The reproductive response of the sea urchins Paracentrotus lividus (G.) and Psammechinus miliaris (L.) to a hyperproteinated macrophytic diet

The sea urchins Paracentrotus lividus and Psammechinus miliaris are submitted to the same environmental conditions in the Bay of Brest. The relationship between seasonal changes in food source quality and their gonad production was investigated in reproducing experimentally these conditions. In a first stage two macroalgae (Palmaria palmata and Laminaria digitata) were tested. P. miliaris showed a stronger preference for P. palmata and over a year-long experiment both urchins progressively preferred P. palmata. Seasonal variations in the chemical composition of P. palmaria were observed in the Bay of Brest: total carbohydrates were important and the relative maximum (about 50%) was reached between February and August; the lipid level was low and had a relative maximum of about 1% in June and August. Total protein in P. palmaria was high compared to other seaweeds: the maximum value (25%) was observed in June, which was probably due to the maintenance of nitrogen nutrient in the bay.In the second stage of the study, seasonal changes in biochemical components of ingestion and absorption of the two sea urchins were followed in the laboratory using a monospecific diet of P. palmaria. The patterns of total carbohydrates and lipid absorption were very similar for both sea urchin species. Carbohydrates were absorbed strongly and uniformly, year round. Lipid absorption mimicked the lipid nutrient pattern in the food source. Only changes in protein absorption varied slightly between the two urchin species. Protein absorption was maximal for both species in February and June, but the quantity of absorbed protein was significantly higher in P. miliaris than in P. lividus during February. This increase was concomitant with protein storage in the sea urchin gonads, which peaked in February for P. miliaris and in June for P. lividus. P. lividus had a higher gonad production efficiency, based on gonad yield. The comparison between in situ data and the experimental results suggests that an algal diet more nitrogenous than the in situ algal food source would benefit the herbivorous P. lividus, rather than the more omnivorous species P. miliaris. Although P. milaris has been described as a species with large gonad production potential, P. lividus appears to be a more suitable species for echiniculture conditions.     

Developmental aberrations associated with twinning in laboratory-reared sea urchins

Twinned sea urchins of Arbacia punctulata were reared in the laboratory from single first cleavage blastomeres through metamorphosis to reproductive adults. A pair of metamorphosed urchins, reared in the same container, with adequate food, typically differed in size and gross morphology. Aberrant plate variations of the normal apical system arrangement for the species were expressed by a significantly greater number of twinned urchins than controls derived from whole zygotes. The influence of early embryogenesis on the development of adult features is discussed.     

Toxic Diatom Aldehydes Affect Defence Gene Networks in Sea Urchins

Marine organisms possess a series of cellular strategies to counteract the negative effects of toxic compounds, including the massive reorganization of gene expression networks. Here we report the modulated dose-dependent response of activated genes by diatom polyunsaturated aldehydes (PUAs) in the sea urchin Paracentrotus lividus. PUAs are secondary metabolites deriving from the oxidation of fatty acids, inducing deleterious effects on the reproduction and development of planktonic and benthic organisms that feed on these unicellular algae and with anti-cancer activity. Our previous results showed that PUAs target several genes, implicated in different functional processes in this sea urchin. Using interactomic Ingenuity Pathway Analysis we now show that the genes targeted by PUAs are correlated with four HUB genes, NF-κB, p53, δ-2-catenin and HIF1A, which have not been previously reported for P. lividus. We propose a working model describing hypothetical pathways potentially involved in toxic aldehyde stress response in sea urchins. This represents the first report on gene networks affected by PUAs, opening new perspectives in understanding the cellular mechanisms underlying the response of benthic organisms to diatom exposure.     

Loss of sex-allocation plasticity in the predatory mite Metaseiulus occidentalis and possible triggering cues

In two recent studies, we showed that females from a long-established Metaseiulus (Typhlodromus or Galendromus) occidentalis laboratory colony reared under crowded conditions for approximately 27 years produced equal numbers of male and female progeny. This contrasts with our previous results that showed the females from the same colony produced progeny with a female-biased sex ratio. These discrepant findings raised the possibility that dense rearing conditions can affect the sex-allocation plasticity in this important biological control agent. In the current study, we show that females from a newly collected M. occidentalis colony initially were capable of producing progeny with a female-biased sex ratio, but lost this ability over a 12-month period (43 generations) when reared under dense rearing conditions and provided with damaged prey. We demonstrate that females from this newly collected colony initially could adjust offspring sex ratio in response to both conspecific cues and cues produced by their damaged prey. The results suggest that dense rearing conditions, which are often used in commercial rearing facilities, may have detrimental effects on the reproductive fitness of M. occidentalis and, possibly, other predatory mites.     

In situ cryopreservation of human embryonic stem cells in gas-permeable membrane culture cassettes for high post-thaw yield and good manufacturing practice

Current strategies for marine pollution monitoring are based on the integration of chemical and biological techniques. The sea urchin embryo-larval bioassays are among the biological methods most widely used worldwide. Cryopreservation of early embryos of sea urchins could provide a useful tool to overcome one of the main limitations of such bioassays, the availability of high quality biological material all year round. The present study aimed to determine the suitability of several permeant (dimethyl sulfoxide, Me₂SO; propylene glycol, PG; and ethylene glycol, EG) and non-permeant (trehalose, TRE; polyvinylpyrrolidone, PVP) cryoprotectant agents (CPAs) and their combination, for the cryopreservation of eggs and embryos of the sea urchin Paracentrotus lividus. On the basis of the CPAs toxicity, PG and EG, in combination with PVP, seem to be most suitable for the cryopreservation of P. lividus eggs and embryos. Several freezing procedures were also assayed. The most successful freezing regime consisted on cooling from 4 to −12 °C at 1 °C/min, holding for 2 min for seeding, cooling to −20 °C at 0.5 °C/min, and then cooling to −35 °C at 1 °C/min. Maximum normal larvae percentages of 41.5% and 68.5%, and maximum larval growth values of 42.9% and 60.5%, were obtained for frozen fertilized eggs and frozen blastulae, respectively.     

Consumers of sea urchins,<Emphasis Type="Italic"> Paracentrotus lividus</Emphasis> and<Emphasis Type="Italic"> Arbacia lixula</Emphasis>, in shallow Mediterranean rocky reefs

Underwater observations on fish and asteroid consumers (i.e. predators and scavengers) of sea urchins, Paracentrotus lividus and Arbacia lixula, were carried out at several locations in shallow Mediterranean rocky reefs. Observations conducted in the marine reserve of Torre Guaceto (Adriatic Sea) revealed that sparid fishes, Diplodus sargus and D. vulgaris, are the main fish predators of small (<1 cm in test diameter) and medium (1–4 cm) sea urchins, whereas the labrids Coris julis and Thalassoma pavo preyed only upon small sea urchins. Large D. sargus were able to prey upon small and medium, and occasionally large (>4 cm) sea urchins, whereas medium and small Diplodus preyed mainly upon small sea urchins. The number of sea urchins preyed upon by fishes was negatively related to sea urchin size for both species. P. lividus appeared to be subject to higher predation levels than A. lixula. The scavenger guild comprised 11 fish species, with D. sargus, D. vulgaris, Coris julis and Chromis chromis accounting for about 80% of scavenger fishes. Observations performed at several locations in the Mediterranean on the predatory asteroid Marthasterias glacialis revealed that only 3% of the detected individuals were preying upon sea urchins. Due to the importance of sea urchins for assemblage structure and functioning of Mediterranean rocky reef ecosystems, these results may have also important implications for management of fishing activities.Communicated by H.-D. Franke     

Lissarca notorcadensis (Bivalvia: Philobryidae) living on Notocidaris sp. (Echinoidea: Cidaridae): Population dynamics in limited space

Summary Population dynamics of the epizoic bivalve Lissarca notorcadensis living on spines of cidaroid sea urchins in the Weddell Sea were investigated. Total production (somatic & gonad) of the suspension feeding bivalve ranged between 16.5 and 487.4 mg AFDM y^–1 per sea urchin. Annual sedimentation rates are not sufficient to maintain the production of the Lissarca sub-populations carried by the sea urchins, and resuspension of organic matter is most likely to be an important food source. The ratio of the number of freshly settled juveniles to the number of embryos brooded is between 0.054 and 0.207 and seems negatively related to the biomass already present, indicating intraspecific competition for space. Interspecific competition for space is caused by the strong preference of L. notorcadensis as well as other epizoa (colonial anthozoans and bryozoans) for the spines located on the aboral hemispere of the sea urchins.AWI Publication No. 572     

Environmentally relevant cadmium concentrations affect development and induce apoptosis of <Emphasis Type="Italic">Paracentrotus lividus</Emphasis> larvae cultured in vitro

Sea urchin embryos and larvae represent suitable model systems on where to investigate the effects of heavy metals on development and cell viability. Here, we tested the toxic effects of low (10⁻¹² M), medium (10⁻⁹ M), and high (10⁻⁶ M) cadmium chloride concentrations, mimicking unpolluted, moderately and highly polluted seawaters, respectively, on Paracentrotus lividus sea urchins offspring. Larvae were continuously treated from fertilization and inspected at time intervals comprised between 10 and 30 days of development. Delays and/or morphological abnormalities were firstly evident in larvae treated for 15 days with high cadmium (10⁻⁶ M) and for 25 days with medium cadmium (10⁻⁹ M). Major defects consisted in the reduction and lack of arms and skeleton elongation. No obvious differences with respect to controls were observed in embryos/larvae exposed to low cadmium (10⁻¹² M), even after 30 days of exposure. Using in situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay on larvae whole mounts, we detected apoptosis after 10 days of treatment with 10⁻⁶ and 10⁻⁹ M CdCl_2, when no morphological abnormalities were recognizable yet. Supernumerary apoptotic cells were found in arm buds, ciliary bands, and apex. In conclusion, echinoderm embryos and larvae represent candidates of choice for the study of stress and defense mechanisms activated by cadmium exposure.     

Reproductive biology of the sea anemone shrimp Periclimenes rathbunae (Caridea,Palaemonidae, Pontoniinae), from the Caribbean coast of Costa Rica

Caridean shrimps are a highly diverse group and many species form symbiotic relationships with different marine invertebrates. Periclimenes rathbunae is a brightly colored shrimp that lives predominantly in association with sea anemones. Information about the reproductive ecology of the species is scarce. Therefore, we collected 70 ovigerous females inhabiting the sun sea anemone Stichodactyla helianthus in coral reefs from the southern Caribbean coast of Costa Rica. Females produced on average 289 ± 120 embryos. The volume of recently-produced embryos was on average 0.038 mm³, and embryo volume increased by 192% during the incubation period. The average embryo mortality during embryogenesis was 24%. The reproductive output was 0.24 ± 0.094, considerably higher than in many other pontoniine shrimps. Females carrying embryos close to hatching showed fully developed ovaries, suggesting consecutive spawning. We assume that the sheltered habitat, living on sea anemones, allows Periclimenes rathbunae to allocate more energy in embryo production than most other free-living caridean shrimps. This is the first record of Periclimenes rathbunae for Costa Rica.     

Variation among females in egg lipid content and developmental success of echinoderms from McMurdo Sound,Antarctica

Egg lipid and protein content of different females of Antarctic echinoderms in McMurdo Sound, Antarctica, were measured to assess variation among females and developmental success. Egg triacylglycerol content of the Antarctic sea urchin Sterechinus neumayeri, when less than 70 ng, correlated with embryos that failed to develop past the 4-day-old blastula stage. In contrast asteroids (Odontaster meridionalis, O. validus, Acodontaster hodgsoni) all produced eggs that developed normally, even with variable egg lipid content. This difference might be related to dietary sources for more herbivorous sea urchins compared to more omnivorous and predatory asteroids. Low egg lipid content, with resulting poor embryonic survivorship, suggests that herbivorous sea urchins may be under unusual levels of nutritional stress in McMurdo Sound during the time frame studied (2004–2005). This nutritional stress might be related to the presence of large icebergs, known to have reduced primary production in the Ross Sea area.     

Morphologic der seeigelzähne II. Die gekielten Zähne der echinacea (Echinodermata,echinoidea)

Structure and growth of a keeled tooth are discussed with Paracentrotus lividus as the main example. The microscopic structure of the tooth skeletons of other sea urchins was compared, considering at least one member of each family with the exception of Saleniidae. Attention is called to the necessary revision of the system of sea urchins.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

遮蔽行为对海刺猬摄食、生长和性腺性状的影响

海胆遮蔽行为对于海胆的生长和发育的影响目前还不清楚,在实验室条件下,研究了遮蔽行为对海刺猬(Glyptocidariscrenularis)摄食、生长和性腺性状的影响。实验设两个对照组和一个处理组:对照组为无任何遮蔽条件的空白组和以堆积砖块创造的黑暗空间来满足海胆的掩蔽行为的掩蔽组,遮蔽组则以菲律宾蛤仔(Ruditapes philippinarum)的贝壳作为遮蔽物来满足海胆的遮蔽行为。结果显示:遮蔽组的海胆周摄食量要显著大于掩蔽组(P<0.05),而空白组与遮蔽组和掩蔽组之间的差异不显著(P>0.05);各组海胆的存活率没有显著差异(P>0.05);空白组和遮蔽组海胆的壳径、壳高、体重要显著大于掩蔽组(P<0.05),而空白组和遮蔽组之间没有显著性的差异(P>0.05);空白组和遮蔽组的性腺湿重、性腺干重、性腺指数、壳湿重、壳干重、亚氏提灯湿重、亚氏提灯干重要显著大于掩蔽组(P<0.05),而空白组和遮蔽组之间没有显著性的差异(P>0.05);空白组的提灯指数要显著小于遮蔽组和掩蔽组(P<0.05),而遮蔽组和掩蔽组之间没有显著的差异(P>0.05)。结果证明,遮蔽和掩蔽行为对海刺猬生长及发育的影响是有差别的,掩蔽行为对海刺猬的生长与发育造成了负面影响,而遮蔽行为在一定的生活环境条件下不会对其生长造成影响。研究表明,海刺猬的遮蔽行为反映了其对外界不利环境条件的一种适应性,且这种行为对其生长和发育的影响与掩蔽行为有所不同。至于海刺猬遮蔽行为的进化选择压力,目前的研究还不足以下定论,需要进一步深入的研究。     

Effects of oil pollution on the development of sex cells in sea urchins

The sea urchinStrongylocentrotus nudus is highly sensitive to oil pollution. Experiments were performed in winter, spring and summer over periods of 15 to 45 days. Experimental urchins were kept in water with hydrocarbon concentrations of 10 to 30 mg l⁻¹, and control urchins in pure sea water. Thermal stimulation by Evdokimov's method was applied to obtain mature sexual products during winter and spring tests. Summer investigations were conducted at temperatures of 17 to 18 °C. The gonads were studied histologically and morphometrically, and the sexual cells obtained were analyzed at the embryological level. No histological and morphometrical differences were recorded between sexual cells of controls and experimentals. However, marked hydrocarbon effects were observed in the embryonic development of artificially fertilized cells from experimental urchins. Control embryos developed normally. Embryogenesis of artificially fertilized gametes from control females and experimental males, and vice versa, was found to be distinctly abnormal. Many abnormalities were identified at the first cleavage stage, as well as in blastula, gastrula and pluteus. Fertilization of experimental eggs with experimental sperm resulted in serious disturbances of embryos, followed by the development of non-viable larvae. On the whole, embryogenesis of sexual cells from experimental urchins was characterized by prominent delay, asynchronism and presence of abnormal non-viable larvae. Consequently, long-term effects of sublethal hydrocarbon concentrations resulted in the formation of defective sex cells and high larval mortality.     

Intracytoplasmic Sperm Injection Using DNA-Fragmented Sperm in Mice Negatively Affects Embryo-Derived Embryonic Stem Cells,Reduces the Fertility of Male Offspring and Induces Heritable Changes in Epialleles

Intracytoplasmic sperm injection (ICSI) in mice using DNA-fragmented sperm (DFS) has been linked to an increased risk of genetic and epigenetic abnormalities both in embryos and offspring. This study examines: whether embryonic stem cells (ESCs) derived from DFS-ICSI embryos reflect the abnormalities observed in the DFS-ICSI progeny; the effect of DFS-ICSI on male fertility; and whether DFS-ICSI induces epigenetic changes that lead to a modified heritable phenotype. DFS-ICSI-produced embryos showed a low potential to generate ESC lines. However, these lines had normal karyotype accompanied by early gene expression alterations, though a normal expression pattern was observed after several passages. The fertility of males in the DFS-ICSI and control groups was compared by mating test. Sperm quantity, vaginal plug and pregnancy rates were significantly lower for the DFS-ICSI-produced males compared to in vivo-produced mice, while the number of females showing resorptions was higher. The epigenetic effects of DFS-ICSI were assessed by analyzing the phenotype rendered by the Axin1^Fu allele, a locus that is highly sensitive to epigenetic perturbations. Oocytes were injected with spermatozoa from Axin1^{Fu /+} mice and the DFS-ICSI-generated embryos were transferred to females. A significantly higher proportion of pups expressed the active kinky-tail epiallele in the DFS-ICSI group than the controls. In conclusion: 1) ESCs cannot be used as a model of DFS-ICSI; 2) DFS-ICSI reduces sperm production and fertility in the male progeny; and 3) DFS-ICSI affects the postnatal expression of a defined epigenetically sensitive allele and this modification may be inherited across generations.     

Ontogenetic Shifts in Patterns of Microhabitat Utilization in the Small-headed Clingfish, Apletodon Dentatus (Gobiesocidae)

The patterns of microhabitat utilization by the clingfish Apletodon dentatus were investigated, based on SCUBA diving surveys at the Arrábida Marine Park (Portugal). In all inspected microhabitats, this species was only found in algal turfs, sea urchins and boulders. The association of A. dentatus with sea urchins is here analysed for the first time. There was a differential utilization of the microhabitats, with small juveniles recruiting to algal turfs, intermediate individuals found in association with the sea urchins Paracentrotus lividus and Sphaerechinus granularis and larger fish occurring mainly in boulders. The depth distribution patterns are also analysed.     

Effects of NO-Synthase Inhibitors on Development of Sea Urchin Embryos

Effects of NO-synthase inhibitors N-nitro-L-arginine (L-NA) and its methylated ether (L-NAME) on embryonic development of sea urchins Paracentrotus lividus and Arbacia lixula were studied from the time of fertilization to the stage of transition to active nutrition (stage of the later pluteus 2). It has been shown that L-NAME (but not D-NAME) and L-NA (0.01–0.02 mM) produce a dose-dependent inhibition of growth of arms and apex of pluteus larvae, while differentiation of the intestine, coelom, pigment cells, and ciliated epithelium occurs without observable disturbances. A period of sensitivity to NO-synthase inhibitors was revealed; it coincided with the beginning of intensive spiculogenesis leading to elongation of arms and apex of the pluteus larva of the stage (prism 2—early pluteus 2). It is suggested that interaction of ectodermal cells with the primary mesenchime cells and extracellular matrix in morphogenetic processes providing formation of arms and apex of the pluteus larva can be modulated by NO in ontogenesis of sea urchins P. lividus and A. lixula.     

Chemical composition of spines and tests of sea urchins

The skeleton of spines and tests of the species of sea urchins Strongylocentrotus intermedius, Mesocentrotus nudus, Scaphechinus mirabilis, and Echinocardium cordatum from the Sea of Japan is composed of a spongy stereom, consisting of calcite with a high content of magnesium. It was found that the tests and spines of the skeletons of sea urchins are composed of calcium–organic composite materials inlaid with other metals: Mg, Fe, Zn, and Rb. In the four species of sea urchins studied, the strength and other mechanical properties of the tests and spines differ and depend on the chemical composition and structural organization of their components. It was shown that the content of volatile substances correlates with their fragility or elasticity. It is revealed that the chemical composition of the tests of two species of the spherical sea urchins S. intermedius and M. nudus indicates significant differences between these two species of sea urchins.