Influence of natural settlement cues on the metamorphosis of fiddler crab megalopae, Uca vocator (Decapoda: Ocypodidae)

Megalopae of many decapod crab species accelerate their development time to metamorphosis (TTM) when exposed to natural physical and/or chemical cues characteristic of the parental habitat. In the present study, the influence of natural settlement cues on the moulting rates and development TTM in megalopae of the fiddler crab Uca vocator was investigated. The effects of mud from different habitats (including well-preserved and degraded-polluted mangrove habitats) and conspecific adult 'odours' (seawater conditioned with crabs) on the induction of metamorphosis were compared with filtered pure seawater (control). 95 to 100% of the megalopae successfully metamorphosed to first juvenile crab stage in all treatments, including the control. However, the development TTM differed significantly among treatments. Settlement cues significantly shortened development, while moulting was delayed in their absence. The fact that megalopae responded to metamorphosis-stimulating cues originating from both adult and non-adult benthic habitats demonstrates that settlement in this species may occur in a wider range of habitats within the mangrove ecosystem, including impacted areas.Megalopas de muitas espécies de caranguejos decápodes aceleram seu período de desenvolvimento até a metamorfose (TOM) quando são expostas a estímulos naturais físicos e/ou químicos característicos do habitat parental. No presente estudo, a influência de estímulos naturais sobre as taxas de muda e sobre o PDM foi investigada nas megalopas do caranguejo violinistac. Os efeitos da (i) lama de diferentes habitats (incluindo habitats de um manguezal bem preservado e de um degradado e poluído) e (ii) 'odores' dos adultos conspecíficos (água do mar acondicionada com caranguejos) sobre a indução da metamorfose foram comparados com (iii) água do mar pura e filtrada (controle). 95 a 100% das megalopas realizaram a metamorfose com sucesso para o primeiro estágio de caranguejo juvenil em todos os tratamentos, incluindo o controle. No entanto, o PDM diferiu significativamente entre os tratamentos. Os estímulos encurtaram significativamente o desenvolvimento, enquanto que a muda foi retardada na ausência deles. O fato de que as megalopas responderam aos estímulos indutores da metamorfose oriundos de ambos os habitats bentônicos dos adultos e de habitats onde eles são ausentes, demonstra que o assentamento nesta espécie pode ocorrer em uma grande variedade de habitats dentro do ecossistema de manguezal, incluindo áreas impactadas

Expanding dispersal studies at hydrothermal vents through species identification of cryptic larval forms

Author Posting. © The Author(s), 2010. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Marine Biology 157 (2010): 1049-1062, doi:10.1007/s00227-009-1386-8.The rapid identification of hydrothermal vent-endemic larvae to the species level is a key limitation to understanding the dynamic processes that control the abundance and distribution of fauna in such a patchy and ephemeral environment. Many larval forms collected near vents, even those in groups such as gastropods that often form a morphologically distinct larval shell, have not been identified to species. We present a staged approach that combines morphological and molecular identification to optimize the capability, efficiency, and economy of identifying vent gastropod larvae from the northern East Pacific Rise (NEPR). With this approach, 15 new larval forms can be identified to species. A total of 33 of the 41 gastropod species inhabiting the NEPR, and 26 of the 27 gastropod species known to occur specifically in the 9° 50’ N region, can be identified to species. Morphological identification efforts are improved by new protoconch descriptions for Gorgoleptis spiralis, Lepetodrilus pustulosus, Nodopelta subnoda, and Echinopelta fistulosa. Even with these new morphological descriptions, the majority of lepetodrilids and peltospirids require molecular identification. Restriction fragment length polymorphism digests are presented as an economical method for identification of five species of Lepetodrilus and six species of peltospirids. The remaining unidentifiable specimens can be assigned to species by comparison to an expanded database of 18S ribosomal DNA. The broad utility of the staged approach was exemplified by the revelation of species-level variation in daily planktonic samples and the identification and characterization of egg capsules belonging to a conid gastropod Gymnobela sp. A. The improved molecular and morphological capabilities nearly double the number of species amenable to field studies of dispersal and population connectivity.Funding was provided by as Woods Hole Oceanographic Institution Deep Ocean Exploration Institute grant to L.M and S. Beaulieu, National Science Foundation grants OCE-0424953, OCE-9712233, and OCE-9619605 to L.M, OCE-0327261 to T.S., and OCE-0002458 to K. Von Damm, and a National Defense Science and Engineering Graduate fellowship to D.A

Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates

In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) as antitumor agent for the treatment of advanced soft tissue sarcoma. In this protocol, methods for bioactivity-guided isolation, purification and identification of secondary metabolites from marine invertebrates such as sponges, tunicates, soft corals and crinoids are discussed. To achieve this goal, solvent extraction of usually freeze-dried sample of marine organisms is performed. Next, the extract obtained is fractionated by liquid-liquid partitioning followed by various chromatographic separation techniques including thin layer chromatography, vacuum liquid chromatography, column chromatography (CC) and preparative high-performance reversed-phase liquid chromatography. Isolation of bioactive secondary metabolites is usually monitored by bioactivity assays, e.g., antioxidant (2,2-diphenyl-1-picryl hydrazyl) and cytotoxicity (microculture tetrazolium) activities that ultimately yield the active principles. Special care should be taken when performing isolation procedures adapted to the physical and chemical characteristics of the compounds isolated, particularly their lipo- or hydrophilic characters. Examples of isolation of compounds of different polarities from extracts of various marine invertebrates will be presented in this protocol. Structure elucidation is achieved using recent spectroscopic techniques, especially 2D NMR and mass spectrometry analysis