Induction of hatching by chemical signals secreted by the ovigerous female of an estuarine crab Sesarma haematocheir

Hatching of embryos in the estuarine crab Sesarma haematocheir is highly synchronized with nocturnal high tide and completes within 1 hr among all embryos incubated by the female. This highly synchronized hatching is induced by a "Hatching-Program Inducing Factor (HPIF)" released from the female. To further define the cues involved in synchronized hatching, experiments were designed to characterize this factor and to determine possible sites of release and temporal release patterns using strategies involving isolation of egg masses, perfusion, and ablation experiments on fully developed embryos that had not yet entered the hatching program. Embryo transplantations indicate that not only HPIF may be released from the branchial chamber, but that it is extraordinarily unstable, and loses activity within 15 min, which frustrates further attempts at characterization. Nevertheless, with regard to temporal release patterns, it was established that HPIF activity was detected during short periods over three consecutive nights prior to release of larvae. This activity did not explain the gated response of embryo release in this crab, which might correspond with circatidal larval release events in the field.     

A pair of rosette glands in the embryo and zoeal larva of an estuarine crab Sesarma haematocheir, and classification of the tegumental glands in the embryos of other crabs

A pair of rosette glands (one of the tegumental glands in crustaceans) is present at the root of the dorsal spine of the thorax in mature embryos of the estuarine crab Sesarma haematocheir. Each rosette gland is spherical, 45-50 microm in diameter. This gland consists of three types of cells: 18-20 secretory cells, one central cell, and one canal cell. The secretory cells are further classified into two types on the basis of the morphology of secretory granules. There are 17-19 a cells, and only one b cell per rosette gland. An a cell contains spherical secretory granules of 2-3 microm in diameter. The granules are filled with highly electron-dense materials near the nucleus but have lower electron-density near the central cell. The secretory granules contained in the b cell have an irregular shape and are 1-1.5 microm in diameter. The density of the materials in the granules is uniform throughout the cytoplasm. The secretory granules contained in both the a and b cells are produced by the rough endoplasmic reticulum. Materials in the granules are exocytotically discharged into the secretory apparatus inside the secretory cell, sent to the extracellular channels in the central cell, and secreted through the canal cell. The rosette gland can be distinguished from the epidermal cells 2 weeks after egg-laying and the gland matures just before hatching. Materials produced by this gland are secreted after hatching and secretion continues through five stages of zoeal larvae. These rosette glands were never found in the megalopal larva. Rosette glands are found in the embryos of Sesarma spp. and Uca spp. In other crabs, tegumental glands are also found at the same position as in the embryo of S. haematocheir, but the fine structure of their glands is largely different from that of the rosette gland. On the basis of the morphology of secretory cells (a-g cell types), the tegumental glands of a variety of crab embryos can be classified into four types, including rosette glands (type I-IV). The function of these tegumental glands is not yet known, but different types of the gland seem to reflect the phylogeny of the crabs rather than differences of habitat.     

Hatching of an estuarine crab, Sesarma haematocheir: from disappearance of the inner (E3) layer to rupture of the egg case

Hatching of decapod crustaceans is characterized by the sudden rupture of the egg case. This study focused on the following two issues regarding the hatching mechanism of the estuarine terrestrial crab Sesarma haematocheir: (1) dissolution of the egg case, and (2) the site where the egg case breaks. The egg case comprises three layers: the outer two (E1 and E2) layers and the inner (E3) thin layer (0.2 microm in thickness). The outer layers showed no morphological changes upon hatching, but the inner layer (E3) was markedly digested. The digestion of this layer would enable the embryo to absorb ambient water via reverse peristalsis of the intestine, resulting in an increase of the volume. The egg case always ruptured perpendicular to the longitudinal axis of the embryo. In addition, breakage of the egg case occurred at the dorsal thorax of the embryo. The three major organs positioned at this area were (1) a sharp projection (dorsal spine), (2) an assemblage of muscles, and (3) a pair of secretory glands, each of which was about 30 microm in diameter. The dorsal projection is soft before hatching, and it is clear that the egg case does not break with the posterior expansion of this projection. The rupture instead appears to be caused by the expansion of the muscles arranged perpendicular to the body axis. In addition, some (unknown) factor might weaken the egg case just before hatching. The secretory glands may be a kind of rosette gland, but the role that this gland plays at hatching is not known. As a duct comes out from the center and enters the dorsal projection, some active substance may be released at the tip of this projection. However, immunochemical studies are not consistent with this substance being an ovigerous hair stripping substance (OHSS).     

Entrainment of a semilunar rhythm by a simulated moonlight cycle in the terrestrial crab,Sesarma haematocheir

Summary Sesarma haematocheir is a species of terrestrial crabs inhabiting hillsides and paddy fields near the sea. Females show a semilunar rhythm of zoea-release coinciding with days of spring tides and in addition with the time of high water occurring at the nearby seacoast about dusk. Nature of environmental stimuli or zeitgebers that induce the semilunar rhythm of zoea-release was examined experimentally. In case that a population of males and females was kept under the condition of a 24-h light-dark cycle (LD 14:10) only, females showed a free-running semilunar rhythm of egg-laying and zoea-release synchronized from field conditions. On the other hand, a semilunar rhythm of egg-laying and zoea-release was entrained by the combination of the 24-h LD and a simulated 24.8-hr moonlight cycle of which the phase was shifted in relation to the natural lunar cycle. This result suggests that the 24.8-h moonlight cycle acts as a zeitgeber of a semilunar rhythm. The 12.4-h tidal cycle parallels with the 24.8-h moonlight cycle in the field. On the basis of the perception of a distinct phase relationship between the 24-h LD and the 24.8-h moonlight cycle, it is considered that crabs substitute the 24.8-h moonlight cycle for the 12.4-h cycle of tides as a zeitgeber to synchronize the phase of the semilunar rhythm with a tidal situation.     

中华绒螯蟹胚胎附着系统的结构和形成机制

利用透射电镜和扫描电镜技术研究了中华绒螯蟹(Eriocheir sinensis)胚胎附着系统的结构及形成机制。中华绒螯蟹受精卵附着在雌性腹肢内肢的携卵刚毛上。该附着系统由三个连续部分组成:卵膜、卵柄和被膜,后者覆盖在携卵刚毛的绒毛上。研究结果显示:中华绒螯蟹的成熟胚胎由三层明显的卵膜组成,即E1、E2和E3层,但胚胎附着系统的卵柄及被膜仅为外层(E1)。卵巢中成熟卵的卵膜仅由E1层组成,E1分为两个亚层(E1a′、E1b′)。胚胎附着系统的形成与雌蟹的行为、腹肢粘液腺分泌的粘液、卵膜的超微结构及各层的变化有关。受精卵刚从生殖孔中排出时,卵膜(E1a′、E1b′)并不能直接粘附在携卵绒毛上。产卵后不久,雌蟹腹肢粘液腺分泌粘液的量增多,E1a′、E1b′的结构发生变化,表现为边界模糊,卵膜出现很强的粘性。在产卵后约60min E1层又明显分为两个亚层(E1a、E1b),同时排卵后雌蟹腹部的携卵绒毛不断地运动,这种运动促使携卵绒毛外的被膜形成。随着E1层亚结构的变化,E2层也开始形成,当E1新的两个亚层出现时,部分区域的E1层与E2层发生分离,卵柄开始形成,并牢固地附着在携卵绒毛上。被膜、卵柄与卵膜最外层的结构相同,均由E1层构成[动物学报51(5):852-861,2005]。     

Structure, attachment properties, and ecological importance of the attachment system of English ivy (Hedera helix)

Root climbers such as English ivy (Hedera helix) rely on specialized adventitious roots for attachment, enabling the plants to climb on a wide range of natural and artificial substrates. Despite their importance for the climbing habit, the biomechanical properties of these specialized adventitious roots compared with standard roots and their performance in the attachment to different host species or inert substrates have not been studied. Here organs and tissues involved in the attachment are characterized and their significance in regard to a broader functional and ecological aspect is discussed. Depending on the substrate, the root clusters show different types of failure modes at various frequencies, demonstrating the close interaction between the climber and its substrates. With a Young's Modulus of 109.2 MPa, the attachment roots are relatively stiff for non-woody roots. The central cylinders of the attachment roots show a high tensile strength of 38 MPa and a very high extensibility of 34%. In host trees naturally co-distributed with English ivy, a 'balanced' occurrence of failure of the attachment system of the climber and the bark of the host is found, suggesting a co-evolution of climber and host. Maximum loads of root clusters normalized by the number of roots match those of individually tested attachment roots. In comparison with most subterranean roots the properties and structure of the attachment roots of English ivy show distinct differences. There exist similarities to the properties found for roots of Galium aparine, suggesting a trend in not fully self-supporting plants towards a higher extensibility.     

Some changes in biochemical composition with sex and size of the crab,Sesarma boulengeri calman

Variations in the biochemical composition (protein and cholesterol) have been studied in the hepatopancreas and thoracic muscle of the crab, Sesarma boulengeri as a function of sex and size. Second degree polynomial regression equations are used in predictive fashion to quantify the biochemical constituents in relation to sex and size. The effect of size is expressed as a power function of body weight. A good agreement is found between the experimental data and the theoretical values based on the polynomial.There is a negative correlation between the mass of crab and the reduced mass of cholesterol and protein of the tissues in both sexes, signifying higher metabolic and growth rates in younger crabs. The cholesterol and protein contents are higher in the male indicating more growth than in the female. However, there is tendency of the muscle protein to increase in large sized female crabs suggesting that bigger sized female are structurally better suited than males.     

Foraging behavior of an estuarine predator, the blue crab Callinectes sapidus in a patchy environment

To define general principles of predator‐prey dynamics in an estuarine subtidal environment, we manipulated predator density (the blue crab, Callinectes sapidus) and prey (the clam, Macoma balthica) patch distribution in large field enclosures in the Rhode River subestuary of the central Chesapeake Bay. The primary objectives were to determine whether predators forage in a way that maximizes prey consumption and to assess how their foraging success is affected by density of conspecifics. We developed a novel ultrasonic telemetry system to observe behavior of individual predators with unprecedented detail. Behavior of predators was more indicative of optimal than of opportunistic foraging. Predators appeared responsive to the overall quality of prey in their habitat. Rather than remaining on a prey patch until depletion, predators appeared to vary their patch use with quality of the surrounding environment. When multiple (two) prey patches were available, residence time of predators on a prey patch was shorter than when only a single prey patch was available. Predators seemed to move among the prey patches fairly regularly, dividing their foraging time between the patches and consuming prey from each of them at a similar rate. That predators more than doubled their consumption of prey when we doubled the number of prey (by adding the second patch) is consistent with optimizing behaviors ‐ rather than with an opportunistic increase in prey consumption brought about simply by the addition of more prey. Predators at high density, however, appeared to interfere with each other's foraging success, reflected by their lower rates of prey consumption. Blue crabs appear to forage more successfully (and their prey to experience higher mortality) in prey patches located within 15–20 meters of neighboring patch, than in isolated patches. Our results are likely to apply, at least qualitatively, to other crustacean‐bivalve interactions, including those of commercial interest; their quantitative applicability will depend on the mobility of other predators and the scale of patchiness they perceive.     

Structure, mechanism and mechanical properties of pupal attachment in Greta oto (Lepidoptera: Nymphalidae: Ithomiinae)

The structure and mechanism of pupal attachment are described for the nymphalid Greta oto using electron microscopy, and high‐speed and time‐lapse photography. The cremaster is composed of a 3‐D array of hooked setae that engage with silk fibers spun into layers in a pad on the lower leaf surface. Each seta comprises a shaft terminating in a strongly curved hook, tipped with two lateral barbs. These hook into the silk pad, which is densely laid and built‐up in the central portion, flattening out peripherally. Time‐lapse photography showed that silk pad construction by fifth instar larvae is completed in four distinct spinning movements, producing a random fiber arrangement. It is proposed that such a fiber arrangement provides isotropic strength, giving greater flexibility to the attachment. The cremaster is attached to the silk pad by a series of lateral movements of the pupa's posterior abdomen. This movement, together with the shape of the setal hooks, is thought to be integral to the attachment process. Tensile loading tests showed that attachment failure is due to the breakage of the silk pad, which undergoes gradual destruction before releasing the cremaster. The attachment was found to have high tensile strength and fracture toughness, both of which suggest that it has evolved for the dual purpose of preventing the pupa being pulled from the leaf by a predator and preventing the attachment being weakened by wind, which causes the pupa to swing.     

Structure, composition, and mechanical properties of shark teeth

The teeth of two different shark species (Isurus oxyrinchus and Galeocerdo cuvier) and a geological fluoroapatite single crystal were structurally and chemically characterized. In contrast to dentin, enameloid showed sharp diffraction peaks which indicated a high crystallinity of the enameloid. The lattice parameters of enameloid were close to those of the geological fluoroapatite single crystal. The inorganic part of shark teeth consisted of fluoroapatite with a fluoride content in the enameloid of 3.1 wt.%, i.e., close to the fluoride content of the geological fluoroapatite single crystal (3.64 wt.%). Scanning electron micrographs showed that the crystals in enameloid were highly ordered with a special topological orientation (perpendicular towards the outside surface and parallel towards the center). By thermogravimetry, water, organic matrix, and biomineral in dentin and enameloid of both shark species were determined. Dentin had a higher content of water, organic matrix, and carbonate than enameloid but contained less fluoride. Nanoindentation and Vicker's microhardness tests showed that the enameloid of the shark teeth was approximately six times harder than the dentin. The hardness of shark teeth and human teeth was comparable, both for dentin and enamel/enameloid. In contrast, the geological fluoroapatite single crystal was much harder than both kinds of teeth due to the absence of an organic matrix. In summary, the different biological functions of the shark teeth ("tearing" for Isurus and "cutting" for Galeocerdo) are controlled by the different geometry and not by the chemical or crystallographic composition.     

Seasonal variation in the energy metabolism in an estuarine crab,Chasmagnathus granulata (Dana, 1851)

• 1.1. The effects of seasonal variation on the carbohydrate and lipid metabolism of the Chasmagnathus granulata were investigated.
• 2.2. Glycemia is high in winter and summer and low in spring and fall.
• 3.3. The glycogen content in the hepatopancreas and muscle is higher in fall and winter, and decreases during spring and summer.
• 4.4. The muscle lipids are higher in summer, and decrease during fall and winter whereas hepatopancreas lipids are higher except in the fall.
• 5.5. The crabs show change in the metabolic pattern of lipids and carbohydrates during the seasons of the year.

     

Ontogeny of osmoregulation and salinity tolerance in a mangrove crab, Sesarma curacaoense (Decapoda: Grapsidae)

The grapsid crab Sesarma curacaoense is believed to represent the closest saltwater relative to the ancestor which gave rise to an adaptive radiation of endemic freshwater and terrestrial species on the island of Jamaica. Living in mangrove swamps with variable salinity conditions and showing semiterrestrial behaviour, S. curacaoense exhibits ecological adaptations to non-marine conditions. In laboratory experiments, we studied the salinity tolerance during development from hatching to the end of the first juvenile stage. Successful development through metamorphosis occurred in the full salinity range tested (15-32 per thousand), although mortality was significantly enhanced and development delayed at 15 per thousand. In another series of experiments, we studied the ontogeny of the capability for osmoregulation, which is considered as the physiological basis of osmotic stress tolerance. Our results show that S. curacaoense is from hatching a fairly strong hyperosmoregulator in dilute media. This capability increased gradually from hatching throughout the larval and juvenile development. In seawater (32 per thousand) and at an enhanced salt concentration (44 per thousand), the zoeal stages remained hyperosmoconformers. The capability for hypoosmoregulation in concentrated media appeared first in the megalopa stage and increased thereafter. Adult crabs were observed to be strong hyper-hypo-osmoregulators in a salinity range from at least 1 per thousand to 44 per thousand. The unusually early appearance of strong regulatory capabilities, particularly in dilute media, is interpreted as a physiological preadaptation that should have facilitated the evolutionary process of adaptive radiation in non-marine environments on Jamaica.     

Effect of microcystin on ion regulation and antioxidant system in gills of the estuarine crab Chasmagnathus granulatus (Decapoda,Grapsidae)

The objective of this work was to evaluate mechanisms of microcystin toxicity on crustacean species. Adult male crabs of Chasmagnathus granulatus (13.97+/-0.35 g) acclimated to low salinity (2 per thousand ) were injected with saline (control) or Microcystis aeruginosa aqueous extract (39.2 microg/l) at 24 h intervals for 48 h. After the exposure period, the anterior and posterior gills were dissected, measuring Na(+),K(+)-ATPase and glutathione-S-transferase (GST) activity. Total oxyradical scavenging capacity (TOSC) and lipid peroxides (LPO) content were also determined. Na(+),K(+)-ATPase activity in anterior gills was significantly lower in crabs injected with toxin than in control crabs, while no significant difference in the enzyme activity was detected in posterior gills. Both sodium and chloride concentration in the hemolymph were not affected by toxin exposure. Significant changes in GST activity were detected in posterior gills, with higher values being observed in the toxin-injected crabs. Crabs exposed to microcystin also showed a significant increase in the TOSC value against peroxyl radicals, for both anterior and posterior gills. Lipid peroxides level did not change in both gill types after exposure to the toxin. The increased levels of TOSC suggest the occurrence of a crab response against oxidative stress induced by toxin injection, which prevents lipid peroxidation.     

Temporal genetic homogeneity among shore crab (Carcinus maenas) larval events supplied to an estuarine system on the Portuguese northwest coast

Despite the importance of larval biology in the life histories of many marine animals, relatively little information exists on the dynamics and genetic composition of larval cohorts. The supply of megalopae larvae of the shore crab, Carcinus maenas, was measured on a daily basis during 8 months spread along two larval periods (2006 and 2007) at the Ria de Aveiro estuary, on the Portuguese northwest coast. A total of 10 microsatellite DNA loci were employed to explore the genetic structure, variability and relatedness of temporally distinct megalopal events, selected from the major pulses of supply. Larval variation was also compared genetically with that of a previously studied adult crabs sample, at the same loci, collected in 2006 and 2007 along the Iberian Peninsula. Results revealed a lack of genetic differentiation and identical diversity levels among larval events over time. No evidence of reduced genetic diversity between megalopae relative to the diversity assessed from the pooled sample of adults was found. Moreover, there was no evidence of any family relatedness among larvae from temporal events. The results obtained for C. maenas contradict predictions made by the sweepstakes reproduction hypothesis, in which large variance in reproductive success is expected, which is presumably detectable as sharp genetic discontinuities among separate larval events. Data here indicate conversely a high degree of temporal genetic stability among larval supply to a given estuary under variable oceanographic conditions, consistent with the hypothesis that sampled larvae were drawn from a large number of adults that do not differ in reproductive success.     

Ovigerous-hair stripping substance (OHSS) in an estuarine crab: purification, preliminary characterization, and appearance of the activity in the developing embryos

Ovigerous-hair stripping substance (OHSS) is an active factor in crab hatch water (i.e., filtered medium into which zoea larvae have been released). This factor participates in stripping off the egg attachment structures (i.e., egg case, funiculus, and the coat investing ovigerous hairs) that remain attached to the female's ovigerous hairs after larval release. Thus this activity prepares the hairs for the next clutch of embryos. OHSS activity of an estuarine crab, Sesarma haematocheir, eluted as a single peak on molecular-sieve chromatography, but this peak still showed two protein bands at 32 kDa and 30 kDa on SDS-PAGE. The two protein bands stained with a polyclonal antiserum raised to the active fractions from molecular-sieve chromatography. Moreover, antibodies purified from this polyclonal OHSS antiserum also recognized both the 32-kDa and 30-kDa bands. OHSS immunoreactivity and biological activity were associated with the attachment structures that remained connected to the ovigerous hairs after hatching. In developing embryos, both protein bands could be stained immunochemically at least 10 days before hatching. But OHSS biological activity appeared only 3 days before hatching. The immunoreactive protein bands were not observed in the zoea, but OHSS bioreactivity was present, though greatly reduced. The 32-kDa protein, at least, is probably an active OHSS, and the 30-kDa protein band may also be OHSS-related. The OHSS appears to be produced and stored by the developing embryo. Upon hatching, most of the material may be trapped by the remnant structures, and the remainder is released into the ambient water.     

Biochemical composition of ovary, embryo, and hepatopancreas in the grapsoid crabs Armases cinereum and Sesarma nr. reticulatum (Crustacea, Decapoda)

Biochemical composition of ovary, embryo, and hepatopancreas tissues in wild populations of Armases cinereum and Sesarma nr. reticulatum were monitored during the reproductive season. Total lipid, carbon, nitrogen, C:N ratio, and water concentration of the ovary, hepatopancreas and embryos were quantified over the course of ovarian maturation. Ovary nitrogen concentration decreased as ovaries matured. Ovary lipid and carbon concentration differed significantly over the course of ovarian maturation for both species, but there was no relationship between the concentration or total content of hepatopancreas lipid and the stage of ovarian development in females. Neither species showed a relationship between measures of hepatopancreas lipid and the gonadosomatic index. There was also no simultaneously measurable net decrease in mass of the females' hepatopancreas. Lipid demands of ovarian maturation thus appear to be met in large part by increased dietary intake rather than by substantial draw down of pre-existing lipid stores from the hepatopancreas. While these temperate grapsoid crabs live with putatively fluctuating quality and quantity of food resources, no evidence could be found to demonstrate depletion of lipid concentrations in the hepatopancreas concomitant with ovarian maturation.     

Structure and mechanical properties of hydroxypropylated starch films

Films of acid-hydrolyzed hydroxypropylated pea starch with average molecular weight M w ranging from 3.3 x 10 (4) g/mol to 1.6 x 10 (6) g/mol were prepared from 25% (w/w) solution by casting. The structure of the films was investigated by means X-ray diffraction and calorimetry, evidencing a B-type crystalline structure. In similar drying conditions, 25 degrees C and 40% of relative humidity, the crystallinity varied from 24% for the low molecular weight (A5) to almost none for the highest molecular weight (A160). The influence of the drying temperature was also investigated. A reduction of the crystallinity from 16% to almost none was found when increasing temperature from 25 to 65 degrees C. The glass transition temperature ( T g) at different water contents was determined. The difference of T g between the first and the second scan was interpreted by changes in the water distribution between phases into the B-type crystalline structure. Mechanical properties of the films determined by tensile tests and by DMTA in the glassy state showed no effect of the average molecular weight or of crystallinity. In contrast, thermomechanical experiments by DMTA showed that the average molecular weight of the sample influenced the mechanical relaxation and the moduli in the rubbery state.     

Molt and growth of an estuarine crab, Chasmagnathus granulatus (Brachyura: Varunidae), in Mar Chiquita coastal lagoon, Argentina

Juvenile and adult growth of Chasmagnathus granulatus was studied in the laboratory in terms of molt increment in size (MI) and the intermolt period (IP), comparing data obtained from short‐term (STE) and long‐term (LTE) laboratory experiments. Crabs in a pre‐molt condition were collected for STE, including the entire size range of the species. Larger crabs remained in the laboratory no more than 14 days; the average time to molt was 5.8 ± 3.1 days. We registered the molt of 94 females, 64 males and 34 undifferentiated juveniles and calculated their MI. Moreover, 24 males and four females were reared in the laboratory over 3 years (LTE). Hiatt diagrams did not show sex‐specific differences between juveniles of both sexes, but revealed differences between juveniles and adults in each sex as well as between adults of both sexes. The MI decreased gradually with size; this pattern was described with a quadratic model. The IP increased exponentially with size. The presence of regenerating limbs diminished the MI. The abdomen of females reached its final shape and maximum relative width at functional maturity. Growth curves for both sexes were calculated using the von Bertalanffy model, but this model yielded an underestimation of the actual maximum size of this crab.     

Respiration and osmoregulation of the estuarine crab, Rhithropanopeus harrisii (Gould): effects of the herbicide, alachlor

1. The effects of a sudden decrease in salinity and exposure to sublethal concentrations of the herbicide, alachlor, on osmoregulation and respiration of the crab, Rithropanopeus harrisii, were studied. 2. Crabs were hyperosmotic regulators at salinities below 24 ppt and became hypoosmotic at higher salinities. Upon a salinity decrease from 20 to 1 ppt, crabs adjusted their haemolymph osmolality to a stable hyperosmotic level in 8 hr. Alachlor concentrations to 50 ppm did not affect this adjustment. 3. A salinity decrease from 10 to 0 ppt elevated VO2 and the critical oxygen tension. This response was unaffected by alachlor concentrations as high as 25 ppm.