Sven Zea

Does the odor from sponges of the genus Ircinia protect them from fish predators

Caribbean sponges of the genus Ircinia contain high concentrations of linear furanosesterterpene tetronic acids (FTAs) and produce and exude low-molecular-weight volatile compounds (e.g., dimethyl sulfide, methyl isocyanide, methyl isothiocyanate) that give these sponges their characteristic unpleasant garlic odor. It has recently been suggested that FTAs are unlikely to function as antipredatory chemical defenses, and this function may instead be attributed to bioactive volatiles. We tested crude organic extracts and purified fractions isolated from Ircinia campana, I. felix, and I. strobilina at naturally occurring concentrations in laboratory and field feeding assays to determine their palatability to generalist fish predators. We also used a qualitative technique to test the crude volatile fraction from I. felix and I. strobilina and dimethylsulfide in laboratory feeding assays. Crude organic extracts of all three species deterred feeding of fishes in both aquarium and field experiments. Bioassay-directed fractionation resulted in the isolation of the FTA fraction as the sole active fraction of the nonvolatile crude extract for each species, and further assays of subfractions suggested that feeding deterrent activity is shared by the FTAs. FTAs deterred fish feeding in aquarium assays at concentrations as low as 0.5 mg/ml (fraction B, variabilin), while the natural concentrations of combined FTA fractions were >5.0 mg/ml for all three species. In contrast, natural mixtures of volatiles transferred from sponge tissue to food pellets and pure dimethylsulfide incorporated into food pellets were readily eaten by fish in aquarium assays. Although FTAs may play other ecological roles in Ircinia spp., these compounds are effective as defenses against potential predatory fishes. Volatile compounds may serve other defensive functions (e.g., antimicrobial, antifouling) but do not appear to provide a defense against fish predators.

Phylogenetic relationships among zooxanthellae (Symbiodinium) associated to excavating sponges (Cliona spp.) reveal an unexpected lineage in the Caribbean

Phylogenetic relationships of symbiotic dinoflagellate lineages, distributed in all tropical and subtropical seas, suggest strategies for long distance dispersal but at the same time strong host specialization. Zooxanthellae (Symbiodinium: Dinophyta), which are associated to diverse shallow-water cnidarians, also engage in symbioses with some sponge species of the genus Cliona. In the Caribbean, zooxanthellae-bearing Cliona has recently become abundant due to global warming, overfishing, and algae abundance. Using molecular techniques, the symbionts from five excavating species (Clionacaribbaea, C. tenuis, C. varians, C. aprica and C. laticavicola) from the southern and southwestern Caribbean were surveyed. Several DNA sequence regions were used in order to confirm zooxanthellae identity; 18S rDNA, domain V of chloroplast large subunit (cp23S), internal transcribed spacer 2 (ITS2), and ITS2 secondary structure. Sequence analyses corroborated the presence of three zooxanthellae clades: A, B, and G. Presence of clades A and B in common boring sponges of the Caribbean fit with the general pattern of the province. The discovery of clade G for the first time in any organism of the Atlantic Ocean leads us to consider this unusual finding as a phylogenetic relict through common ancestors of sponge clades or an invasion of the sponge from the Indo-Pacific.

Exudation of low molecular weight compounds (thiobismethane, methyl isocyanide, and methyl isothiocyanate) as a possible chemical defense mechanism in the marine sponge Ircinia felix

The volatile constituents of the marine sponge Ircinia felix were obtained by dynamic headspace extraction and analyzed by HRGC, HRGC-MS and HRGC-Odor at sniffing port. Fifty-nine volatiles were identified for the first time in the odor of this sponge. Hydrocarbons (32.9%), alcohols (17.8%) and carbonyl compounds (16.0%) predominated in the sponge volatile profile, followed by esters (11.6%), halogen compounds (8.6%), ethers (7.7%), nitrogen and/or sulfur compounds (4.6%) and carboxylic acids (0.8%). Among the identified volatiles, thiobismethane (commonly known as dimethylsulfide), methyl isocyanide and methyl isothiocyanate were found to be responsible for the nauseating and toxic smell emitted by the sponge and for the antimicrobial activity detected in the volatile extract. Exudation experiments in aquarium and in situ conditions revealed that thiobismethane, methyl isocyanide and methyl isothiocyanate are continuously released by the sponge. Upon injury, the concentration of these volatiles increased strongly. Hence, these substances form a chemical protective barrier which may help these sponges avoid fouling, compete for space, prevent infection in the short term, and/or signal generalist predators regarding the existence of other toxic substances in the internal tissues.

Phylogenetic analysis of the order Halichondrida (Porifera, Demospongiae), using 3β-hydroxysterols as chemical characters

To analyze and extract phylogenetic information from chemical characters, the 3b-hydroxysterol composition of 12 sponge species of various families of the Poriferan order Halichondrida (sensu lato), was determined. The current definition of the order is ambiguous, and its classification remains in a state of flux and hence, further chemical characters may aid in stabilizing it. Axinella corrugata, Pseudaxinella reticulata, P. explicata, Ptilocaulis walpersi, Dragmaxia undata (family Axinellidae), Myrmekioderma rea, M. gyroderma(family Desmoxyidae), Scopalina ruetzleri (family Dictyonellidae), Halichondria lutea, H. magniconulosa, Topsentia ophiraphidites and Petromica ciocalyptoides (family Halichondriidae) were collected in the Santa Marta area, Colombian Caribbean. Cladistic principles were used to advance a hypothesis of genealogical relationship among the studied species and to contrast current classifications at various taxonomic ranks. Sterol identity and various sterol properties (number of carbons, nuclei type, saturations and substitutions of the lateral chain) were coded as discrete characters according to their relative abundance in 4% ranges, for each of the studied species, and for an out group of 41 species of nine orders whose sterol fractions had been published. Of more than 300 codified characters, 74 constituted unique evolutionary novelties within the studied in group. Fifty-one of these characters (including 15 sterols) uniquely defined six species. Twelve sets of novel characters were shared exclusively by two or more species of the studied group and were thus informative in a phylogenetic sense. The cladogram, built by hand, had seven characters that were compatible with each other and were shared by two or more species (i.e. synapomorphies) and five that were incompatible with two or more. However

How a collaborative integrated taxonomic effort has trained new spongiologists and improved knowledge of Martinique Island (French Antilles, eastern Caribbean Sea) marine biodiversity

Although sponges are important components of benthic ecosystems of the Caribbean Sea, their diversity remained poorly investigated in the Lesser Antilles. By organizing a training course in Martinique, we wanted both to promote taxonomy and to provide a first inventory of the sponge diversity on this island. The course was like a naturalist expedition, with a field laboratory and a classroom nearby. Early-career scientists and environmental managers were trained in sponge taxonomy. We gathered unpublished data and conducted an inventory at 13 coastal sites. We explored only shallow water habitats (0–30 m), such as mangroves, reefs or rocky bottoms and underwater caves. According to this study, the sponge fauna of Martinique is currently represented by a minimum of 191 species, 134 of which we could assign species names. One third of the remaining non-identified sponge species we consider to be new to science. Martinique appears very remarkable because of its littoral marine fauna harboring sponge aggregations with high biomass and species diversity dominating over coral species. In mangroves, sponges cover about 10% of the surface of subtidal roots. Several submarine caves are true reservoirs of hidden and insufficiently described sponge diversity. Thanks to this new collaborative effort, the Eastern Caribbean has gained a significant increase of knowledge, with sponge diversity of this area potentially representing 40% of the total in the Caribbean Sea. We thus demonstrated the importance of developing exploratory and educational research in areas historically devoid of biodiversity inventories and systematics studies. Finally, we believe in the necessity to consider not only the number of species but their distribution in space to evaluate their putative contribution to ecosystem services and our willingness to preserve them.

Validation and evaluation of an HPLC methodology for the quantification of the potent antimitotic compound (+)‐discodermolide in the Caribbean marine sponge Discodermia dissoluta

The sponge Discodermia dissoluta is the source of the potent antimitotic compound (+)-discodermolide. The relatively abundant and shallow populations of this sponge in Santa Marta, Colombia, allow for studies to evaluate the natural and biotechnological supply options of (+)-discodermolide. In this work, an RP-HPLC-UV methodology for the quantification of (+)-discodermolide from sponge samples was tested and validated. Our protocol for extracting this compound from the sponge included lyophilization, exhaustive methanol extraction, partitioning using water and dichloromethane, purification of the organic fraction in RP-18 cartridges and then finally retrieving the (+)-discodermolide in the methanol-water (80:20 v/v) fraction. This fraction was injected into an HPLC system with an Xterra RP-18 column and a detection wavelength of 235 nm. The calibration curve was linear, making it possible to calculate the LODs and quantification in these experiments. The intra-day and inter-day precision showed relative standard deviations lower than 5%. The accuracy, determined as the percentage recovery, was 99.4%. Nine samples of the sponge from the Bahamas, Bonaire, Curaçao and Santa Marta had concentrations of (+)-discodermolide ranging from 5.3 to 29.3 microg/g(-1) of wet sponge. This methodology is quick and simple, allowing for the quantification in sponges from natural environments, in situ cultures or dissociated cells.

MAIN STEROLS FROM THE OPHIUROIDS OPHIOCOMA ECHINATA, OPHIOCOMA WENDTII, OPHIOPLOCUS JANUARII AND OPHIONOTUS VICTORIAE

Abstract Sterol compositions of the cold water ophiuroids Ophioplocus januarii and Ophionotus victoriae and of the tropical ophiuroids Ophiocoma echinata and Ophiocoma wendtii are reported. The four sterol mixtures contain Δ 5 mono- and di-unsaturated common 3β-hydroxy-sterols. Ophioplocus januarii and O. victoriae contain 24-methylcholesta-5,24(28)-dien-3β-ol and 24ζ-ethylcholesta-5,24(28)-dien-3β-ol in higher abundance than in O. echinata . These sterols were not found in O. wendtii . An interesting finding is the presence of Δ 5,24(28) -24- n -propylidenecholesterol in 7.6% in Ophionotus victoriae .

Cliona acephala (Porifera: Demospongiae: Clionaida), a new encrusting excavating reef sponge from the Colombian Caribbean belonging to the Cliona viridis species complex

Several groups of sponges are able to excavate galleries and tunnels in calcareous substrata such as limestone rock, shells, calcareous algae and coral skeletons. Within the genus Cliona, some species share the common traits of being brown to olive-green in color, and harboring photosynthetic, unicellular dinoflagellates (zooxanthellae). These Cliona spp. have been grouped as the Cliona viridis species complex. Several species of this complex completely encrust the excavated substratum with a thin veneer of tissue and, when colonizing dead exposed parts of live coral colonies, they are able to undermine or overgrow and thus kill live coral tissue as they advance predominantly laterally. In the course of our taxonomic and ecological studies of Caribbean brown to brown-black encrusting Cliona, we found an as yet undescribed species that stands out by having tylostyle megasclere spicules with narrow heads and lacking the usual microsclere spicule complement of spirasters. This species, named and described here Cliona acephala n. sp., has so far been found exclusively in the Santa Marta area, Caribbean coast of Colombia. Previous studies with ITS2 ribosomal DNA showed it to be genetically distinct from other Caribbean encrusting species belonging to the Cliona viridis species complex, vis. Cliona aprica, Cliona caribbaea, Cliona tenuis and Cliona varians, but making it genetically closer to Indo-Pacific Cliona orientalis. An intriguing possibility, to be addressed with further studies, is that C. acephala n. sp. may have been introduced to the Caribbean. However, until proved otherwise, we regard the material presently described as distinct.

Unraveling the structure and composition of Varadero Reef, an improbable and imperiled coral reef in the Colombian Caribbean

Coral reefs are commonly associated with oligotrophic, well-illuminated waters. In 2013, a healthy coral reef was discovered in one of the least expected places within the Colombian Caribbean: at the entrance of Cartagena Bay, a highly-polluted system that receives industrial and sewage waste, as well as high sediment and freshwater loads from an outlet of the Magdalena River (the longest and most populated river basin in Colombia). Here we provide the first characterization of Varadero Reef’s geomorphology and biological diversity. We also compare these characteristics with those of a nearby reference reef, Barú Reef, located in an area much less influenced by the described polluted system. Below the murky waters, we found high coral cover of 45.1% (±3.9; up to 80% in some sectors), high species diversity, including 42 species of scleractinian coral, 38 of sponge, three of lobster, and eight of sea urchin; a fish community composed of 61 species belonging to 24 families, and the typical zonation of a Caribbean fringing reef. All attributes found correspond to a reef that, according to current standards should be considered in “good condition”. Current plans to dredge part of Varadero threaten the survival of this reef. There is, therefore, an urgent need to describe the location and characteristics of Varadero as a first step towards gaining acknowledgement of its existence and garnering inherent legal and environmental protections.

Taxonomy of the Caribbean sponge Dragmacidon reticulatum (Ridley & Dendy, 1886) (Porifera, Demospongiae, Axinellida), with the description of a new species.

Although there is a long history of taxonomic investigation in Caribbean sponges, there are still many undescribed species. Furthermore, field observations and corroborating morphological analyses are revealing that what was believed to be single, somewhat variable species, may consist of two or more species, often easier to distinguish once well characterized. This is the case for Dragmacidon reticulatum (Ridley & Dendy, 1886) (Porifera, Demospongiae, Axinellida, Axinellidae), a rather well-known sponge, with an ample distribution and presence in rocky and reef environments of the tropical and subtropical Western Atlantic, with local records in the majority of the countries of the area, from Bermuda to Brazil. Field observations and a detailed review of material from different areas, including some type specimens, led us to the distinction of two different species in terms of external morphology, size of spicules, and skeletal architecture. The distinction was confirmed in the Bahamas and Santa Marta, Colombia, where the two species coexist. One of the species is Dragmacidon reticulatum sensu stricto, but for the other there is need to erect a new name, for which we propose Dragmacidon alvarezae n. sp. The purpose of the present work is to describe, illustrate and compare these two species.