Vertical Transmission of a Phylogenetically Complex Microbial Consortium in the Viviparous Sponge \u3cem\u3eIrcinia Felix\u3c/em\u3e

Many marine demosponges contain large amounts of phylogenetically complex yet highly sponge-specific microbial consortia within the mesohyl matrix, but little is known about how these microorganisms are acquired by their hosts. Settlement experiments were performed with the viviparous Caribbean demosponge Ircinia felix to investigate the role of larvae in the vertical transmission of the sponge-associated microbial community. Inspections by electron microscopy revealed large amounts of morphologically diverse microorganisms in the center of I. felix larvae, while the outer rim appeared to be devoid of microorganisms. In juveniles, microorganisms were found between densely packed sponge cells. Denaturing gradient gel electrophoresis (DGGE) was performed to compare the bacterial community profiles of adults, larvae, and juvenile sponges. Adults and larvae were highly similar in DGGE band numbers and banding patterns. Larvae released by the same adult individual contained highly similar DGGE banding patterns, whereas larvae released by different adult individuals showed slightly different DGGE banding patterns. Over 200 bands were excised, sequenced, and phylogenetically analyzed. The bacterial diversity of adult I. felix and its larvae was comparably high, while juveniles showed reduced diversity. In total, 13 vertically transmitted sequence clusters, hereafter termed “IF clusters,” that contained sequences from both the adult sponge and offspring (larvae and/or juveniles) were found. The IF clusters belonged to at least four different eubacterial phyla and one possibly novel eubacterial lineage. In summary, it could be shown that in I. felix, vertical transmission of microorganisms through the larvae is an important mechanism for the establishment of the sponge-microbe association

Episymbiotic microbes as food and defence for marine isopods : unique symbioses in a hostile environment

Author Posting. © Royal Society, 2005. This article is posted here by permission of Royal Society for personal use, not for redistribution. The definitive version was published in Proceedings of the Royal Society of London B 272 (2005): 1209-1216, doi:10.1098/rspb.2005.3082.Symbioses profoundly affect the diversity of life, often through novel biochemical services that symbionts provide to their hosts. These biochemical services are typically nutritional enhancements and less commonly defensive, but rarely both simultaneously. On the coral reefs of Papua New Guinea, we discovered unique associations between marine isopod crustaceans (Santia spp.) and episymbiotic microbes. Transmission electron microscopy and pigment analyses show that episymbiont biomass is dominated by large (20–30μm) cyanobacterial cells. The isopods consume these photosymbionts and ‘cultivate’ them by inhabiting exposed sunlit substrates, a behaviour made possible by symbionts' production of a chemical defence that is repulsive to fishes. Molecular phylogenetic analyses demonstrated that the symbiotic microbial communities are diverse and probably dominated in terms of population size by bacteria and small unicellular Synechococcus-type cyanobacteria. Although largely unknown in the oceans, defensive symbioses probably promote marine biodiversity by allowing niche expansions into otherwise hostile environments.This work was supported by an NSF Predoctoral Fellowship awarded to J.B.W and a grant from the North Carolina Biotechnology Center to N.L and Richard Manderville (Wake Forest University)

Palatability and Chemical Defense of Marine Invertebrate Larvae

Risk of larval mortality is a critical component of models and debates concerning the ecology and evolution of the differing reproductive characteristics exhibited by marine invertebrates. In these discussions, predation often is assumed to be a major source of larval mortality. Despite limited empirical support, most marine larvae are thought to be palatable and broadly susceptible to generalist predators. Previous studies of larval-planktivore interactions have focused primarily on larvae that typically feed, grow, and develop for weeks to months in the plankton. Such planktotrophic species commonly produce large numbers of small larvae that disperse over vast distances. In contrast, the nonfeeding lecithotrophic larvae from sessile invertebrates that brood are often large and conspicuous, lack morphological defenses, and have limited dispersal distances because they typically are competent to settle minutes to hours after spawning. Interactions between lecithotrophic larvae and consumers are not well studied. This has limited the ability of previous authors to test broad generalities about marine larvae. We show that brooded larvae of Caribbean sponges (11 species) and gorgonians (three species) as well as brooded larvae of temperate hydroids (two species) and a bryozoan are unpalatable to co-occurring fishes. In contrast, brooded larvae of temperate ascidians (three species), a temperate sponge, and Caribbean hard corals (three species) are readily consumed by fishes, as are larvae from four of six species of synchronous broadcast-spawning gorgonians from the Florida Keys. Frequencies of survivorship for larvae attacked and rejected by fishes were high and statistically indistinguishable from frequencies for unattacked control larvae. Frequency of metamorphosis (when it occurred) of rejected larvae never differed significantly from that of unattacked control larvae. Assays testing for larval vs. adult chemical defenses for five species with distasteful larvae showed that larvae of all five species were chemically distasteful to fishes, whereas only three of five adult extracts deterred fish feeding. A comparison of larval palatability among chemically rich taxa showed that brooded larvae were significantly more likely to be unpalatable (86% of the species tested) than larvae of broadcasters (33%), and that palatable larvae were rarely released during the day (23%) while unpalatable larvae usually were (89%). Additionally, the frequency of bright coloration was high (60%) for unpalatable larvae and low (0%) for palatable larvae, suggesting that unpalatable larvae often may be aposematically colored. Results of this broad survey cast doubt on the widely accepted notion that virtually all marine larvae are suitable prey for most generalized planktivores. Among species that do not chemically or physically protect larvae against fishes, selection appears to favor the release of larvae at night, or the production of smaller more numerous offspring that grow and develop at sea as a way of escaping consumer-rich benthic habitats. Because distasteful larvae are not similarly constrained, distasteful species should exhibit reproductive and larval characteristics selected more by the fitness-related consequences of larval development mode and dispersal distance than by the necessity of avoiding benthic predators

Chemical defense among hydroids on pelagic Sargassum:predator deterrence and absorption of solar UV radiation by secondary metabolites

The pelagic Sargassum community of the western Atlantic Ocean is species rich, with high densities of predatory fishes and invertebrates living in close association with the floating seaweed. Little, however, is known about predator-prey interactions among Sargassum inhabitants and the factors that might help maintain this species richness. To assess how predators may affect the abundance of sessile Sargassum epiphytes, and how these epiphytes may defend themselves against predators, we examined interactions between the most abundant small predator associated with Sargassum mats, the filefish Monacanthus hispidus, and 4 epiphytic hydroid species. This fish was the only Sargassum-associated predator to significantly consume hydroids in initial assays. When filefish were given a simultaneous choice of all 4 hydroid species, they consumed 40 to 45% of 3 species (Clytia noliformis, Aglaophenia latecarinata, and Tridentata turbinata), but consumed less than 5% of the fourth species, Tridentata marginata. Filefish consistently rejected small portions of T. marginata colonies, but consumed a palatable control food. Bioassay-guided fractionation of T. marginata extract resulted in the isolation of a single deterrent secondary metabolite, tridentatol A. Three additional metabolites (tridentatols B to D) had no effect on filefish feeding. In addition to the defensive role of tridentatol A, the tridentatols (A to D) strongly absorb damaging solar ultraviolet (UV) radiation, and thus may function as sunscreens. To our knowledge, this is the first demonstration of chemical defenses among the pelagic Sargassum fauna, and the first report that a hydroid secondary metabolite deters consumers. Prey chemical defenses are an important factor in maintaining species richness in many predator-rich communities, but despite being chemically defended from predators, T. marginata was far less abundant than any of the other, more palatable, hydroids. The factors that allow high-preference hydroids to persist in such a predator-rich community remain unknown

Can small rare prey be chemically defended? the case for marine larvae

DOI: 10.2307/1940941©1995 Ecological Society of AmericaMarine larvae are consumed by a wide variety of generalist fishes and particle—feeding invertebrates, but larvae of any particular species probably constitute a small and variable of the diet for these consumers. Because virtually all consumers can ingest small quantities of noxious compounds with minimal detrimental effects, it is uncertain that toxic chemicals in larvae could be consumed in quantities sufficient to select for predator recognition and avoidance. Despite this, chemically defended larvae do occur. We show that, at low doses, secondary metabolites (the didemnins) from adults and larvae of the Caribbean tunicate Trididemnum solidum induced vomiting in fish, resulting in rapid learned aversion of the didemnin—defended food. The particle—feeding anemone Aiptasia pallida did not learn to avoid the chemically defended food. When anemones ingested the chemical equivalent of 15 larvae/d, representing <2% of the mass of their total daily diet, the didemnins in the "larvae" significantly reduced: (1) growth of adults by 82%, (2) combined growth of adults and daughter clones by 76%, (3) production of daughter clones by 44%, and (4) average mass of individual daughter clones by 41%. At higher water temperatures, anemones cloned more rapidly, but the negative effects of consuming didemnins also occurred more rapidly. Significant differences in the number of daughter clones produced between treatment and control anemones occurred after only 4 d at seawater temperatures of 27°—29°C vs. 32 d at seawater temperatures of 18°—21°C. Thus consumption of even very small quantities of secondary metabolites can decrease consumer fitness substantially and select for predators that recognize and avoid chemically defended larvae, as do many consumers that co—occur with Trididemnum solidum larvae. This is the first rigorous demonstration that consumption of marine secondary metabolites can decrease consumer fitness when ingested at ecologically realistic doses

Potent cytotoxins produced by a microbial symbiont protect host larvae from predation

Larvae of the sessile marine invertebrate Bugula neritina (Bryozoa) are protected by an effective chemical defense. From the larvae, we isolated three bryostatin-class macrocyclic polyketides, including the novel bryostatin 20, that deterred feeding by a common planktivorous fish that co-occurs with B. neritina . A unique bacterial symbiont of B. neritina , Endobugula sertula , was hypothesized as the putative source of the bryostatins. We show that: (1) bryostatins are concentrated in B. neritina larvae and protect them against predation by fish; (2) the adults are not defended by bryostatins; and (3) E. sertula produces bryostatins. This study represents the first example from the marine environment of a microbial symbiont producing an anti-predator defense for its host and, in this case, specifically for the host’s larval stage, which is exceptionally vulnerable to predators.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47703/1/442_2004_Article_1487.pd

Constraints on chemically-mediated coevolution: multiple functions for seaweed secondary metabolites.

DOI: 10.2307/1940635©1995 Ecological Society of AmericaDiterpene alcohols produced by the brown seaweed Dictyota menstrualis deter feeding by numerous species of abundant herbivores. Here we show that these same compound also may prevent fouling organisms from colonizing the surface of this alga. In the field, Dictyota menstrualis plants were less frequently and less heavily fouled than any of the other common seaweed species investigated. In laboratory assays, larvae of the common fouling bryozoan Bugula neritina failed to settle on Dictyota even though they contacted its surface as often as they contacted the surface of a preferred host alga. Rejection occurred only after direct contact with the alga's surface. Rejection of Dictyota was not mediated by water—borne chemical cues or by surface wettability (a physical property of the surface that can affect fouling). The lipid—soluble extract from surface rubbings of Dictyota inhibited larval settlement when placed on other surfaces and contained the diterpene alcohols pachydictyol A and dictyol E. Larvae exposed to these compounds experienced mortality, abnormal development, or reduced rates of development. Although the potential for chemically mediated coevolution between plants and herbivores has been the focus of scores of previous investigations, such coevolution will depend on selection altering the chemical defenses of the plant following the evolution of resistance by herbivores. Such a reciprocal response will be constrained if compounds play multiple roles that are ecologically important. Dictyota produces secondary metabolites that are broadly defensive against a wide variety of consumers and fouling organisms. Although certain consumers may evolve resistance to these metabolites, it is unclear that feeding by these consumers will result in reciprocal responses from the plant. We suggest that coevolved interactions may be uncommon, and that many interactions that appear to be coevolved may result from fortuitous and opportunistic preadaptations

The pathology of sponge orange band disease affecting the Caribbean barrel sponge Xestospongia muta

The aim of this study was to examine sponge orange band (SOB) disease affecting the prominent Caribbean sponge Xestospongia muta. Scanning and transmission electron microscopy revealed that SOB is accompanied by the massive destruction of the pinacoderm. Chlorophyll a content and the main secondary metabolites, tetrahydrofurans, characteristic of X. muta, were significantly lower in bleached than in healthy tissues. Denaturing gradient gel electrophoresis using cyanobacteria-specific 16S rRNA gene primers revealed a distinct shift from the Synechococcus/Prochlorococcus clade of sponge symbionts towards several clades of unspecific cyanobacteria, including lineages associated with coral disease (i.e. Leptolyngbya sp.). Underwater infection experiments were conducted by transplanting bleached cores into healthy individuals, but revealed no signs of SOB development. This study provided no evidence for the involvement of a specific microbial pathogen as an etiologic agent of disease; hence, the cause of SOB disease in X. muta remains unidentified

The HMA-LMA dichotomy revisited: an electronmicroscopical survey of 56 sponge species

The dichotomy between high microbial abundance (HMA) and low microbial abundance (LMA) sponges has been long recognized. In the present study, 56 sponge species from three geographic regions (greater Caribbean, Mediterranean, Red Sea) were investigated by transmission electron microscopy for the presence of microorganisms in the mesohyl matrix. Additionally, bacterial enumeration by DAPI-counting was performed on a subset of samples. Of the 56 species investigated, 28 were identified as belonging to the HMA and 28 to the LMA category. The sponge orders Agelasida and Verongida consisted exclusively of HMA species, and the Poecilosclerida were composed only of LMA sponges. Other taxa contained both types of microbial associations (e.g., marine Haplosclerida, Homoscleromorpha, Dictyoceratida), and a clear phylogenetic pattern could not be identified. For a few sponge species, an intermediate microbial load was determined, and the microscopy data did not suffice to reliably determine HMA or LMA status. To experimentally determine the HMA or LMA status of a sponge species, we therefore recommend a combination of transmission electron microscopy and 16S rRNA gene sequence data. This study significantly expands previous reports on microbial abundances in sponge tissues and contributes to a better understanding of the HMA-LMA dichotomy in sponge-microbe symbioses