field-deployable mesocosm culture of the larvae of crepidula fornicata

Kultivierung von Larven von Crepidula fornicata unter Labor- und Freilandbedingungen

The slipper limpet, Crepidula fornicata (Gastropoda: Calyptraeidae), is well-represented in current and historical research literature because of its utility as a developmental model and because of its widespread impacts as an invasive species. It served as a foundational example of spiralian development in the classic age of experimental embryology1 and has experienced a rebirth of interest with the application of modern imaging and genomic tools to dissect mechanisms of lophotrochozoan early development2,3. At the other end of its life history, other investigations have focused on the impacts of adult populations of this ecosystem engineer in temperate coastal marine environments far removed from its original distribution in eastern North America4,5. In between embryo and adult, the veliger larvae of this species have been subjects of numerous studies of larval development and ecology, especially of factors influencing growth and acquisition of competence for metamorphosis, the internal and external cues mediating larval settlement, and the effects of larval experience on juvenile performance6,7,8,9,10,11. Recent studies have revealed the resilience of larvae and juveniles of C. fornicata to ocean acidification, yet another avenue for productive research use of this animal12,13,14,15,16.
An advantage of C. fornicata for studies of marine larval biology is that it is relatively easy to grow in the laboratory in natural or artificial seawater on a unialgal diet of the flagellate Isochrysis galbana. Culture methods have been detailed by the author in an earlier methods-focused print publication17. The reasons for the present contribution are twofold. First, the routine physical maneuvers involved in establishing and caring for cultures are conceptually very simple but difficult to perform correctly without hands-on or video demonstration. Second, two variations on previously described culture methods are described that are especially suited to laboratory and field studies of responses to environmental stressors such as ocean acidification, eutrophication, and oxygen depletion. The first of these is a low-volume (800 mL) culture system suited for manipulation of pH and dissolved oxygen in seawater via small volumes of bubbled gases, and the second is a larger volume (15 L) mesocosm system that can be placed in the field and that allows free exchange of ambient seawater.

Autor im Rampenlicht: Überbrückung der Lücke zwischen Feldbeobachtungen und Labormanipulationen in der Larvenökologieforschung

Labor- und Feldkultur von Larven des Pantoffelzuwandes, Crepidula fornicata

Feldeinsetzbare Mesokosmenkultur der Larven von Crepidula fornicata

Schneiden Sie zunächst mit einer elektrischen Handsäge vier gleichmäßig verteilte rechteckige Öffnungen in die Seiten eines Standard-7-Gallonen-Polyethyleneimers. Schneiden Sie mit einer Tischkreissäge die ausgeschnittenen Stücke der Länge nach in 2 Zentimeter breite Streifen. Schneiden Sie vier Paneele aus Nylonnetz aus, um die Ausschnittöffnungen des Eimers zu überlappen.Befestigen Sie vorübergehend eine Kante eines Gitterfeldes über der langen Kante eines Ausschnitts mit kleinen Federklemmen. Kleben Sie nun den Rand der Gitterplatte mit Heißkleber an die Eimeröffnung. Sobald die erste Kante befestigt ist, kleben Sie die anderen Kanten fest, während Sie eine obere Spannung auf der Platte beibehalten.Schneiden Sie anschließend die Enden der ausgeschnittenen Stücke so ab, dass sie über die geklebte Fläche der Platte passen. Drücken Sie jeden Abdeckstreifen mit einer großzügigen Menge Schmelzkleber an Ort und Stelle. Befestigen Sie abschließend das Ende jedes Streifens mit einem Nylon-Blindniet, der von außerhalb des Eimers angebracht wird.Verwende einen Rasierer, um den überschüssigen Kleber und das Netz von den Rändern der Verstärkungsstreifen abzuschneiden. Die Larven von Crepidula Fornicata schienen im Mesokosmen im Vergleich zu den Laborkulturen schneller zu wachsen. Die Larven metamorphosieren sich spontan zwischen dem fünften und sechsten Tag.Fast alle Larven hatten sich am siebten Tag metamorphosiert.

field-deployable-mesocosm-culture-of-the-larvae-of-crepidula-fornicata

Research

JoVE Journal

Biology

Field-Deployable Mesocosm Culture of the Larvae of Crepidula fornicata

79 Aufrufe. Schneiden Sie zunächst mit einer elektrischen Handsäge vier gleichmäßig verteilte rechteckige Öffnungen in die Seiten eines Standard-7-Gallonen-Polyethyleneimers. Schneiden Sie mit einer Tischkreissäge die ausgeschnittenen Stücke der Länge nach in 2 Zentimeter breite Streifen. Schneiden Sie vier Paneele aus Nylonnetz aus, um die Ausschnittöffnungen des Eimers zu überlappen.Befestigen Sie vorübergehend eine Kante eines Gitterfeldes über der langen Kante eines Ausschnitts mit...

Serie: Feldeinsetzbare Mesokosmenkultur der Larven von Crepidula fornicata

Laboratory and Field Culture of Larvae of The Slipper Limpet, Crepidula fornicata

The calyptraeid gastropod mollusk, Crepidula fornicata, has been widely used for studies of larval developmental biology, physiology, and ecology. Brooded veliger larvae of this species were collected by siphoning onto a sieve after natural release by adults, distributed into the culture at a density of 200/L, and fed with Isochrysis galbana (strain T-ISO) at 1 x 105 cells/mL. Shell growth and acquisition of competence for metamorphosis were documented for sibling larvae reared in ventilated 800 mL cultures designed for equilibration to ambient air or to defined atmospheric gas mixtures. Contrasting with these laboratory culture conditions; growth and competence data were also collected for larvae reared in a 15 L flow-through ambient seawater mesocosm located in a field population of reproductive adults. Growth rates and timing of metamorphic competence in the laboratory cultures were similar to those reported in previously published studies. Larvae reared in the field mesocosm grew much faster and metamorphosed sooner than reported for any laboratory studies. Together, these methods are suited for exploring larval development under predetermined controlled conditions in the laboratory as well as under naturally occurring conditions in the field.

The paper presents methods for larval culture of the gastropod Crepidula fornicata in a small-volume laboratory-scale system and in an ambient-seawater mesocosm system that can be deployed in the field.

The calyptraeid gastropod mollusk, Crepidula fornicata, has been widely used for studies of larval developmental biology, physiology, and ecology. Brooded ...