Population distribution,host-switching,and chemical sensing in the symbiotic shrimp <Emphasis Type="Italic">Lysmata pederseni</Emphasis>: implications for its mating system in a changing reef seascape

Lysmata pederseni, a protandric simultaneously hermaphroditic shrimp that inhabits the tube sponge Callyspongia vaginalis, is monogamous in the central and southeastern Caribbean Sea. We tested the null hypothesis of monogamy in a northern Caribbean population. In the Florida Keys, shrimps did not inhabit host individuals in pairs with a frequency greater than expected by chance alone. Hermaphrodites inhabited sponges solitarily and often brooded embryos. Hermaphrodites do not store sperm and need to be inseminated shortly after molting to fertilize a new batch of eggs. Thus, males and/or other hermaphrodites are likely switching among host individuals in search of sexual partners. Field experiments demonstrated low shrimp host fidelity. Host residence time was ~2 times shorter for males than for hermaphrodites. We inferred a polygynandrous mating system in L. pederseni from the Florida Keys, with male-role and young hermaphrodites often moving among sponges in search of older, more sedentary, female-role hermaphrodites. We expected shrimps to use water-borne chemical cues originating from conspecifics or sponges to locate sexual partners. Experiments demonstrated that shrimps were attracted to water-borne cues originating from sponges but not conspecifics. We have described the mating system of a reef-associated shrimp in a fast-pace shifting seascape increasingly dominated by sponges and vanishing stony corals. In the central and southeastern Caribbean Sea, with greater coral cover and lower sponge abundance than in the Florida Keys, the same species is monogamous. Whether or not similar shifts in the social organization of other coral reef-dwelling marine organisms are occurring due to contemporary changes in seascapes is a relevant topic that deserves further attention.     

Pyrosequencing Characterization of the Microbiota from Atlantic Intertidal Marine Sponges Reveals High Microbial Diversity and the Lack of Co-Occurrence Patterns

Sponges are ancient metazoans that host diverse and complex microbial communities. Sponge-associated microbial diversity has been studied from wide oceans across the globe, particularly in subtidal regions, but the microbial communities from intertidal sponges have remained mostly unexplored. Here we used pyrosequencing to characterize the microbial communities in 12 different co-occurring intertidal marine sponge species sampled from the Atlantic coast, revealing a total of 686 operational taxonomic units (OTUs) at 97% sequence similarity. Taxonomic assignment of 16S ribosomal RNA tag sequences estimated altogether 26 microbial groups, represented by bacterial (75.5%) and archaeal (22%) domains. Proteobacteria (43.4%) and Crenarchaeota (20.6%) were the most dominant microbial groups detected in all the 12 marine sponge species and ambient seawater. The Crenarchaeota microbes detected in three Atlantic Ocean sponges had a close similarity with Crenarchaeota from geographically separated subtidal Red Sea sponges. Our study showed that most of the microbial communities observed in sponges (73%) were also found in the surrounding ambient seawater suggesting possible environmental acquisition and/or horizontal transfer of microbes. Beyond the microbial diversity and community structure assessments (NMDS, ADONIS, ANOSIM), we explored the interactions between the microbial communities coexisting in sponges using the checkerboard score (C-score). Analyses of the microbial association pattern (co-occurrence) among intertidal sympatric sponges revealed the random association of microbes, favoring the hypothesis that the sponge-inhabiting microbes are recruited from the habitat mostly by chance or influenced by environmental factors to benefit the hosts.     

Biodiversity, host specificity, and dominance by eusocial species among sponge-dwelling alpheid shrimp on the Belize Barrier Reef

Alpheid shrimp represent an abundant and diverse, but poorly characterized, component of the cryptic biodiversity of coral reefs worldwide. Sponge‐inhabiting alpheids provide a promising model system for exploring patterns of cryptic reef biodiversity because their habitats (hosts) are discrete and qualitatively distinct units. We tabulated data from 14 years of collections at Carrie Bow Cay, Belize to quantify patterns of diversity, host specificity, and dominance among sponge‐dwelling shrimp (Synalpheus), with special attention to eusocial species. From > 600 sampled sponges of 17 species, we recognized at least 36 Synalpheus shrimp species. Of these, 15 (42%) were new to science. Species accumulation curves suggest that we have sampled most of the Synalpheus diversity at Carrie Bow Cay. Diversity of sponge‐dwelling Synalpheus was slightly higher in shallow water, probably because of greater habitat diversity, than in deep water. Host specificity was surprisingly high, with > 50% of all shrimp species found in only a single sponge species each, although some shrimp species used as many as six hosts. Cohabitation of individual sponges by multiple shrimp species was rarer than expected by chance, supporting previous distributional and behavioural evidence that competition for hosts is strong and moulds patterns of host association. The fauna of most well‐sampled sponge species was dominated, both in numbers of individuals and in frequency of occurrence, by eusocial species. Eusocial shrimp species also inhabited a significantly greater number of sponge species than did non‐social shrimp. Consequently, > 65% of shrimp in our quantitative samples belonged to the four eusocial species, and on a per‐species basis, eusocial species were 17 times as abundant as non‐social species. Our data suggest that the highly diverse sponge‐dwelling shrimp assemblage of the Belize Barrier Reef is structured by competition, and that eusociality has allowed a small number of species to dominate the sponge resource.     

Species boundaries, specialization, and the radiation of sponge-dwelling alpheid shrimp

Microevolutionary studies and natural history suggest that host-specialization has promoted the high diversity of tropical sponge-dwelling snapping shrimps (Decapoda, Alpheidae, Synalpheus ). Yet the taxonomic difficulty of this genus has precluded rigorous tests of this hypothesis. S. rathbunae Coutiére is among the most abundant invertebrates inhabiting the framework of sponges and dead coral that forms the floor of Caribbean coral reefs. Even within a small area S. rathbunae exhibits the apparently wide variation in size, color, and morphology that has long frustrated efforts to identify and define species boundaries within this large (> 100 described species) genus. Here I show that sympatric populations of this nominal species occupying different sponge hosts display clear, concordant differences in allozyme genotypes and in multivariate morphometries, confirming that the populations represent three distinct biological species. Moreover, careful field sampling revealed that the three S. rathbunce taxa and the closely related S. filidigitus Armstrong showed almost no overlap in the species of hosts occupied. Interestingly, while there was significant differentiation between Belizean and Panamanian populations of the one taxon that occurred at both sites (∼1500 km apart), these populations were recognizable as conspecific using both genetic and morphological characters. These results show that (1) diversity of Synalpheus , which is already among the most species-rich crustacean genera, is probably several times higher than currendy recognized, and (2) species of sponge-dwelling Synalpheus are highly host-specific, with related species distinctly segregated among hosts. Together with previous evidence of host race differentiation within shrimp species, these results suggest a primary role for resource specialization in the origin and/or maintenance of this group's characteristically high diversity.     

Influence of sponge morphology on the composition of the polychaete associated fauna from Rocas Atoll,northeast Brazil

The relationships between the morphology of sponges and variables describing their harbored polychaete fauna were analyzed along Rocas, the only known South Atlantic Atoll, together with the location on host and feeding habits. The identification and quantification of all the associated organisms highlighted the dominance of polychaetes. The adults of the symbiotic species Haplosyllis spongicola were the most dominant sponge endobionts. However, both juveniles and epitokes (reproductive individuals) of H. spongicola were also found, suggesting that this species completes its life cycle inside the host. Polychaete density was significantly greater in lobate sponges than in massive and encrusting forms. Conversely, the highly specific symbiotic mode of life of H. spongicola species seems to play a major role in structuring the composition of the polychaete fauna in relation to sponge morphotypes along Rocas Atoll.     

Biochemical Aspects of Parasitism by the Angiosperm Parasites: Phenolics in Parasites and Hosts

the contents of total phenolics in three parasitic angiosperms, Cuscuta species, Orobanche aegyptiaca and Dendrophthoe falcata and their respective hosts, were colorimetrieally determined. A biochemical comparison was made of the phenolics on the basis of the ability of alcoholic extracts of the tissues to inhibit amylose phosphorylase in vitro. High concentration of phenolics seemed to be a general feature of parasitic angiosperms. An increase in the concentration of the phenolics occurred in the tissues of infected hosts, in comparison with controls. the phenolics of Orobanche and mistletoe had inhibitory activity against amylose phosphorylase, but those of Cuscuta developed the inhibitory ability only when growing on hosts which themselves possessed inhibitory phenolics. the inhibitory activity of host phenolics was sometimes altered as a result of infection by parasite. Although the hosts often exerted some influence on the concentration and the inhibitory activity of phenolics in the parasites, there was no direct relationship between host and parasite phenolics. the sum of the phenolics in the tissues of parasite and the infected bost generally exceeded the phenolics in the tissues of the control host. The content of phenolics and their inhibitory activity did not appear to be directly related to the resistance of a host or to the extent of its susceptibility to parasite infection.     

Effects of three Caribbean cleaner shrimps on ectoparasitic monogeneans in a semi-natural environment

Most research on cleaning symbioses on coral reefs has focused on fish clients being cleaned by smaller fishes. While many shrimps and other crustaceans are reported as cleaners, whether they remove parasites from fish hosts and can effectively regulate populations of ectoparasites is unclear. The effects of Pederson shrimp (Periclimenes pedersoni), spotted shrimp (P. yucatanicus), and banded coral shrimp (Stenopus hispidus), on the parasitic monogenean Neobenedenia melleni on a host reef fish, blue tang (Acanthurus coeruleus), were investigated. The abundance and size of N. melleni from fish with and without access to shrimps in a semi-natural macrocosm was quantified. P. pedersoni had a strong effect on both the abundance and size of parasites. In contrast, P. yucatanicus and S. hispidus had no effect on the abundance of parasites but had a small yet statistically significant effect on average size. These data suggest that P. pedersoni can play a significant role in the biological regulation of at least some ectoparasites on Caribbean reef fishes, but further suggest that some other shrimps regarded as “cleaners” may have little or no effectiveness at removing parasites and underscore the need for further verification before this term is applied.     

Sandpipers Select Red Brine Shrimps Rich in Both Carotenoids and Parasites

Cestode parasites using brine shrimps Artemia as intermediate hosts and waterbirds as final hosts are known to change the colour of shrimps from pale pink to bright red owing to an increase in carotenoid pigments. In order to assess whether this coloration increases the chances of shrimp predation by the final host, we conducted an experiment in which 10 Calidris sandpipers were given a choice of 20 bright red and 20 pale pink shrimps. A strong preference for red, infected shrimps was recorded, suggesting that red colour is a manipulation by cestodes to increase transmission to its final hosts. The factors regulating access and functions of carotenoids in birds are under intense debate. For the first time, we show that access to carotenoid-rich foods in birds can have a cost derived from an increased exposure to parasites. To our knowledge, this is the first time that avian hosts have been shown to exert a colour-based preference for parasitized prey.     

Sponge-associated bacteria are strictly maintained in two closely related but geographically distant sponge hosts

The giant barrel sponges Xestospongia muta and Xestospongia testudinaria are ubiquitous in tropical reefs of the Atlantic and Pacific Oceans, respectively. They are key species in their respective environments and are hosts to diverse assemblages of bacteria. These two closely related sponges from different oceans provide a unique opportunity to examine the evolution of sponge-associated bacterial communities. Mitochondrial cytochrome oxidase subunit I gene sequences from X. muta and X. testudinaria showed little divergence between the two species. A detailed analysis of the bacterial communities associated with these sponges, comprising over 900 full-length 16S rRNA gene sequences, revealed remarkable similarity in the bacterial communities of the two species. Both sponge-associated communities include sequences found only in the two Xestospongia species, as well as sequences found also in other sponge species and are dominated by three bacterial groups, Chloroflexi, Acidobacteria, and Actinobacteria. While these groups consistently dominate the bacterial communities revealed by 16S rRNA gene-based analysis of sponge-associated bacteria, the depth of sequencing undertaken in this study revealed clades of bacteria specifically associated with each of the two Xestospongia species, and also with the genus Xestospongia, that have not been found associated with other sponge species or other ecosystems. This study, comparing the bacterial communities associated with closely related but geographically distant sponge hosts, gives new insight into the intimate relationships between marine sponges and some of their bacterial symbionts.     

Molecular detection of fungal communities in the Hawaiian marine sponges Suberites zeteki and Mycale armata

Symbiotic microbes play a variety of fundamental roles in the health and habitat ranges of their hosts. While prokaryotes in marine sponges have been broadly characterized, the diversity of sponge-inhabiting fungi has barely been explored using molecular approaches. Fungi are an important component of many marine and terrestrial ecosystems, and they may be an ecologically significant group in sponge-microbe interactions. This study tested the feasibility of using existing fungal primers for molecular analysis of sponge-associated fungal communities. None of the eight selected primer pairs yielded satisfactory results in fungal rRNA gene or internal transcribed spacer (ITS) clone library constructions. However, 3 of 10 denaturing gradient gel electrophoresis (DGGE) primer sets, which were designed to preferentially amplify fungal rRNA gene or ITS regions from terrestrial environmental samples, were successfully amplified from fungal targets in marine sponges. DGGE analysis indicated that fungal communities differ among different sponge species (Suberites zeteki and Mycale armata) and also vary between sponges and seawater. Sequence analysis of DGGE bands identified 23 and 21 fungal species from each of the two sponge species S. zeteki and M. armata, respectively. These species were representatives of 11 taxonomic orders and belonged to the phyla of Ascomycota (seven orders) and Basidiomycota (four orders). Five of these taxonomic orders (Malasseziales, Corticiales, Polyporales, Agaricales, and Dothideomycetes et Chaetothyriomcetes incertae sedis) have now been identified for the first time in marine sponges. Seven and six fungal species from S. zeteki and M. armata, respectively, are potentially new species because of their low sequence identity (< or =98%) with their references in GenBank. Phylogenetic analysis indicated sponge-derived sequences were clustered into "marine fungus clades" with those from other marine habitats. This is the first report of molecular analysis of fungal communities in marine sponges, adding depth and dimension to our understanding of sponge-associated microbial communities.     

繁茂膜海绵细胞内微生物多样性的研究

采用海绵组织离散、细胞分离的方法,对繁茂膜海绵细胞进行纯化、胞内微生物DNA提取,构建了繁茂膜海绵细胞内微生物的16SrDNA克隆,对其遗传多样性进行了分析,发现海绵细胞内微生物16SrDNA序列主要归类于紫硫细菌门(Proteobacteria)中的α-亚门、γ-亚门和浮霉菌门(Planctomycetes)等类群。与研磨直接提取海绵组织DNA所得海绵组织中总微生物多样性相比,海绵细胞内存在丰富的浮霉菌(23%),说明浮霉菌主要存在于海绵细胞胞内。     

Ultrastructure, Molecular Phylogenetics, and Chlorophyll a Content of Novel Cyanobacterial Symbionts in Temperate Sponges

Marine sponges often harbor photosynthetic symbionts that may enhance host metabolism and ecological success, yet little is known about the factors that structure the diversity, specificity, and nature of these relationships. Here, we characterized the cyanobacterial symbionts in two congeneric and sympatric host sponges that exhibit distinct habitat preferences correlated with irradiance: Ircinia fasciculata (higher irradiance) and Ircinia variabilis (lower irradiance). Symbiont composition was similar among hosts and dominated by the sponge-specific cyanobacterium Synechococcus spongiarum. Phylogenetic analyses of 16S-23S rRNA internal transcribed spacer (ITS) gene sequences revealed that Mediterranean Ircinia spp. host a specific, novel symbiont clade ("M") within the S. spongiarum species complex. A second, rare cyanobacterium related to the ascidian symbiont Synechocystis trididemni was observed in low abundance in I. fasciculata and likewise corresponded to a new symbiont clade. Symbiont communities in I. fasciculata exhibited nearly twice the chlorophyll a concentrations of I. variabilis. Further, S. spongiarum clade M symbionts in I. fasciculata exhibited dense intracellular aggregations of glycogen granules, a storage product of photosynthetic carbon assimilation rarely observed in I. variabilis symbionts. In both host sponges, S. spongiarum cells were observed interacting with host archeocytes, although the lower photosynthetic activity of Cyanobacteria in I. variabilis suggests less symbiont-derived nutritional benefit. The observed differences in clade M symbionts among sponge hosts suggest that ambient irradiance conditions dictate symbiont photosynthetic activity and consequently may mediate the nature of host-symbiont relationships. In addition, the plasticity exhibited by clade M symbionts may be an adaptive attribute that allows for flexibility in host-symbiont interactions across the seasonal fluctuations in light and temperature characteristic of temperate environments.     

Avian cestodes affect the behaviour of their intermediate host Artemia parthenogenetica: an experimental study

The brine shrimp Artemia parthenogenetica (Crustacea, Branchiopoda) is intermediate host for several cestode species whose final hosts are waterbirds. Previous field studies have shown that brine shrimps infected with cestodes have a bright red colour and are spatially segregated in the water column. However, the ethological mechanisms explaining such field observations are unknown. Changes in appearance and behaviour induced by trophically transmitted parasites have been shown to increase the risk of predation by the final host. In this experimental study, we compared the behaviour of uninfected Artemia and those infected by avian cestodes. We found that parasitised individuals behave differently from unparasitised ones in several ways. In contrast to uninfected individuals, infected brine shrimps were photophilous and showed increased surface-swimming behaviour. These observations suggest that the modified behaviour (in addition to the bright red colour of the majority of the infected individuals) results in infected brine shrimps becoming more vulnerable to avian final hosts, which facilitates parasite transmission. We discuss our results in terms of the adaptive nature of behavioural changes and their potential implications for the hypersaline ecosystem.     

Chymase as a proangiogenic factor. A possible involvement of chymase-angiotensin-dependent pathway in the hamster sponge angiogenesis model

We investigated the profound involvement of chymase, an alternative angiotensin II-generating enzyme, in angiogenesis using a hamster sponge implant model. In vivo transfection of human pro-chymase cDNA or a direct injection of purified chymase into the sponges implanted resulted in marked increment of hemoglobin contents in the sponge granuloma tissues, demonstrating that chymase has an ability to elicit angiogenesis and is a potent angiogenic factor. Daily injection of basic fibroblast growth factor into the sponges implanted also induced angiogenesis, which was suppressed by the treatment with chymostatin, an inhibitor of chymase, or TCV-116, an antagonist of angiotensin II (Ang II) type 1 receptor. Expression of chymase mRNA and production of Ang II in the granuloma tissues were enhanced by the stimulation with basic fibroblast growth factor. Chymase activity in the sponge granulomas increased in parallel with the rise in hemoglobin contents, and mast cells observed in the granuloma tissues were positively stained with anti-chymase antibody. Exogenous administration not only of Ang II but of angiotensin I (Ang I) directly into the sponges could enhance angiogenesis. Chymostatin inhibited the angiogenesis induced by Ang I but not Ang II, suggesting the presence of a chymase-like Ang II-generating activity in the sponge granulomas. Our results may suggest a potential ability of chymase to promote angiogenesis through the local chymase-dependent and angiotensin-converting enzyme-dependent Ang II generating system in pathophysiological angiogenesis.     

Ecological generalism facilitates the evolution of sociality in snapping shrimps

Evidence from insects and vertebrates suggests that cooperation may have enabled species to expand their niches, becoming ecological generalists and dominating the ecosystems in which they occur. Consistent with this idea, eusocial species of sponge‐dwelling Synalpheus shrimps from Belize are ecological generalists with a broader host breadth and higher abundance than non‐eusocial species. We evaluate whether sociality promotes ecological generalism (social conquest hypothesis) or whether ecological generalism facilitates the transition to sociality (social transition hypothesis) in 38 Synalpheus shrimp species. We find that sociality evolves primarily from host generalists, and almost exclusively so for transitions to eusociality. Additionally, sponge volume is more important for explaining social transitions towards communal breeding than to eusociality, suggesting that different ecological factors may influence the independent evolutionary origins of sociality in Synalpheus shrimps. Ultimately, our results are consistent with the social transition hypothesis and the idea that ecological generalism facilitates the transition to sociality.     

Lifestyle of sponge symbiont phages by host prediction and correlative microscopy

Bacteriophages (phages) are ubiquitous elements in nature, but their ecology and role in animals remains little understood. Sponges represent the oldest known extant animal-microbe symbiosis and are associated with dense and diverse microbial consortia. Here we investigate the tripartite interaction between phages, bacterial symbionts, and the sponge host. We combined imaging and bioinformatics to tackle important questions on who the phage hosts are and what the replication mode and spatial distribution within the animal is. This approach led to the discovery of distinct phage-microbe infection networks in sponge versus seawater microbiomes. A new correlative in situ imaging approach (‘PhageFISH-CLEM‘) localised phages within bacterial symbiont cells, but also within phagocytotically active sponge cells. We postulate that the phagocytosis of free virions by sponge cells modulates phage-bacteria ratios and ultimately controls infection dynamics. Prediction of phage replication strategies indicated a distinct pattern, where lysogeny dominates the sponge microbiome, likely fostered by sponge host-mediated virion clearance, while lysis dominates in seawater. Collectively, this work provides new insights into phage ecology within sponges, highlighting the importance of tripartite animal-phage-bacterium interplay in holobiont functioning. We anticipate that our imaging approach will be instrumental to further understanding of viral distribution and cellular association in animal hosts.Subject terms: Bacteriophages, Microbial ecology, Symbiosis     

Cryptic diversity of the symbiotic cyanobacterium Synechococcus spongiarum among sponge hosts

Cyanobacteria are common members of sponge-associated bacterial communities and are particularly abundant symbionts of coral reef sponges. The unicellular cyanobacterium Synechococcus spongiarum is the most prevalent photosynthetic symbiont in marine sponges and inhabits taxonomically diverse hosts from tropical and temperate reefs worldwide. Despite the global distribution of S. spongiarum , molecular analyses report low levels of genetic divergence among 16S ribosomal RNA (rRNA) gene sequences from diverse sponge hosts, resulting either from the widespread dispersal ability of these symbionts or the low phylogenetic resolution of a conserved molecular marker. Partial 16S rRNA and entire 16S–23S rRNA internal transcribed spacer (ITS) genes were sequenced from cyanobacteria inhabiting 32 sponges (representing 18 species, six families and four orders) from six geographical regions. ITS phylogenies revealed 12 distinct clades of S. spongiarum that displayed 9% mean sequence divergence among clades and less than 1% sequence divergence within clades. Symbiont clades ranged in specificity from generalists to specialists, with most (10 of 12) clades detected in one or several closely related hosts. Although multiple symbiont clades inhabited some host sponges, symbiont communities appear to be structured by both geography and host phylogeny. In contrast, 16S rRNA sequences were highly conserved, exhibiting less than 1% sequence divergence among symbiont clades. ITS gene sequences displayed much higher variability than 16S rRNA sequences, highlighting the utility of ITS sequences in determining the genetic diversity and host specificity of S. spongiarum populations among reef sponges. The genetic diversity of S. spongiarum revealed by ITS sequences may be correlated with different physiological capabilities and environmental preferences that may generate variable host–symbiont interactions.     

Deep sequencing reveals exceptional diversity and modes of transmission for bacterial sponge symbionts

Marine sponges contain complex bacterial communities of considerable ecological and biotechnological importance, with many of these organisms postulated to be specific to sponge hosts. Testing this hypothesis in light of the recent discovery of the rare microbial biosphere, we investigated three Australian sponges by massively parallel 16S rRNA gene tag pyrosequencing. Here we show bacterial diversity that is unparalleled in an invertebrate host, with more than 250,000 sponge-derived sequence tags being assigned to 23 bacterial phyla and revealing up to 2996 operational taxonomic units (95% sequence similarity) per sponge species. Of the 33 previously described 'sponge-specific' clusters that were detected in this study, 48% were found exclusively in adults and larvae - implying vertical transmission of these groups. The remaining taxa, including 'Poribacteria', were also found at very low abundance among the 135,000 tags retrieved from surrounding seawater. Thus, members of the rare seawater biosphere may serve as seed organisms for widely occurring symbiont populations in sponges and their host association might have evolved much more recently than previously thought.