Autoradiographic studies on sugar transport in the sea cucumber Holothuria scabra

Studies on the nutritional transport of holothurians have failed to delineate the role of the amoebocytes as well as the haemal system in this function. Autoradiographic studies have been carried out on tissues of animals injected with ¹⁴C glucose in order to localise the incorporation of the labelled sugar in tissues such as the alimentary canal, haemal system, respiratory tree and the body wall. In all tissues the amoebocytes were clearly labelled, indicating a key role in the transportation of sugars. Furthermore, it has been shown that sugar also enters the tissues by diffusion. It is interesting that the haemal system appears to transport glucose more rapidly than other tissues as indicated by the heavy labelling.     

Reproductive biology of the commercial sea cucumber Holothuria fuscogilva in the Solomon Islands

 Reproduction of Holothuria fuscogilva (Selenka, 1867) in the Solomon Islands was investigated over a 4 yr period (1994 to 1998) by macroscopic and microscopic examination of the gonad tubules, the gonad index (GI) method, histological examination of gametogenesis, and spawning-induction trials. The gonad consisted of numerous tubules that dominated the coelom of gravid specimens. New tubules appeared in March, and grew in size and extent of branching until they reached their maximum size and maturity in August. Spawning occurred from August to October, with the majority of gametes released during October, although it was only partial in many individuals. After spawning, the tubules appeared wrinkled and resorbed into the gonad basis. A five-stage gonad maturity scale based on the macroscopic appearance of the gonad tubules corresponded with discrete stages of gametogenesis identified by histology. Gametogenesis was initiated in mid-March, with oogenesis and spermatogenesis occurring in parallel, followed by the growing stage (May to July) which was marked by active gamete development. Successful induction of spawning during the breeding period corroborated the GI and histological data. The uniform growth of gonad tubules indicated that H. fuscogilva in the Solomon Islands does not conform to the progressive tubule recruitment model described for other holothurians. An important application of this study is that the appearance of gonad tubules, removed by biopsy, can be used to determine the gonad condition of wild adults or captive broodstock. Received: 28 June 1999 / Accepted: 8 March 2000     

Opsonin-like molecule found in coelomic fluid of a sea cucumber, Holothuria leucospilota

Opsonins, a group of serum proteins present in vertebrates, are known to bind to the surface of many kinds of pathogens and tag them as targets for phagocytosis. Opsonin-like molecules have been suggested to exist in several invertebrates. In this paper, we report the discovery of a new opsonin-like molecule in the coelomic fluid of the sea cucumber Holothuria leucospilota. Opsonic activity in cell-free coelomic fluid was characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, two-dimensional electrophoresis and gel filtration. Phagocytic uptake of yeast cells and latex beads by amoebocytes was quantified by microscopic analysis. It was found that a 20 kdalton protein bound to and enhanced phagocytosis of heat-killed yeast cells. The results of Western blot analysis suggest that both amoebocytes and spherule cells produce this opsonin-like molecule. Received: 22 October 1998 / Accepted: 4 March 2000     

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.     

Diel burying by the tropical sea cucumber Holothuria scabra: effects of environmental stimuli, handling and ontogeny

Understanding concealment behaviour of marine animals is vital for population surveys and captive-release programmes. The commercially valuable sea cucumber Holothuria scabra Jaeger 1833 (Holothuroidea) can display a diel burying cycle, but is it widely predictable? Circadian burying of captive H. scabra juveniles, and both juveniles and adults in the wild, was examined in New Caledonia. Groups of ten cultured juveniles in mesh chambers in a tank were monitored for 24 h. Small juveniles (1–5 g) displayed an expected diel cycle of epibenthic foraging in the afternoon and night then burial in sediments in the morning. Burial was related significantly to both light and temperature in combination. Similar groups of juveniles were handled once or three times a day for 1 week then frequency of emergence during another week was compared to unhandled controls. Handling stress, whether occasional or frequent, significantly suppressed the frequency of their afternoon emergence from sediments for 4 days. In a coastal seagrass bed, burial and emergence of H. scabra were monitored during days of opposing tidal cycles in three seasons. Adults seldom buried during the day except in the cool season. At that site, most small hatchery-produced H. scabra juveniles were buried during most of the day, while larger juveniles showed little diurnal burying. This study underscores that the circadian behaviours of marine animals can exhibit substantial spatial variation, may be absent at certain sites or seasons, and can be mediated by a complexity of factors that vary over short timescales.     

Effects of mercury graded doses on redox status,metallothionein levels and genotoxicity in the intestine of sea cucumber Holothuria forskali

Mercury (Hg) is a ubiquitous and a major environmental metal pollutant in the aquatic ecosystem. The present study was performed to evaluate the effect of mercury graded doses exposure on oxidative stress, redox status, metallothionein levels and genotoxicity in the intestine of sea cucumber Holothuria forskali. Specimens were exposed for 96?h to three concentrations of Hg (40, 80 and 160?µg/L). Exposure of H. forskali to Hg promoted oxidative stress with an increase in malondialdehyde (MDA), protein carbonyl (PCO) and advanced oxidation protein products (AOPP) levels. An increase of glutathione (GSH), vitamin C (ViteC) and non-protein thiols (NPSH) contents was also observed. Additionally, antioxidant activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) increased especially with the highest doses of Hg indicating a great defense of the antioxidant system. Our investigation revealed an increase in total Metallothionein (MT) content which was more pronounced at the sharpest concentrations of Hg. A significant decline of AChE activity was also observed. In the intestine of Hg-treated H. forskali genotoxicity was confirmed by DNA degradation.     

Reproduction of the commercial sea cucumber Holothuria whitmaei [Holothuroidea: Aspidochirotida] in the Indian and Pacific Ocean regions of Australia

Seasonal trends in the gonad index (GI) of two widely separated populations of black teatfish, Holothuria whitmaei (formerly included in Holothuria nobilis), were investigated between Pacific (Great Barrier Reef) and Indian Ocean (Ningaloo Reef) coral reefs of Australia. Reproductive activity followed a similar annual trend, with the GI of both populations peaking typically between April and June. Macroscopic and histological analysis of Ningaloo Reef specimens revealed that large germinal tubules, positioned centrally on the gonad basis, progressed through four maturity stages: growing (II); mature (III); partly spawned (IV); and spent (V). Growing tubules dominated the central region of the gonad basis between January and March, followed by an increase in the number of mature tubules throughout the GI peak (April–June). The progressive appearance of partly spawned and spent tubules between June and October suggests that spawning in H. whitmaei continues intermittently over an extended period throughout the austral winter. The examination of the gonad structure of sexually mature male and female specimens identified five tubule size classes in total (C₁ to C₅), each of differing physical and gametogenic status. In females, smaller C₁ tubules located at the anterior edge of the gonad basis contained pre- to early vitellogenic oocytes. Larger C₂ and C₃ tubule cohorts, positioned centrally on the gonad basis, contained mid- to late-stage vitellogenic oocytes. Smaller C₄ and C₅ tubules, located at the posterior edge, contained only relict oocytes. Similar physical and gametogenic differences were apparent between tubule cohorts in male specimens. We propose that these results, together with evidence of incomplete gonad resorption over the austral summer, indicate that gonad development in H. whitmaei conforms to the predictions of the Tubule Recruitment Model (TRM). The TRM is reported rarely among tropical aspidochirotes, and results presented here (1) provide the first direct evidence of this model in H. whitmaei, and (2) confirm that this species is one of the few winter-spawning tropical invertebrates.     

Role of secondary metabolites and pigments in the epidermal tissues, ripe ovaries, viscera, gut contents and diet of the sea cucumber Holothuria atra

The epidermal tissues, ovaries, viscera, gut contents, and the diet of Holothuria atra (Jaeger), collected from the Great Barrier Reef (GBR), Australia, just prior to spawning, contained carotenoid pigments, mycosporine-like amino acids (MAAs) and three other metabolites in varying proportions. Mycosporine-glycine (mycosporine-gly), palythine, asterina-330, shinorine, mycosporine-2-gly, porphyra-334, and palythinol were present in the epidermal tissues. Mycosporine-gly was the major MAA in the epidermal tissues, gut contents and the diet, and the only MAA present in the ripe ovaries and the viscera. Gadusol, a metabolite with antioxidant properties, which co-occurs with MAAs, and in certain instances with trace amounts of 6-deoxygadusol in unfertilised and fertilised eggs and developing larvae of some marine invertebrates and vertebrates, was absent from H. atra. However, 6-deoxygadusol, with similar physical and chemical properties to gadusol, and the proposed biosynthetic precursor of MAAs, was present, but only in the ovaries. This is first report of the presence of this proposed biogenetic precursor of MAAs in significant amounts in a marine invertebrate. Approximately 90% of the total carotenoids of the epidermal tissues, ovaries and viscera of H. atra are highly oxidised, the main component being astaxanthin followed by canthaxanthin. These were the major carotenoids present in the ovaries and the total carotenoid content was highest in the ovaries. β-carotene, a common egg carotenoid, was present in trace quantities in the ovaries but, along with the xanthophylls lutein and zeaxanthin, it occurred in significant amounts in the gut contents and the diet of H. atra. Carotenoid patterns in the epidermal tissues and viscera were strikingly similar, containing β-carotene, canthaxanthin, astaxanthin, zeaxanthin, lutein, phoenicoxanthin and echinenone. The alkaloids homarine and isomeric trigonelline, (structurally unrelated to MAAs) and 6-deoxygadusol were present only in the ovaries and the viscera. It is suggested that these metabolites and pigments are either involved with photoprotection or reproduction, or associated with both processes. The origin, biogenetic relationships and the roles of these metabolites and pigments in H. atra are discussed. Received: 21 November 1997 / Accepted: 14 September 1998     

Feeding factors for the sea anemone Anthopleura midorii

The carnivorous feeding behavior of the sea anemone Anthopleura midorii was found to be a sequence of successive feeding motions, which consisted of several separate actions: (1) tentaculation on any solid matter; (2) retention of prey by tentacles; (3) mouth opening; (4) ingestion of food; (5) digestion of food; (6) extrusion of indigestible waste material. Hot water extract of a natural prey induced the entire feeding sequence when extract was given in the form of an artificial food by mixing with -potato starch and water. The fractionated extract induced only a limited feeding action. Several amino acids induced feeding. Alanine, glycine or histidine stimulated the retention of artificial food by tentacles, and proline evoked mouth opening. Cysteine or reduced glutathione (GSH) induced food ingestion. Unnatural food stuff such as -potato starch was ingestible, if an appropriate amount of a chemical substance was added.     

The reproductive biology of a new species of sea cucumber, Holothuria (Mertensiothuria) arenacava in a Kenyan marine protected area: the possible role of light and temperature on gametogenesis and spawning

The sea cucumber Holothuria arenacava was discovered in the Mombasa marine reserve in 1997 and described by Samyn et al. (2001). The reproductive biology of this holothurian was investigated in order to (1) characterize the reproductive pattern, (2) examine the relationship among environmental parameters including temperature, light and lunar period, and (3) examine the relationship between the reproductive pattern and feeding of this new species. The gonad index method and microscopic examination of gonads was used to analyze samples collected for a period of 13 months. H. arenacava displayed an annual reproductive cycle with gametogenesis commencing in July during the south-east monsoons, when temperature and light intensity are lowest along the Kenyan coast. Gonad growth peaked in February–March at the end of the north-east monsoons when temperatures and light reach their annual maxima along the Kenyan coast. The higher correlation between light intensity and gonad growth (r=93) than temperature (r=0.71), coupled with the fact that temperatures continued to drop for a month after gametogenesis had already commenced, suggests that light intensity and not temperature is the cue for the onset of gametogenesis in this species. Spawning was synchronized between females and males and occurred during a short period between March and May (inter-monsoonal period) when both temperature and light intensity decrease along the Kenyan coast. Male and female gonad indices showed significant variation with lunar day and no lunar periodicity was observed in this sea cucumber. The sex ratio of the population of H. arenacava was skewed towards significantly more females than males, and females were significantly larger and had larger gonads and gonad indices than males. These life history strategies including spawning during a short discrete period, more and larger females that have larger gonads (i.e., typically more fecund), and spawning just prior to the peak in phytoplankton concentrations, a time that is probably more favorable for larval development, may serve to increase the reproductive success of this sea cucumber.     

Factors controlling spatial distribution of the sea cucumber Psolus chitonoides: Settling and post-settling behavior

Substratum selection experiments were carried out in situ with larvae of the holothurian Psolus chitonoides Clark in the rocky subtidal of the San Juan Islands, Washington. The experiments indicated, in agreement with laboratory findings, that pentactula larvae settle gregariously, either on the adult, or (when the bivium is obscured by epizooites) by clustering around the margin of the adult sole. Late larvae and early juveniles are strongly photonegative. Following metamorphosis, juveniles migrate from the adults into nearby shaded habitats, where they may take up residence indefinitely. Although adults retain the capacity to move, they relocate only when their living conditions become unsuitable; in most circumstances they are effectively sessile. Small-scale spatial pattern of the adults, which was documented quantitatively on subtidal rock walls, boulders and cobbles, correlates well with microtopographical features. The aggregation observed in the field is primarily due to post-settling migration rather than larval substratum choice at settlement.     

Feeding and nutritional ecology of the sea urchin strongylocentrotus drobachiensis in Maine,USA

The sea urchin strongylocentrotus drobachiensis exhibited a high degree of food selectivity, whether foods were presented singly or in combination. Foods ranked from most to least preferred were, in summer, Laminaria longicruris, Chondrus crispus, Corallina officinalis, Ascophyllum nodosum, and Agarum cribrosum, whereas in winter A. nodosum and A. cribrosum exchanged ranks. Food preference was not correlated with caloric content but, because of higher feeding rates on preferred foods, caloric intake was positively correlated with preference. Similarly, food absorption rankings were not correlated with food preference, with the exception of gravimetric efficiencies in winter. However, the absorption of L. longicruris, the most preferred alga, was highest in all measurements. Growth and reproductive development of S. drobachiensis on single species diets were positively correlated with food preference. Highest values occurred with animals fed L. longicruris. Urchins transferred from non-preferred to preferred diets showed increased somatic and reproductive growth compared to control animals on the original, non-preferred diets. Conversely, when transferred from preferred to non-preferred diets, urchins showed reduced growth compared to controls. No combination diet tested supported significantly better gonadal growth than L. longicruris: 25% C. crispus supported slightly better somatic growth than L. longicruris alone. These data support the view that S. drobachiensis has evolved a feeding strategy resulting in the maximization of growth and reproduction.     

Feeding of laboratory-reared larvae of the sea bream Archosargus rhomboidalis (Sparidae)

Feeding by larvae of the sea bream Archosargus rhomboidalis (Linnaeus) was investigated from late September, 1972 to early May, 1973 using laboratory-reared larvae. Fertilized eggs were collected from plankton tows in Biscayne Bay, and the larvae were reared on zooplankton also collected in plankton nets. Techniques were developed to estimate feeding rate, food selection, gross growth efficiency, and daily ration. Daily estimates of these were obtained through 16 days after hatching at rearing temperatures of 23°, 26°, and 29°C. Feeding rate increased exponentially as the larvae grew, and increased as temperature was raised. At 23°C larvae began feeding on Day 3, at 26° and 29°C larvae began feeding on Day 2. Feeding rates at initiation of feeding and on Day 16 were, respectively: 23°C, 7.16 food organisms per larva per hour (flh) and 53.78 flh; 26°C, 7.90 flh and 168.80 flh; 29°C, 17.62 flh and 142.07 flh. Sea bream larvae selected food organisms by size. At initiation of feeding they selected organisms less than 100 m in width. As larvae grew they selected larger organisms and rejected smaller ones. The major food (more than85% of the organisms ingested) was copepod nauplii, copepodites, and copepod adults. Minor food items were barnacle nauplii, tintinnids, invertebrate eggs, and polychaete larvae. Mean values for gross growth efficiency of sea bream larvae ranged from 30.6% at 23°C to 23.9% at 29°C. Mean values for daily ration, expressed as a percentage of larval weight, ranged from 84% at 23°C to 151% at 29°C and tended to decline as the larvae grew.This paper is a contribution from the Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, Florida, USA     

Feeding behavior and acquisition of zooxanthellae by planula larvae of the sea anemone Anthopleura elegantissima

Symbiotic associations between cnidarians and photosynthetic dinoflagellates (i.e., zooxanthellae) are common in the marine environment. Many symbiotic cnidarians produce offspring that are initially nonsymbiotic. These new hosts must acquire symbiotic algae from environmental sources. We examined zooxanthella acquisition by laboratory-reared planula larvae of the temperate sea anemone Anthopleura elegantissima. Larvae ingested zooxanthellae while they were feeding. However, the signal that prompted larval feeding behavior did not originate from the symbiotic algae; the addition of algal cells to larval cultures never elicited a feeding response. In contrast, the addition of macerated animal tissue from several sources invariably generated a strong feeding response, which resulted in the larvae indiscriminately ingesting any particulate matter that was present, including zooxanthellae or other unicellular algae. Ingested zooxanthellae were incorporated into endodermal cells, where they remained undigested, while all other ingested material was digested or expelled within 24 h. Our results provide evidence that one source of zooxanthellae likely to serve as a route of infection in the natural environment is zooxanthella-laden mucus egested by anemones. This egested material fulfilled both of the criteria necessary for successful infection: it prompted larvae to begin feeding and provided an abundant supply of zooxanthellae that were ingested and taken up into endodermal cells of the new host.     

Using demography and movement behavior to predict range expansion of the southern sea otter

In addition to forecasting population growth, basic demographic data combined with movement data provide a means for predicting rates of range expansion. Quantitative models of range expansion have rarely been applied to large vertebrates, although such tools could be useful for restoration and management of many threatened but recovering populations. Using the southern sea otter (Enhydra lutris nereis) as a case study, we utilized integro-difference equations in combination with a stage-structured projection matrix that incorporated spatial variation in dispersal and demography to make forecasts of population recovery and range recolonization. In addition to these basic predictions, we emphasize how to make these modeling predictions useful in a management context through the inclusion of parameter uncertainty and sensitivity analysis. Our models resulted in hind-cast (1989-2003) predictions of net population growth and range expansion that closely matched observed patterns. We next made projections of future range expansion and population growth, incorporating uncertainty in all model parameters, and explored the sensitivity of model predictions to variation in spatially explicit survival and dispersal rates. The predicted rate of southward range expansion (median = 5.2 km/yr) was sensitive to both dispersal and survival rates; elasticity analysis indicated that changes in adult survival would have the greatest potential effect on the rate of range expansion, while perturbation analysis showed that variation in subadult dispersal contributed most to variance in model predictions. Variation in survival and dispersal of females at the south end of the range contributed most of the variance in predicted southward range expansion. Our approach provides guidance for the acquisition of further data and a means of forecasting the consequence of specific management actions. Similar methods could aid in the management of other recovering populations.     

Feeding ecology of the loggerhead sea turtle Caretta caretta in the Northwestern Gulf of Mexico

Digestive tract contents collected from carcasses of 82 loggerhead sea turles (Caretta caretta) found on the south Texas coast from 1986 through 1988 were examined. Benthic invertebrates were the predominant prey. Sea pens (Virgularia presbytes), crabs, and mollusks accounted for 94% of the dry weight of the digestive tract samples. Temporal changes in the percent occurrence and percent dry weight of sea pens, crabs, and mollusks in the digestive tract samples were significant. Loggerheads fed primarily on sea pens during spring, then primarily on crabs during summer and fall. The increase of crabs in the loggerhead diet paralleled the annual increase in the abundance of crabs in the Gulf of Mexico. Sea pens were located nearshore at depths of 6 to 12 m and had a disjunct distribution. Present address: Texas A&M University, Department of Biology, College Station, Texas 77843-3258, USA     

Changes of movement patterns from early dispersal to settlement

Moving and spatial learning are two intertwined processes: (a) changes in movement behavior determine the learning of the spatial environment, and (b) information plays a crucial role in several animal decision-making processes like movement decisions. A useful way to explore the interactions between movement decisions and learning of the spatial environment is by comparing individual behaviors during the different phases of natal dispersal (when individuals move across more or less unknown habitats) with movements and choices of breeders (who repeatedly move within fixed home ranges), that is, by comparing behaviors between individuals who are still acquiring information vs. individuals with a more complete knowledge of their surroundings. When analyzing movement patterns of eagle owls, Bubo bubo, belonging to three status classes (floaters wandering across unknown environments, floaters already settled in temporary settlement areas, and territory owners with a well-established home range), we found that: (1) wandering individuals move faster than when established in a more stable or fixed settlement area, traveling larger and straighter paths with longer move steps; and (2) when floaters settle in a permanent area, then they show movement behavior similar to territory owners. Thus, movement patterns show a transition from exploratory strategies, when animals have incomplete environmental information, to a more familiar way to exploit their activity areas as they get to know the environment better.     

Distribution patterns of plants explained by human movement behaviour

Distribution patterns of plants are affected by human activities such as creation, destruction or modification of habitats. However, another important question is to what extent humans shape plant distributions by acting as dispersal vectors. In order to answer this question we developed a simulation model for the spread of plant species between human settlements. This was done on the basis of extensive sociological and ecological data on a regional scale. With regard to the sociological data, human movement behaviour defined the amount of exchange between the settlements. Gardening types represented the potential habitat in our model. The ecological data was derived from a vegetation survey carried out in 2003, which was a repeat of a survey between 1974 and 1981 along the same transects. From these surveys, we studied the distributions of 13 species in 67 settlements. In our model, the earlier survey provided the data for the initial distribution. The simulated pattern was consequently compared with the distribution pattern in 2003. In the model, dispersal kernels based on patterns of human movement between settlements led to a better match with the distribution patterns than a null model simulating pure distance dependent dispersal for all species. This was statistically significant for seven of the thirteen species. A striking result was that alien species seem to benefit more from human dispersal than native species. We emphasize the importance of the sociological data on human movement behaviour in parameterizing our regional scale model. This study provides quantitative evidence on the impact of human movement behaviour on the distribution of plant species in suburban areas.     

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.