Molecular evidence of digestion and absorption of epibiotic bacterial community by deep-sea crab Shinkaia crosnieri

The hydrothermal vent crab Shinkaia crosnieri is considered to obtain nutrition from the epibiotic bacteria found on the setae, but previous studies have not shown how nutrients can be transferred from the epibionts to the host. In this study, microscopic observations of S. crosnieri intestinal components detected autofluorescent setae fragments and pigmentation derived from the digestion of epibionts in a dye-stained epibiont tracer experiment. An in vitro digestion experiment with epibiotic populations using an intestinal extract demonstrated the degradation of epibiotic cells by digestive enzymes. A phylogenetic analysis showed that many of the bacterial 16S ribosomal RNA gene sequences obtained from the intestine were closely related to the sequences of the epibionts, thus they were probably derived from the epibionts. A stable isotope tracer experiment also indicated that ¹³C assimilated by the epibionts provided a carbon (nutrition) source for the host. Both activity measurements and isotope studies showed that chemosynthetic metabolism by the gut microbial components were inactive. Together with the feeding behaviour of living S. crosnieri, these results indicate that S. crosnieri ingests the epibionts using maxillipeds and assimilates them via its digestive organs as a nutrient source. The results of this study elucidate the mechanism of nutritional transfer in ectosymbiosis between chemosynthetic bacteria and deep-sea invertebrates.     

Epibiotic protozoan communities on juvenile southern king crabs (<Emphasis Type="Italic">Lithodes santolla</Emphasis>) from subantarctic areas

In this study, we describe for the first time the composition of epibiotic protozoan communities on juvenile southern king crabs Lithodes santolla. Basibionts were collected in subantarctic bays near Santa Ana Point (Magellan Strait, Chile). Seven epibiotic protozoan ciliates were found: Ephelota gemmipara, Ephelota gigantea, Podophrya fixa, Acineta tuberosa, Zoothamnium duplicatum, Chilodochona quennerstedti and Gymnodinioides sp. The mean number of epibionts per crab was 99 (maximum 897). Both Ephelota species were the most abundant and most widely distributed epibionts on the crab examined. The lengths of basibionts were correlated with the number of epibionts, indicating a relationship between the age of the crab and the state of epibiosis. Epibionts differed in their distribution patterns along the anteroposterior axis of the crab. Less abundant species tended to occupy sites that are not colonized by the predominant Ephelota species. Maxillipeds and pereiopods were most densely colonized by epibionts. The behaviours of epibionts and basibionts related to epibiosis are discussed. The recent studies about invasive lithodids in Antarctic areas enhance the interest for the epibiotic communities colonizing these crustaceans, which can illustrate changes in biodiversity and state of these environments.     

Hydrozoan and protozoan epibionts on two decapod species, Liocarcinus depurator (Linnaeus, 1758) and Pilumnus hirtellus (Linnaeus, 1761), from Scotland

Crabs of the species Liocarcinus depurator and Pilumnus hirtellus, collected on the west coast of Scotland, showed hydrozoan and protozoan epibionts. The epibionts of Liocarcinus depurator were the protozoan Ephelota plana and the hydrozoans Clytia gracilis and Leuckartiara sp. The epibionts of Pilumnus hirtellus were the protozoans E. plana and Zoothamnium sp. For Liocarcinus depurator the number of Ephelota per crab fluctuated between 0 and 47 and the greatest number of suctorians were located on the chelipeds, carapace and anterior pereiopods. The hydrozoans, for Liocarcinus depurator, showed densities of 0-20 (Clytia gracilis on the second pereiopod) and 0-507 individuals per crab (Leuckartiara sp., principally on chelipeds, carapace and the fourth right pereiopod). For Pilumnus hirtellus, Ephelota plana showed densities between 3 and 56 individuals per crab, the greatest number of suctorians being located on the same areas as on Liocarcinus depurator. There was a density of 10-69 individuals per crab of Zoothamnium sp. on Pilumnus hirtellus (located on the carapace). Ephelota plana has not been observed previously as an epibiont on crustacea, nor had Zoothamnium sp. as an epibiont on Pilumnus hirtellus. Both hydrozoans, Leukartiara and Clytia, have not been previously described as epibionts on Liocarcinus depurator. Data about the morphological characteristics and distribution of these epibionts are included.     

Taxonomy and distribution of the hydrozoan and protozoan epibionts on Pagurus bernhardus (Linnaeus, 1758) (Crustacea, Decapoda) from Scotland

Protozoan and hydrozoan epibionts on the hermit crab Pagurus bernhardus and its shell, collected near Cumbrae Island (Scotland) were studied. The epibionts found were the following: (1) protozoans: the suctorian ciliates Ephelota plana, Acineta compressa, Conchacineta constricta, Corynophrya anisostyla; the peritrich ciliates Cothurnia mobiusi and Zoothamnium plumula; the chonotrich ciliate Chilodochona quennerstedti; and (2) hydrozoans: the species Leuckartiara sp. and Clytia sp. The morphological characteristics of the epibionts were analysed, as well as their taxonomic position. The distribution of epibionts on the crab surface and its shell was studied, and the density and biomass of epibionts were calculated on each anatomic unit. There was a differential distribution according to the type of epibiont: hydrozoans dominated in biovolume and were present mainly on the shell, meanwhile protozoans represented the highest fraction of density and they were found exclusively on the crab (principally on eyes, antennulae, antennae, maxillipeds, pereopods and uropods). The anterior area of the cephalothorax was the most colonized. On this area, the maxillipeds and second pereopods showed the highest densities. The location of each epibiont species was described. There was a correlation between the length of the crab and the total number of hydrozoans. There was a significant correlation between the right and the left units of the crab, taking into consideration the mean densities of epibionts on each anatomical unit. The shell was colonized by the same species of hydrozoa that appeared on the crab, although in a much higher density (mean 3024.38 per shell; 6.9 per crab). There was a significant difference between both species of hydrozoan epibionts with respect to the mean densities on the different areas of the shell. The zone of the shell more occupied by Clytia sp. was the apical zone of the shell, while the highest densities of Leuckartiara sp. were registered near the aperture of the shell. The hydrozoan and protozoan epibiont species found on P. bernhardus in this study represent the first mention of their presence on this hermit crab.     

Effects of epibiosis on consumer–prey interactions

In many benthic communities predators play a crucialrole in the population dynamics of their prey. Surfacecharacteristics of the prey are important forrecognition and handling by the predator. Because theestablishment of an epibiotic assemblage on thesurface of a basibiont species creates a new interfacebetween the epibiotized organism and its environment,we hypothesised that epibiosis should have an impacton consumer–prey interactions. In separateinvestigations, we assessed how epibionts onmacroalgae affected the susceptibility of the latterto herbivory by the urchin Arbacia punctulataand how epibionts on the blue mussel Mytilusedulis affected its susceptibility to predation bythe shore crab Carcinus maenas.Some epibionts strongly affected consumer feedingbehavior. When epibionts were more attractive thantheir host, consumer pressure increased. Whenepibionts were less attractive than their host or whenthey were repellent, consumer pressure decreased. Insystems that are controlled from the top-down,epibiosis can strongly influence community dynamics.For the Carcinus/Mytilus system that westudied, the in situ distribution of epibiontson mussels reflected the epibiosis-determinedpreferences of the predator. Both direct and indirecteffects are involved in determining theseepibiont-prey–consumer interactions.     

Optimal foraging versus shared doom effects: interactive influence of mussel size and epibiosis on predator preference

In the western Baltic Sea, the highly competitive blue mussel Mytilus edulis tends to monopolize shallow water hard substrata. In many habitats, mussel dominance is mainly controlled by the generalist predator Carcinus maenas. These predator-prey interactions seem to be affected by mussel size (relative to crab size) and mussel epibionts.There is a clear relationship between prey size and predator size as suggested by the optimal foraging theory: Each crab size class preferentially preys on a certain mussel size class. Preferred prey size increases with crab size.Epibionts on Mytilus, however, influence this simple pattern of feeding preferences by crabs. When offered similarly sized mussels, crabs prefer Balanus-fouled mussels over clean mussels. There is, however, a hierarchy of factors: the influence of attractive epibiotic barnacles is weaker than the factor ‘mussel size’. Testing small mussels against large mussels, presence or absence of epibiotic barnacles does not significantly alter preferences caused by mussel size. Balanus enhanced crab predation on mussels in two ways: Additional food gain and, probably more important, improvement in handling of the prey. The latter effect is illustrated by the fact that artificial barnacle mimics increased crab predation on mussels to the same extent as do live barnacles.We conclude that crab predation preferences follows the optimal foraging model when prey belong to different size classes, whereas within size classes crab preferences is controlled by epibionts.     

Potential effect of floating dock communities on a South Carolina estuary

A 4-yr study on the motile epibionts colonizing hydroids on a floating dock in a South Carolina estuary indicated that the motile epibiotic community may impact benthic and nektonic community processes. Permanent members of the epibiotic community were preyed upon by fish, especially when the Atlantic silverside (Menidia menidia (L.)) became reproductively active and deposited eggs among the hydroids during the spring. When fish predation reduced the number of amphipods within the epibiotic community, crab megalopa colonized the hydroids. Crabs developed into juveniles and emigrated from the epibiotic community after 6–8 wk residence. The community on floating docks may be an important food source for some nekton and may recruit some benthic invertebrates.Although preferential larval settlement and fish predation are intuitive processes associated with floating dock communities, insufficient research has addressed the relationships in soft-bottom areas devoid of naturally occurring hard substrata.     

Seasonal dynamic of phototrophic epibionts on crustacean zooplankton in a eutrophic reservoir with cyanobacterial bloom

Seasonality of burden and prevalence of phototrophic (microalgal) epibionts Characidiopsis ellipsoidea, Colacium vesiculosum and Colacium sp. on dominating crustacean zooplankton (Daphnia longispina, Cyclops vicinus and Mesocyclops leuckarti) were studied in a small reservoir Bugach with cyanobacterial bloom. The correlations between the seasonal dynamics of prevalence and the dynamics of others biotic and abiotic factors were calculated. The conclusions were as follows. The substrate species, that determined the development of the epibionts on the three studied crustacean zooplankton, was Daphnia longispina (Cladocera). Despite intensive epibiotic infestation of crustacean zooplankton, epibionts did not appear to have caused non-consumptive mortality of the crustacean zooplankton. But they could have contributed to the Daphnia summer decline by increasing mortality due to its consumption by planktivorous fishes. The phototropic epibionts may successfully coexist with cyanobacterial bloom. The possible role of the epibionts in changing nutrient fluxes in pelagic food web is discussed.     

Quorum sensing control of phosphorus acquisition in Trichodesmium consortia

Colonies of the cyanobacterium Trichodesmium are abundant in the oligotrophic ocean, and through their ability to fix both CO₂ and N₂, have pivotal roles in the cycling of carbon and nitrogen in these highly nutrient-depleted environments. Trichodesmium colonies host complex consortia of epibiotic heterotrophic bacteria, and yet, the regulation of nutrient acquisition by these epibionts is poorly understood. We present evidence that epibiotic bacteria in Trichodesmium consortia use quorum sensing (QS) to regulate the activity of alkaline phosphatases (APases), enzymes used by epibionts in the acquisition of phosphate from dissolved-organic phosphorus molecules. A class of QS molecules, acylated homoserine lactones (AHLs), were produced by cultivated epibionts, and adding these AHLs to wild Trichodesmium colonies collected at sea led to a consistent doubling of APase activity. By contrast, amendments of (S)-4,5-dihydroxy-2,3-pentanedione (DPD)—the precursor to the autoinducer-2 (AI-2) family of universal interspecies signaling molecules—led to the attenuation of APase activity. In addition, colonies collected at sea were found by high performance liquid chromatography/mass spectrometry to contain both AHLs and AI-2. Both types of molecules turned over rapidly, an observation we ascribe to quorum quenching. Our results reveal a complex chemical interplay among epibionts using AHLs and AI-2 to control access to phosphate in dissolved-organic phosphorus.     

The symbiotic role of marine microbes on living surfaces

Every surface immersed in the sea rapidly becomes covered with a biofilm. On inanimate surfaces, this is often followed by colonisation by larger organisms, and general macrofouling. On the other hand, the majority of marine organisms remain relatively free from macrofouling, although some may be covered in a thin film of epibiotic bacteria. The role of these bacteria in maintaining the health of the host has received little attention. Here we describe an ecological role for epibiotic bacteria from seaweed surfaces. These epibionts may play a protective role, releasing compounds into the surrounding seawater that help prevent extensive fouling of the surface. These compounds may also have industrial and medical applications. The relative ease of culturing these microbes, compared to other bacteria that produce active compounds suggests seaweed-associated bacteria may be useful in bioprocess applications, such as the production of antimicrobial or antifouling compounds.     

On the evolutionary ecology of symbioses between chemosynthetic bacteria and bivalves

Mutualistic associations between bacteria and eukaryotes occur ubiquitously in nature, forming the basis for key ecological and evolutionary innovations. Some of the most prominent examples of these symbioses are chemosynthetic bacteria and marine invertebrates living in the absence of sunlight at deep-sea hydrothermal vents and in sediments rich in reduced sulfur compounds. Here, chemosynthetic bacteria living in close association with their hosts convert CO₂ or CH₄ into organic compounds and provide the host with necessary nutrients. The dominant macrofauna of hydrothermal vent and cold seep ecosystems all depend on the metabolic activity of chemosynthetic bacteria, which accounts for almost all primary production in these complex ecosystems. Many of these enigmatic mutualistic associations are found within the molluscan class Bivalvia. Currently, chemosynthetic symbioses have been reported from five distinct bivalve families (Lucinidae, Mytilidae, Solemyidae, Thyasiridae, and Vesicomyidae). This brief review aims to provide an overview of the diverse physiological and genetic adaptations of symbiotic chemosynthetic bacteria and their bivalve hosts.     

Adaptations to Hydrothermal Vent Life in Kiwa tyleri,a New Species of Yeti Crab from the East Scotia Ridge,Antarctica

Hydrothermal vents in the Southern Ocean are the physiologically most isolated chemosynthetic environments known. Here, we describe Kiwa tyleri sp. nov., the first species of yeti crab known from the Southern Ocean. Kiwa tyleri belongs to the family Kiwaidae and is the visually dominant macrofauna of two known vent sites situated on the northern and southern segments of the East Scotia Ridge (ESR). The species is known to depend on primary productivity by chemosynthetic bacteria and resides at the warm-eurythermal vent environment for most of its life; its short-range distribution away from vents (few metres) is physiologically constrained by the stable, cold waters of the surrounding Southern Ocean. Kiwa tylerihas been shown to present differential life history adaptations in response to this contrasting thermal environment. Morphological adaptations specific to life in warm-eurythermal waters, as found on – or in close proximity of – vent chimneys, are discussed in comparison with adaptations seen in the other two known members of the family (K. hirsuta, K. puravida), which show a preference for low temperature chemosynthetic environments.     

New genus and species of brachyuran crab from the southern East Pacific Rise (Crustacea Decapoda Brachyura Bythograeidae)

A new genus and species of a brachyuran crab, Allograea tomentosa n. gen. and n. sp., of the family Bythograeidae, collected at a hydrothermal vent locality south of the Easter Microplate (31 degrees 09'S) on the southern East Pacific Rise, is described. The genus Allograea is distinguished by lacking modifications of the fronto-orbital region, and by the absence of coloured fields and setal patches on the carapace and chelipeds. The body and legs are smooth but densely covered by a brown tomentum, the eyestalks are not reduced, but the cornea is unfaceted and unpigmented. Comparison of mitochondrial DNA sequences among all known bythograeid genera confirms the novelty of this taxon.     

Novel epibiotic thiothrix bacterium on a marine amphipod

Comparative analysis of the 16S rRNA gene and fluorescent in situ hybridization (FISH) was used to identify epibiotic filamentous bacteria living on the marine amphipod crustacean Urothoe poseidonis. The epibionts belong to the gamma proteobacteria and represent a novel marine phylotype within the genus Thiothrix. FISH and denaturing gradient gel electrophoresis revealed that the Thiothrix filaments are present on the majority of the amphipods examined.     

Epibiont protozoan communities on Caridina lanceolata (Crustacea, Decapoda) from the Malili lakes of Sulawesi (Indonesia)

The epibiont protozoan communities living on the freshwater shrimp Caridina lanceolata Woltereck, 1937a from the three major lakes (Towuti, Matano and Mahalona) of the Malili lake system (Sulawesi, Indonesia) were studied. The number of epibionts varied between 2 and 971 per shrimp. Seven protozoan ciliate species were found: Acineta sulawesiensis n. sp., Cothurnia sp., Zoothamnium sp. (in all three lakes), Vorticella sp. (Lake Mahalona and Lake Matano), Opercularia sp. (Lake Mahalona), Epistylis sp. (Lake Mahalona and Lake Matano), and Podophrya sp. (Lake Mahalona). Although these ciliates had been found previously on other crustaceans, they have not been observed as epibionts on Caridina H. Milne Edwards, 1837. The distribution of the different epibiont species on the anatomical units of the shrimp was analyzed in each lake. There is a statistical significant difference between the three lakes in respect to the number of epibionts on each anatomical unit of all analyzed shrimps. The total and mean densities of each epibiont species on the different analyzed shrimps showed a significant difference between the three lakes; i.e., the presence of each epibiont species on the population of C. lanceolata varied from one lake to another. In Lake Towuti the highest density of epibionts was found on the anterior part of the shrimp body (rostrum, antennae, antennulae and eyes) (32.41%), while in the other two lakes, the highest colonization corresponded to the maxillipeds (31.56% Lake Matano, 40.89% Lake Mahalona). In Lake Towuti the rest of epibionts colonized mainly maxillipeds and pleopods (both 45.76% of epibionts). In Lake Matano, other epibionts were distributed principally on the anterior part of the body and pleopods (in total 57.18% of epibionts). In Lake Mahalona, other epibionts were divided among the anterior part of the body, pereiopods and pleopods (in total 57.39% of the epibionts). Uropods and telson were the units less colonized in Lake Matano (3.64%) and Lake Mahalona (1.72%), while in Lake Towuti, they presented a moderate density (13.18% of the epibionts). Taking into account the distribution of epibionts along the antero-posterior axis of the shrimp, considering the different anatomical units, there was a significant correlation between the three lakes. This fact indicates that, in the three lakes, the colonization on C. lanceolata followed a similar distribution pattern, independently of the epibiont species present. The comparison between the distributions of the same epibiont species along the longitudinal axis of the shrimp on the diverse lakes showed that they correlated respect to their density values on the anatomical units of the shrimp. Diverse aspects of the colonization patterns are discussed. Morphological features, taxonomic identification, and particular distribution of the epibiont species in each lake are also included.     

Pathways of carbon and energy metabolism of the epibiotic community associated with the deep-sea hydrothermal vent shrimp Rimicaris exoculata

Background

The shrimp Rimicaris exoculata dominates the faunal biomass at many deep-sea hydrothermal vent sites at the Mid-Atlantic Ridge. In its enlarged gill chamber it harbors a specialized epibiotic bacterial community for which a nutritional role has been proposed.

Methodology/Principal Findings

We analyzed specimens from the Snake Pit hydrothermal vent field on the Mid-Atlantic Ridge by complementing a 16S rRNA gene survey with the analysis of genes involved in carbon, sulfur and hydrogen metabolism. In addition to Epsilon- and Gammaproteobacteria, the epibiotic community unexpectedly also consists of Deltaproteobacteria of a single phylotype, closely related to the genus Desulfocapsa. The association of these phylogenetic groups with the shrimp was confirmed by fluorescence in situ hybridization. Based on functional gene analyses, we hypothesize that the Gamma- and Epsilonproteobacteria are capable of autotrophic growth by oxidizing reduced sulfur compounds, and that the Deltaproteobacteria are also involved in sulfur metabolism. In addition, the detection of proteobacterial hydrogenases indicates the potential for hydrogen oxidation in these communities. Interestingly, the frequency of these phylotypes in 16S rRNA gene clone libraries from the mouthparts differ from that of the inner lining of the gill chamber, indicating potential functional compartmentalization.

Conclusions

Our data show the specific association of autotrophic bacteria with Rimicaris exoculata from the Snake Pit hydrothermal vent field, and suggest that autotrophic carbon fixation is contributing to the productivity of the epibiotic community with the reductive tricarboxylic acid cycle as one important carbon fixation pathway. This has not been considered in previous studies of carbon fixation and stable carbon isotope composition of the shrimp and its epibionts. Furthermore, the co-occurrence of sulfur-oxidizing and sulfur-reducing epibionts raises the possibility that both may be involved in the syntrophic exchange of sulfur compounds, which could increase the overall efficiency of this epibiotic community.     

Epibiotic bacteria associated with the recently discovered Yeti crab, Kiwa hirsuta

The Yeti crab, Kiwa hirsuta Macpherson et al., is the single known species in a recently discovered crab family Kiwaidae (Decapoda: Galatheoidea) from deep-sea hydrothermal vents. Its chelipeds, walking legs and the ventral surface of its cephalothorax are covered with dense setae that, in turn, are covered with clusters of filamentous bacteria, making the crab appear extraordinarily 'hairy'. Electron microscopy revealed dense bacterial clusters attached to the chitinous outer layer of the setae. Molecular phylogenetic analyses revealed the setae-associated bacteria to be dominated by epsilon-Proteobacteria ( approximately 56% of the recovered ribotypes), gamma-Proteobacteria ( approximately 25%) and Bacteroidetes ( approximately 10%). Fluorescence in situ microscopy confirmed the attachment of filamentous epsilon-Proteobacteria on setae, but no specialized morphological structures appeared to exist for bacterial attachment. Key enzymes involved in the reductive tricarboxylic acid cycle (ATP-dependent citrate lyase) and sulfite oxidation or dissimilatory sulfate reduction (bidirectional APS reductase) were detected. Consequently, the potential for carbon fixation and cycling of reduced and oxidized sulfur appear to exist in the dense microflora that grows on the crab's setae.     

Epibiotic macroalgae on the scallop <Emphasis Type="Italic">Mizuhopecten yessoensis</Emphasis> in Peter the Great Bay,Sea of Japan

The composition and structure of the epibiotic flora of the Japanese scallop Mizuhopecten yessoensis were studied on the basis of data from long-term (1979 to 2007) observations on the scallop beds in Peter the Great Bay (Sea of Japan). In all, 52 species of macroalgae belonging to three phyla were found on the scallop shells; 3 species were new records for the benthic flora of the area studied. Red algae constituted the bulk of the species richness of algal epibionts; brown algae were represented by the lowest number of species. Species of Chlorophyta predominated in terms of biomass; species of Rhodophyta were found in lower numbers. The main form of the thallus of epibiotic algae was bushy or filamentous. The ratio of common to rare species was 30 : 70. As compared to the benthic flora, the epibiotic flora on the scallop shells was characterized by a greater number of warm-water species.     

Epibiotic Bacteria on Several Ciliates from Marine Sediments

ABSTRACT Bacterial epibionts were observed on the surface of the marine sediment ciliate Geleia fossata. Rod-shaped bacteria, from 2-10 X10³ per ciliate, were universally positioned in ciliated grooves, in apparent spatial association with dikinetids. SEM and TEM examination of the ciliates confirmed that a tight affiliation exists between the epibiotic bacteria and ciliate cortex infrastructures. These observations, as well as the distinct bacterial distribution pattern over ciliate surface, suggest that there is a close epibiont/host physiological integration. Epibiotic bacteria were also observed on the surfaces of other sediment ciliates from the genera Loxophyllum, Tracheloraphis, Geleia, Paraspathidium , and Cyclidium. These findings indicate that the bacterial/protozoa associations are widespread in the marine benthic environment. The potential benefits for both epibionts and their hosts are discussed.