Crinoid phylogeny: a preliminary analysis (Echinodermata: Crinoidea)

We describe the first molecular and morphological analysis of extant crinoid high-level inter-relationships. Nuclear and mitochondrial gene sequences and a cladistically coded matrix of 30 morphological characters are presented, and analysed by phylogenetic methods. The molecular data were compiled from concatenated nuclear-encoded 18S rDNA, internal transcribed spacer 1, 5.8S rDNA, and internal transcribed spacer 2, together with part of mitochondrial 16S rDNA, and comprised 3,593 sites, of which 313 were parsimony-informative. The molecular and morphological analyses include data from the bourgueticrinid Bathycrinus; the antedonid comatulids Dorometra and Florometra; the cyrtocrinids Cyathidium, Gymnocrinus, and Holopus; the isocrinids Endoxocrinus, and two species of Metacrinus; as well as from Guillecrinus and Caledonicrinus, whose ordinal relationships are uncertain, together with morphological data from Proisocrinus. Because the molecular data include indel-rich regions, special attention was given to alignment procedure, and it was found that relatively low, gene-specific, gap penalties gave alignments from which congruent phylogenetic information was obtained from both well-aligned, indel-poor and potentially misaligned, indel-rich regions. The different sequence data partitions also gave essentially congruent results. The overall direction of evolution in the gene trees remains uncertain: an asteroid outgroup places the root on the branch adjacent to the slowly evolving isocrinids (consistent with palaeontological order of first appearances), but maximum likelihood analysis with a molecular clock places it elsewhere. Despite lineage-specific rate differences, the clock model was not excluded by a likelihood ratio test. Morphological analyses were unrooted. All analyses identified three clades, two of them generally well-supported. One well-supported clade (BCG) unites Bathycrinus and Guillecrinus with the representative (chimaeric) comatulid in a derived position, suggesting that comatulids originated from a sessile, stalked ancestor. In this connection it is noted that because the comatulid centrodorsal ossicle originates ontogenetically from the column, it is not strictly correct to describe comatulids as unstalked crinoids. A second, uniformly well-supported clade contains members of the Isocrinida, while the third clade contains Gymnocrinus, a well-established member of the Cyrtocrinida, together with the problematic taxon Caledonicrinus, currently classified as a bourgueticrinid. Another cyrtocrinid, Holopus, joins this clade with only weak molecular, but strong morphological support. In one morphological analysis Proisocrinus is weakly attached to the isocrinid clade. Only an unusual, divergent 18S rDNA sequence was obtained from the morphologically strange cyrtocrinid Cyathidium. Although not analysed in detail, features of this sequence suggested that it may be a PCR artefact, so that the apparently basal position of this taxon requires confirmation. If not an artefact, Cyathidium either diverged from the crinoid stem much earlier than has been recognised hitherto (i.e., it may be a Palaeozoic relic), or it has an atypically high rate of molecular evolution.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by J.P. Thorpe, Port Erin     

Observations on the food of Antedon bifida (Echinodermata: Crinoidea)

The gut contents of specimens of the temperate crinoid Antedon bifida (Pennant) were collected from Dalkey, County Dublin, at monthly intervals for 1 yr, and occasionally from other areas around the Irish coast. Quantitative analysis showed the main components to be detritus (65%), plankton (18%) and inorganic particles (17%). This composition was remarkably constant throughout the year and at all locations, the main variation being in the plankton which was at a minimum in February (4%) and reached a maximum in August (36%). Food particle collection appeared to be nonselective within the appropriate size group, the essential feature of a particle being that it would fit into the ambulacral groove. Fragile planktonic diatoms and dinoflagellates, which occurred in blooms in summer and autumn, were digested by A. bifida, but more robust diatoms passed intact through the gut. Histochemical investigations indicate that the bulk of the detritus is refractory, and it is suggested that its associated micro-flora and micro-fauna may be a major source of nutriment for this crinoid.     

Food composition of crinoids (Crinoidea: Echinodermata) in relation to stalk length and fan density: their paleoecological implications

Crinoids have been diverse organisms in marine epifaunal filter feeding communities at any level of tiering above the substrate since they appeared in the Ordovician. Feeding is regarded as the most important factor in producing the crinoid tiering, which is primarily defined by stalk length. The gut contents of five sympatric crinoid species (three isocrines and two comatulids) were observed, and these were compared with the stalk length and the fan density. We have classified these crinoid species into four groups based on the stalk length and fan density, e.g., long stalk with low fan density, long stalk with high fan density, short stalk with low fan density, and short stalk with high fan density. In the gut contents, diatom crusts were found mainly from species with longer stalks, and chlorophyll-like fluorescent material were only detected from the groups with a shorter or no stalk. The group with lower fan density contained more inorganic particles than the group with higher fan density. Therefore, the gut contents and their amounts depend on their stalk lengths and their fan densities. The results imply that diversified morphologies in the crinoids have evolved through adaptations to different ecological factors such as difference in their diets.     

Reproductive biology of the common antarctic crinoid Promachocrinus kerguelensis (Echinodermata: Crinoidea)

Histological examinations of the gonads of Promachocrinus kerguelensis Carpenter, 1888 collected in November 1984 from McMurdo Sound, Antarctica, revealed ripe ovaries containing three distinct size classes of oocytes: 20 to 90, 91 to 150 and 151 to 180 m diameter as well as ova. Testes were ripe, and filled with mature sperms. There was marked inter-pinnule and inter-individual synchrony of gametogenesis and spawning is probably synchronous, at least within the sub-population sampled. The number of eggs per pinnule is allometrically related to body size. Body weight and arm length are positively correlated with total egg number. Annual fecundity was estimated to be 29 000 eggs (full-grown oocytes and ova) for a crinoid weighing 53.6 g wet weight with an arm length of 16.3 cm. The eggs float. Female genital pinnules contain more lipid and energy than male pinnules. Reproductive output and effort (23.4 to 65.2 kJ and 26 to 38%, respectively) increase with body size and are similar to values reported for other echinoderms with lecithotrophic larvae. The greatest overall energetic contribution is represented by the arms and genital pinnules. Spawning in P. kerguelensis probably occurs in November and December within McMurdo Sound. If development is slow, as in all other antarctic echinoderms studied to date, then settlement most likely occurs 2 to 3 mo later, after the short summer period of high phytoplankton productivity. High fecundity, a pelagic mode of reproduction, and the high probability of a slow rate of development may contribute to the wide geographic distribution of this common antarctic crinoid.     

Digestive mechanics and gluttonous feeding in the feather starOligometra serripinna (Echinodermata: Crinoidea)

The movement and digestion of food in the gut ofOligometra serripinna (Carpenter) were studied at Lizard Island (14°3842S; 145°2710E) in the austral winter of 1986. Feather stars in the laboratory were fed a brief, small meal of brine shrimp nauplii and killed at increasing time intervals thereafter. Histological reconstructions showed that the ingested nauplii progressed along the digestive tract surprisingly quickly. Some nauplii were found in the mid and hind intestine in only 30 min, and all of the nauplii had reached the hind intestine and rectum in 1 h. Digestion of the nauplii had started at 1 h, and only a few fragments of naupliar exoskeleton remained in the hind intestine and rectum 5 h after the start of feeding. Videotape analysis showed that no fecal pellets were released during this experiment. In the natural environment ofO. serripinna, ingested particles may similarly be transported quickly to the hind part of the gut and digested there — when feather stars were fixed in the field, most of the gut contents were found in the hind intestine and rectum.O. serripinna, which efficiently rejects inert particles before they are ingested, usually defecates infrequently (probably not more than once over a span of many hours) and differs from some other feather stars that ingest numerous inert particles and defecate much more frequently. When specimens ofO. serripinna were fed continuously on brine shrimp nauplii,Artemia sp. (San Francisco strain), in the laboratory, the feather stars fed gluttonously, packing their guts with several hundred nauplii in 1 to 2 h. Thereafter, superfluous feeding began (i.e., further ingestions appeared to force undigested nauplii, some of them still living, out of the anus). These observations suggest thatO. serripinna usually feeds at relatively modest rates in its natural habitat, but can feed gluttonously to take advantage of infrequent patches of highly concentrated, nutritious particles (e.g. copepod swarms, migrating demersal zooplankton, and invertebrate gametes from mass spawnings). It is likely that such patches of nutritious particles are usually small enough to drift out of reach of the feather stars before gluttonous feeding proceeds to superfluous feeding. Opportunities for superfluous feeding in nature are probably very infrequent (e.g. ingestion of coral gametes and embryos after a mass spawning), and the feather stars evidently have no behavior that stops further ingestions after the gut becomes filled to capacity.     

Reproductive biology and early life history of the hermaphroditic feather star Dorometra sesokonis (Echinodermata: Crinoidea)

Reproduction and larval/post-larval development of the one of the smallest known comatulid Dorometra sesokonis were studied on the coral reefs of Sesoko-jima Island, Okinawa, Japan. Breeding individuals were found every month from May 2003 to April 2004 indicating continuous reproduction. Individuals possessed both mature ovaries and testes, and were therefore characterized as simultaneous hermaphrodites. This species was observed to be an external brooder in which the embryos developed on the surface of parental genital pinnules until the doliolaria larval stage, and subsequently released doliolariae settled on substrata within 4 days after hatching. Larvae then metamorphosed into the post-larval cystidean stage, which lasted for more than a month. The reproductive features of this species are unique among crinoids, and, together with juvenile morphological features observed in adults, can be interpreted as adaptations to their cryptic habitat, which is subject to frequent natural disturbances.     

Effects of current speed on filtration during suspension feeding in Oligometra serripinna (Echinodermata: Crinoidea)

Suspension feeding by the crinod Oligometra serripinna was studied at Lizard Island, Australia, in 1986. Video recordings were made of 90-m particles interacting with the filter of the crinoid in a laboratory flow chamber. A complete census of particles was possible because both the capture event and the filter area could be defined unequivocally. Also, because O. serripinna is a passive suspension feeder, a census of partcles could be made at different ambient current speeds without interference due to active pumping by the crinoid. Experiments were run at seven current speeds from 0.9 to 13.3 cm s^-1. Particles approaching the filter: (1) were captured, (2) passed through the filter without triggering a capture event, (3) passed through the filter after escaping from an unsuccessful capture event, or (4) were deflected around the filter. With increasing current speed, the proportion of deflections declined and the proportion of particles passing through rose: these results could be partially explained by the progressive widening of the spaces within the filter due to distortion of filter parts by the current. The proportion of captures (normalized to approaches) was comparatively low at 0.9 cm s^-1, rose to a relatively constant maximum from 1.7 to 6.4 cm s^-1, and then declined progressively at 9.5 and 13.3 cm s^-1. These proportions were translated into capture rates for whole crinoids by taking into consideration both the encounters with particles and the reduction of filter area by distortion of body parts at higher speeds. When plotted against current speed, capture rate peaked at 6.4 cm s^-1, which was close to the mean current speed that we measured on the reef in the microhabitat of O. serripinna.     

Reproductive cycle of the sea urchin Holopneustes purpurascens (Temnopleuridae: Echinodermata)

This study documents the annual reproductive cycle of the Temnopleurid echinoid Holopneustes purpurascens near Sydney. H. purpurascens reached sexual maturity between 11 and 26 mm diameter and was gonochoristic. The sequence of oogenesis was similar to previously described echinoids. However, the proportion of eggs at each stage showed no temporal pattern in abundance, suggesting that vitellogenesis is continuous throughout the year. Spermatogenesis was also similar to other echinoids and showed a significant increase in the proportion of partially spawned testes in spring, followed by a similar increase in the proportion of partially spawned testes in late spring and summer. Gonad indices showed a peak in weight from late winter to mid-spring (9.10ǃ.47 for females; 6.28ǂ.43 for males; mean-SE), then a decline for both sexes. Although spawning was variable over time, there was a peak in the proportion of spawning from late winter to early summer that correlated with the observed peaks in gonad indices and gametogenic cycles. The demographic implications of this reproductive pattern are that new recruits can enter the population through synchronous spawning during the breeding period, and smaller haphazard spawning events at other times of the year.     

Spawning behaviors of northeast Pacific sea cucumbers (Holothuroidea: Echinodermata)

Spawning behaviors observed from May 1981 through August 1983 in the laboratory and field are described and compared for twelve species of northeast Pacific holothuroids collected from waters of the San Juan Archipelago, Washington. For many species, elevation of the anterior end, stretching of the introvert, tentacle waving, and swaying are behaviors commonly associated with gamete release. Less common activities include waving of introvert podia, waves of body wall contractions, errant males, aggregations, and pseudocopulation. Positively buoyant eggs of four species [Psolus chitonoides Clark, Cucumaria miniata (Brandt), C. piperata (Stimpson), and C. fallax (Ludwig)] were packaged in pellets, ropes, or strands and released passively; a female Molpadia intermedia (Ludwig) discharged negatively buoyant eggs in a single explosive burst. Two externally brooding sea cucumbers [C. lubrica (Clark) and C. pseudocurata Deichmann] produced strands of dilution-resistant sperm that were carried by currents to nearby females. Three species commonly sympatric [P. chitonoides, C. miniata, and Eupentacta quinquesemita (Selenka)] in high-current rocky habitats have overlapping spring breeding seasons, but spawn at different hours of the day.Please address all requests for reprints to Dr. McEuen at the Harbor Branch Oceanographic Institution     

On the distribution and variability of the Indo-Pacific sea star Asterina wega (Echinodermata: Asteroidea) in the Mediterranean sea

The distribution of the Mediterranean populations of Asterina wega Perrier is described. Four populations were found to differ in size of animals and regeneration rate. The possible migratory pattern is duscussed.     

Reproductive cycles of tropical sea cucumbers (Echinodermata: Holothuroidea) in southern Taiwan

Reproductive cycles of Afrocucumis africana (Dendrochirotida), Actinopyga echinites, Holothuria leucospilota, H. cinerascens, H. difficilis (Aspidochirotida), Synaptamaculata, Opheodesoma grisea, Patinapta taiwaniensis and Polycheira rufescens (Apodida), representing three orders of intertidal holothurians in southern Taiwan, were determined by gonad index and histological examination from March 1990 through September 1991. All nine species of holothurians have annual spawning periods lasting 2 to 4 mo in spring or summer. The early spawning of H. cinerascens and P. rufescens (suspension feeders) suggests that their feeding mode is related to food resources and may influence the reproductive period. In the direct-developing dendrochirotid (suspension feeder), gonad development occurs in cool months, and gametes are released at the beginning of warm months. In indirect-developing aspidochirotid and apodid (deposit feeder), gamete release occurs in late spring or summer and appears to be correlated with summer phytoplankton growth. The holothurians in the present study display spawning periodicity and duration similar to those of temperate species. Four species of holothurians lengthened spawning season with decreased latitude.     

L-Lactate dehydrogenase in the longitudinal muscle of the sea cucumber Sclerodactyla briareus (Echinodermata: Holothuroidea)

The infaunal holothurian Sclerodactyla briareus (Thyone briareus) is able to tolerate exposure to hypoxic conditions for over 2 days. Since the in vitro anaerobic degradation of glucose-U-C¹⁴ by longitudinal muscle preparations leads to an accumulation of labeled lactic acid, it is apparent that lactate dehydrogenase plays a key role during anoxia. Disc electrophoresis resolved one major band of lactate dehydrogenase (LDH) activity in longitudinal muscle extracts. Muscle extracts of s. briareus oxidized L-lactate only, as is the case with most other echinoderms. The apparent Michaelis constants for lactate oxidation and pyruvate reduction are dissimilar from values reported for other marine invertebrates. The LDH activity in this holothurian is higher than the activities reported for other marine invertebrates with the exception of some active, large arthropods. Substrate inhibition of LDH maximal activity occurs at substrate concentrations (pyruvate) exceeding 10 mM, and then to only a minor extent. The characteristics of lactate dehydrogenase from the longitudinal muscle of S. briareus support the hypothesis that it functions to maintain glycolytic flux during prolonged periods of exposure to anoxic or hypoxic conditions.     

Reproduction of the commercial sea cucumber Holothuria scabra (Echinodermata: Holothuroidea) in the Solomon Islands

Over a 3-year period (1996-1998), reproduction of the commercial sea cucumber Holothuria scabra (Jaeger, 1833) was investigated in the Solomon Islands to determine the spawning pattern and whether gametogenesis is continuous or seasonal. The gonad consisted of a single cohort of tubules that developed uniformly. Macroscopic examination of the gonads revealed that mature gametes were present throughout the year. Individuals with gonads at different stages of maturity were present in most samples. Partly spawned gonads were prevalent in females, whereas mature gonads were prevalent in males. The time at which the peak gonad index was recorded differed among years. Although gametogenesis was continuous, with a potential for prolonged gamete release, a period of enhanced spawning occurred during the dry season, from September to December. Maximum gonad indices were reached prior to and during this period of enhanced spawning. Histology revealed that gametogenesis reinitiated in partly spawned gonads, resulting in the presence of gametes at different stages of development in the gonad. The uniform growth of gonad tubules indicated that H. scabra does not conform to the progressive tubule recruitment model described for holothurian oogenesis. Continuous reproduction in H. scabra and prolonged availability of mature gametes would facilitate use of this species for aquaculture.     

Chemical defense in the eggs and embryos of antarctic sea stars (Echinodermata)

Fifteen species of reproductively mature echinoderms (11 sea stars, 3 sea urchins, 1 sea cucumber) were collected from McMurdo Sound, Antarctica, during austral spring and summer of 1985 and 1986; eggs and embryos were obtained, and were tested for ichthyonoxicity using the common marine killifishFundulus grandis as a model predator. Chemical deterrents occurred in the large, yolky eggs of the pelagic lecithotrophic sea starPerknaster fuscus and the planktotrophic sea starPorania antarctica. Brooded embryos of the sea starsDiplasterias brucei andNotasterias armata were also noxious. Significant ichthyonoxicity was not detected in the remaining 7 species of sea stars, 3 sea urchins, and 1 sea cucumber. Chemical deterrents were generally effective at concentrations below a single egg or embryo per agar test-pellet. Although chemicals found in these eggs and embryos are noxious to an allopatric fish, they may not be noxious to sympatric fish.     

Costs of extended brood protection in the Antarctic sea star, Neosmilaster georgianus (Echinodermata: Asteroidea)

Changes in size, morphology, and biochemical composition in adults and embryos of a brooding sea star, Neosmilaster georgianus (Sladen), were studied in a population adjacent to Anvers Island, Antarctic Peninsula (64°46′S; 64°04′W) during the austral spring, 1991. Five morphological stages of development were designated in 24 broods, and for each the weight and biochemical composition of the brooding adults and their embryos were determined. Between Stage 1 and 2, the dry weight (dw) and organic weight (ow) of the embryo did not change. From Stage 2 to 3, the dw and ow increased significantly by 10%. Stage 2 and 3 embryos were in clusters of a few (2 to 10) to as many as 40 individuals. In the smaller clusters, individual embryos were attached by tissue cords to another, sometimes atrophied, brood member. In the larger clusters, they were attached to a central mass of tissue containing remnants of embryos. We interpret these interactions as a form of cannibalism which may account for the weight gains between Stage 2 and 3. During Stages 4 and 5, as juvenile form was approached, the dw and ash weight of the young increased significantly and the ow decreased significantly. The calculated energy content for the juvenile (Stage 5) was not significantly different from the energy content of the earliest undifferentiated stage (Stage 1), an indication that most of the organic matter in the egg is the primary contribution to the large juvenile. In brooding females, pyloric caeca indices declined by 52% from Stage 1 to Stage 5 and pyloric energy stores declined by 63% due to proportionately equivalent declines in protein and lipid. The ovary index was low and increased only slightly during brood protection, while the size of the largest oocytes remained approximately 23% that of ova. Energy stores in the pyloric caeca of brooding N. georgianus thus become depleted over a long period of incubation during which the adults apparently starve. This may delay oocyte development and ultimately limit the frequency of individual reproduction. Received: 15 September 1998 / Accepted: 9 March 1999     

Karyotype analysis of the sea urchinParacentrotus lividus (Echinodermata): evidence for a heteromorphic chromosome sex mechanism

A consistent diploid number of 2n = 36 was determined for the sea urchinParacentrotus lividus from the Gulf of Palermo by analysis of mitotic chromosomes of both early developing embryos and male gonads. The haploid numbern = 18 was determined by counts of spermatocyte bivalents at diakinesis. A heteromorphic chromosome sex mechanism of the XY type is likely present in this species. This is indicated by the occurrence of a chromosomal pair, pair No. 2, which is heteromorphic in both morphology and size in about 50% of the mitotic figures (metaphases and anaphases) of einbryos. In addition, heteromorphism of the same pair of chromosomes occurred during spermatogonial metaphases in the five male specimens investigated here. The detection of a low chromosome number (2n = 36) compared to other echinoids (2n = 42 to 44), a heteromorphic chromosome sex system and the involvement of three chromosome pairs in nucleolar organization (NORs) provide evidence of the specialization of theP. lividus karyotype.     

Reproductive cycle of two commercial species of sea cucumber (Echinodermata: Holothuroidea) from Caribbean Panama

The reproductive status of the holothuroid species Isostichopus badionotus (Selenka, 1867) and Holothuria mexicana (Ludwig, 1875) was studied over 16 months in Bocas del Toro (Panama), from November 1999 to February 2001. Sexual reproduction was evaluated by the gonad index method, and by histology of gonad development. In addition, population structure was assessed based on sex ratio, minimum reproductive size, and length and weight distributions of males and females. The sex ratio in both species was 1:1, with a unimodal population distribution composed mainly of mature individuals. The minimum reproductive length and weight were 13-20 cm and 150 g, respectively, for both species, although reproductive individuals 10 cm in length were also found. A consistently higher gonad index was observed in H. mexicana, due to a high proportion of mature females and males and high gonad indices in most monthly samples. Gametogenesis and spawning patterns seemed to occur throughout the year, with periods of enhanced activity. Two periods of maximum reproductive activity were tentatively identified: July-November for I. badionotus and February-July for H. mexicana, but neither species had a single, sharply defined annual spawning event. Further work on these exploited holothuroids should examine the relationships between reproduction and environmental factors and between reproductive status and recruitment.     

Mass spawning by the sea urchin Evechinus chloroticus (Echinodermata: Echinoidea) in a New Zealand fiord

A mass synchronous spawning of the sea urchin Evechinus chloroticus (Valenciennes) was observed in situ in Doubtful Sound, a large New Zealand fiord. Spawning occurred between 17:30 hrs and 18:30 hrs on 27 January 1994 and coincided with a full moon, spring tides and a period of decreasing sea temperatures. During spawning, the sea urchins formed a dense spawning aggregation of both sexes, with >90% of the urchins observed spawning at the time. Spawned gametes clouded the water column, and some were eaten by small labrid fish species. The spawning, which may have been as widespread as 40 km, marked a 42 to 50% decrease in gonad indices and resulted in a widespread, dense cohort of  E. chloroticus larvae within the fiord. Received: 25 September 1997 / Accepted: 6 March 1998     

Acid-base status in the sea urchins Psammechinus miliaris and Echinus esculentus (Echinodermata: Echinoidea) during emersion

The acid-base status of two sea urchins, Psammechinus miliaris (Gmelin) and Echinus esculentus (L.) during experimental emersion has been investigated. Sea urchins were collected from the Firth of Clyde between August and September 1987. The carbon dioxide capacity of the coelomic fluid of P. miliaris was greater than that of E. esculentus, although both were low and only marginally greater than that of sea water. The pH of the coelonic fluid was also low (7.05 to 7.17) and was influenced mainly by the internal partial pressure of CO₂ (P_{CO 2}). Acid-base disturbance in the coelomic fluid of both species during emersion, although minimal, was more pronounced in E. esculentus than in P. miliaris and was due primarily to an increase in the internal P_{CO 2}, although there was an increase in the concentration of L-lactate in the coelomic fluid of E. esculentus. The coelomic fluid of both species was in a state of perfectly compensated respiratory acidosis. An increase in the concentration of divalent ions (Ca²⁺ and Mg²⁺) may be related to the dissolution of the test as a source of carbonate buffer.