobilis is critically endangered due to a disease that has affected all open water populations since late 2016. Collection of early spats is considered a fundamental step for pen shell conservation. However, the identification between P. nobilis and P. rudis juveniles by morphology is a very difficult task. Furthermore, due to the small size of juveniles and high sensitivity to handling, the sampling for this purpose must not damage individuals. As a consequence, the application of molecular techniques for conservation strategies to identify threatened and endangered bivalve species is every day more and more necessary.In this study, we present the development of a multiplex-PCR procedure for the rapid identification of two Pinna species from eDNA water samples. Using species-specific primers, designed in the rRNA16S and rRNA12S mitochondrial genes, identification of species was obtained by cellular or extracellular DNA dissolved in water and differentiated based on the size of the amplified DNA fragments.• Development of a molecular multiplex-PCR procedure for the rapid identification of two Pinna species from eDNA water samples• Using specie-specific primers, the different species can be differentiated basing on the size of the amplified DNA fragments• This technique removes many of the limitations commonly associated with sampling of threatened and endangered juvenile bivalves for conservation strategies (sampling does not damage individuals). |
| first_indexed | 2024-04-12T02:18:40Z |
| format | Article |
| id | doaj.art-1ad870994fab428aa022bc87aab57d32 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-04-12T02:18:40Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-1ad870994fab428aa022bc87aab57d322022-12-22T03:52:10ZengElsevierMethodsX2215-01612022-01-019101708An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juvenilesGaetano Catanese0José Tena-Medialdea1Marija Aleksandra Bel Dajković2Milena Mičić3José Rafaél García-March4Laboratorio de Investigaciones Marinas y Acuicultura (LIMIA) - Govern de les Illes Balears, Av. Gabriel Roca 69, 07157 Port d'Andratx (Balearic Islands) Spain; INAGEA-UIB, Carr. de Valldemossa, km 7.5, 07122 Palma, (Balearic Islands) Spain; Corresponding author.Instituto de Investigación en Medio Ambiente y Ciencia Marina (IMEDMAR-UCV). Universidad Católica de Valencia. Avenida Puerto Pesquero s/n, 03710 Calpe (Spain)Aquarium Pula d.o.o., Ulica Verudella 33, 52100 Pula (Croatia)Aquarium Pula d.o.o., Ulica Verudella 33, 52100 Pula (Croatia)Instituto de Investigación en Medio Ambiente y Ciencia Marina (IMEDMAR-UCV). Universidad Católica de Valencia. Avenida Puerto Pesquero s/n, 03710 Calpe (Spain)The pen shell Pinna nobilis is critically endangered due to a disease that has affected all open water populations since late 2016. Collection of early spats is considered a fundamental step for pen shell conservation. However, the identification between P. nobilis and P. rudis juveniles by morphology is a very difficult task. Furthermore, due to the small size of juveniles and high sensitivity to handling, the sampling for this purpose must not damage individuals. As a consequence, the application of molecular techniques for conservation strategies to identify threatened and endangered bivalve species is every day more and more necessary.In this study, we present the development of a multiplex-PCR procedure for the rapid identification of two Pinna species from eDNA water samples. Using species-specific primers, designed in the rRNA16S and rRNA12S mitochondrial genes, identification of species was obtained by cellular or extracellular DNA dissolved in water and differentiated based on the size of the amplified DNA fragments.• Development of a molecular multiplex-PCR procedure for the rapid identification of two Pinna species from eDNA water samples• Using specie-specific primers, the different species can be differentiated basing on the size of the amplified DNA fragments• This technique removes many of the limitations commonly associated with sampling of threatened and endangered juvenile bivalves for conservation strategies (sampling does not damage individuals).http://www.sciencedirect.com/science/article/pii/S2215016122000899Pen shelleDNAMolecular biologyMultiplex-PCR |
| spellingShingle | Gaetano Catanese José Tena-Medialdea Marija Aleksandra Bel Dajković Milena Mičić José Rafaél García-March An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juveniles MethodsX Pen shell eDNA Molecular biology Multiplex-PCR |
| title | An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juveniles |
| title_full | An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juveniles |
| title_fullStr | An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juveniles |
| title_full_unstemmed | An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juveniles |
| title_short | An incubation water eDNA method for a non-destructive rapid molecular identification of Pinna nobilis and Pinna rudis bivalve juveniles |
| title_sort | incubation water edna method for a non destructive rapid molecular identification of pinna nobilis and pinna rudis bivalve juveniles |
| topic | Pen shell eDNA Molecular biology Multiplex-PCR |
| url | http://www.sciencedirect.com/science/article/pii/S2215016122000899 |
| work_keys_str_mv | AT gaetanocatanese anincubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT josetenamedialdea anincubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT marijaaleksandrabeldajkovic anincubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT milenamicic anincubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT joserafaelgarciamarch anincubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT gaetanocatanese incubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT josetenamedialdea incubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT marijaaleksandrabeldajkovic incubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT milenamicic incubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles AT joserafaelgarciamarch incubationwaterednamethodforanondestructiverapidmolecularidentificationofpinnanobilisandpinnarudisbivalvejuveniles |