field-deployable mesocosm culture of the larvae of crepidula fornicata

Odling av larver av Crepidula fornicata i laboratorie- och fältförhållanden

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.

Författare i fokus: Att överbrygga klyftan mellan fältobservationer och laboratoriemanipulationer i larvekologiforskning

Laboratorium och fältodling av larver av toffelsnäcka, Crepidula fornicata

I'm interested in how the development of marine larvae is influenced by multiple environmental factors and stressors and how the larval nervous system integrates information from the environment to regulate settlement of larvae, metamorphosis to benthic juveniles, and recruitment into adult populations. Larvae of some marine species like the Slipper Limpet, Crepidula Fornicata, are quite resilient to ocean acidification, but there are important interactions between acidification and other factors, like nutrition and salinity. We don't know very much about how larvae grow, develop, and behave in nature.Field mesocosm observations can fill in some of these gaps and suggest hypotheses to test with lab manipulations. We can begin to ask on neurobehavioral, developmental, and transcriptional levels how larvae deal with stressors that vary on short time scales in productive coastal environments that are especially susceptible to human impacts.

Mesokosmodling av larver av Crepidula fornicata som kan användas i fält

För att börja, använd en handhållen elektrisk sabelsåg för att skära fyra jämnt fördelade rektangulära öppningar i sidorna på en standard 7 gallon polyetenhink. Skär de utskurna bitarna på längden i 2 centimeter breda remsor med en bordssåg. Skär ut fyra paneler av nylonnät för att överlappa skopans utskurna öppningar.Fäst tillfälligt ena kanten av en nätpanel över långsidan av en utskärning med små fjäderklämmor. Limma nu fast kanten på nätpanelen på hinkens öppning med varmt lim. När den första kanten är fastsatt, limma ner de andra kanterna samtidigt som du behåller en toppspänning på panelen.Klipp sedan ner ändarna på de utskurna bitarna så att de passar över det limmade området på panelen. Tryck fast varje täckremsa med en generös mängd smältlim. Fäst slutligen änden av varje remsa med en blindnit i nylon som installerats från utsidan av skopan.Använd en rakhyvel för att trimma bort överflödigt lim och nät från kanterna på armeringsremsorna. Larverna av Crepidula Fornicata verkade växa snabbare i mesokosmen i förhållande till laboratoriekulturerna. Larverna metamorfoserar spontant mellan den femte och sjätte dagen.Nästan alla larver hade metamorfoserat sig på dag sju.

Research

JoVE Journal

Biology

Field-Deployable Mesocosm Culture of the Larvae of 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 ...

Collection of Larvae of Crepidula fornicata for Laboratory and Field Cultures

Samling av larver av Crepidula fornicata för laboratorie- och fältkulturer

Construction of Ventilated Cultures for Larvae of Crepidula fornicata

Konstruktion av ventilerade kulturer för larver av Crepidula fornicata

Setup and Routine Maintenance of Laboratory Cultures of Crepidula fornicata

Installation och rutinunderhåll av laboratoriekulturer av Crepidula fornicata