construction of ventilated cultures for larvae of crepidula fornicata

実験室および野外条件での Crepidula fornicata の幼虫の培養

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.

著者スポットライト:幼虫生態学研究における野外観察と実験室操作の間のギャップを埋める

スリッパカサガイ、Crepidula fornicataの幼虫の実験室および野外培養

Crepidula fornicataの幼虫のための換気培養物の構築

まず、瓶の蓋に5ミリメートルの穴を2つ開けます。10-32ナイロンナットを使用して、各穴にチューブバーブを取り付けます。チューブバーブのねじ山付き内側部分の開口部にシリコーンゴムアクアリウムシーラントを軽くたたきます。長さ20センチ、幅2ミリメートルのポリエチレンチューブを開口部に押し込みます。チューブの端にあるシーラントが硬化するのを待ってから、突き出た端を切り取り、チューブバーブの端と同じ高さになるようにします。

construction-of-ventilated-cultures-for-larvae-of-crepidula-fornicata

Research

JoVE Journal

Biology

Construction of Ventilated Cultures for Larvae of Crepidula fornicata

18 ビュー. まず、瓶の蓋に5ミリメートルの穴を2つ開けます。10-32ナイロンナットを使用して、各穴にチューブバーブを取り付けます。チューブバーブのねじ山付き内側部分の開口部にシリコーンゴムアクアリウムシーラントを軽くたたきます。長さ20センチ、幅2ミリメートルのポリエチレンチューブを開口部に押し込みます。チューブの端にあるシーラントが硬化するのを待...

ビデオ: Crepidula fornicataの幼虫のための換気培養物の構築

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 ...