胡子鲶的胚胎和幼鱼发育的研究

胡子鲶(Clarias fuscus)的胚胎和幼鱼的发育过程与蟾胡子鲶(C.batrachus)类似,器官分化时间和大小比例则有一定的差异。胡子鲶卵呈球形,富含卵黄,卵径1.7—1.9毫米;出膜仔鱼全长4.8—5.1毫米,比蟾胡子鲶约大1/3;幼鱼卵黄囊被吸收消失的时间比蟾胡子鲶迟1—2天。心脏的出现和搏动与开始出现血液循环的间隔时间只有3—5小时;耳石的出现几乎和心脏的出现处在同一时间内,与白鲢有较大的差异。在仔鱼前期居维氏管明显,但出现的时间比蟾胡子鲶稍迟些。仔鱼出膜形式与蟾胡子鲶不同,也与一般鱼类出膜形式不同。仔鱼是以腹部卵黄囊顶破卵膜,以卵黄囊先出膜,一般鱼类多数是以头部或尾部先出膜的。在水温28.5—31℃的条件下,从受精到孵化出膜的时间为28小时25分;幼鱼期共历时12—15天。       

Embrionary and larval development of the marine clam Tivela mactroides (Bivalvia: Veneridae) in Zulia State, Venezuela

The marine clam, Tivela mactroides, from Ca?o Sagua beach, Venezuela, was spawned and reared under laboratory conditions to monitor its early development. Spawning was spontaneous but in some cases it had to be induced by the additon of eggs and sperm. After fertilization, the embryonic development occurred at 5 hr approximately. Trochophore larvae were observed between eight and ten hours later. Straight-hinged veliger stage appeared 15 hr after fertilization. Transition from veliger stage to the umbo stage occurred about eight days after fertilization. Pediveliger stage was observed 22 days after fertilization. Metamorphosis of T. mactroides was not successful under our laboratory conditions; probably the bacterial contamination and subsequent mortalities were important factors constraining the final phase of the larval cycle. However, in a few cases young individuals were observed. We suspect that this was due to unfavorable conditions (e.g.: bacterial contamination, unsuitable food availability, etc.) and the broad variation in developmental times, suggesting that these stages might be particularly sensitive to environmental changes. These results may not necessarily reflect what happens under natural conditions.     

Assortative fertilization and selection at larval stage in the mussels Mytilus edulis and M. galloprovincialis

Assortative mating (prezygotic isolation) and reduced hybrid fitness (postzygotic isolation) are typically invoked to explain the stability of hybrid zones. In the tension zone model, these factors work in opposition to migration, which promotes genetic homogeneity. Many marine animals migrate over long distances through a planktonic larval stage. Therefore, strong reproductive isolation is needed to maintain stable marine hybrid zones. However, surprisingly little is known about mating preferences and hybrid fitness in marine organisms. Smooth-shelled mussels (Mytilus spp.) form a well-known species complex, with hybridization over extensive areas such as the contact zone of M. edulis and M. galloprovincialis around European Atlantic coasts. This paper reports direct experimental evidence of assortative fertilization, hybrid larval inviability, and early heterosis for growth rate in M. edulis and M. galloprovincialis. Four crosses between pure M. edulis and M. galloprovincialis were analyzed with a new polymerase-chain-reaction-based diagnostic marker. Gamete competition between taxa was allowed in two out of the four crosses. Genotype frequencies observed at an early stage (36 h after fertilization) unambiguously revealed assortative fertilization when gamete competition was allowed. A significant reduction in hybrid viability was subsequently observed during the larval stage. At the same stage an antagonistic effect, heterosis, was observed on growth rate. However, even if heterosis is observed in the F1, it is expected to vanish in subsequent hybrid generations. Although specialization for different habitats and asynchronous spawning have been mentioned as factors contributing to the maintenance of the blue mussel hybrid zone in Europe, we argue that assortative fertilization and reduced hybrid fitness are important factors that also contribute to the stabilization of this zone. These results emphasize that multiple factors may act concomitantly in a barrier to gene flow, especially in complex life cycles. Furthermore, they show that assortative mating through gamete preference, as already demonstrated for sea urchins, may play a role in speciation processes taking place in the sea.     

Larval growth and survival of Echinometra lucunter (Echinoidea: Echinometridae) fed with microalgae Chaetoceros gracilis and Isochrysis galbana

Thirty sexually mature sea urchins (Echinometra lucunter; diameter 45.8 +/- 17.5 mm) were collected at Macanao, Margarita Island, Venezuela (11 degrees 48'29" N / 64 degrees 13'10" W). They were injected potassium chloride (50 M) directly into the celomic cavity. After two minutes 90% spawned (17 females and 10 males), the others never spawned. Fertilization was 87.0 +/- 12.6% (1:100 oocytes/sperm) at 29 +/- 2 degrees C. The fertile eggs were placed in three treatment gropsu with nine containers (18 liters; 2 eggs/ml) each, all with bottom aeration. Treatments were: Chaetoceros gracilis; Isochrysis galbana, and a mixture of both microalgae (respectively: 20 000 and 60,000 cell/ml for each microalgae, 1:1 for the mixture). Salinity, pH, temperature and larval survival were determinated daily. The study ended when the post-metamorphic phase was completed. The embryonic development time was 16.3 +/- 0.2 h until the prism stage at pH 8.4 +/- 0.1; 38 +/- 1 psu and 28 +/- 1.4 degrees C. The two-arms larval stage was reached at 24 h: 33 min, with a total length of 190 +/- 16.3 microm fed on C. gracilis, 152 +/- 19.0 microm with I. galbana and 182.4 +/- 14.1 microm with the mixture. The larvae next to metamorphosis reabsorbed the arms and had the characteristic shape of juvenile urchins at 12 days with 670.2 +/- 22.2 microm fed on C. gracilis, 665 +/- 12.1 microm fed on I. galbana and 670 +/- 14.1 microm fed on the mixture. The accumulated survival to the juvenile stage was 14.7 +/- 3.8% when fed on C. gracilis, higher than the other treatments (5.4 +/- 1.2; 14.0 +/- 2.6). E. lucunter is an excellent prospect to be commercially cultured because of its short embryonic (16 hours) and larval development time (12 days) and good survival rate when fed on monoculture (C. gracilis and I. galbana) or mixed diet (we recommend C. gracilis).     

Sperm surface proteins persist after fertilization

Certain sperm components labeled with fluorescein isothiocyanate or its radioactive derivative, 125I-diiodofluorescein isothiocyanate (125IFC), are transferred at fertilization to the egg, where they persist throughout early cleavage stages at a localized site in the embryo cytoplasm (Gabel, C. A., E. M. Eddy, and B. M. Shapiro, 1979, Cell, 18:207-215; Gundersen, G. G., C. A. Gabel, and B. M. Shapiro, 1982, Dev. Biol., 93:59-72). By using image intensification we have extended these observations in the sea urchin to the pluteus larval stage, in which greater than 60% of the embryos have localized fluorescent sperm components. Because of the unusual persistence of the sperm components in the embryo, a characterization of the nature of the labeled species in sea urchin sperm was undertaken. Approximately 10% of the 125IFC was in sperm polypeptides of Mr greater than 15,000. These proteins were on the sperm surface as shown by their sensitivity to externally added proteases. The remainder of the 125IFC in sperm was in several low- molecular-weight species, none of which was 125IFC-derivatized phospholipid. To determine if any labeled sperm polypeptides remained intact in the embryo after fertilization, 125IFC-labeled sperm proteins were recovered from one-cell and late gastrula stage embryos by using an anti-IFC immunoadsorbent. Most of the labeled sperm proteins were degraded shortly after fertilization; however, distinct sets of labeled polypeptides were recovered from both one-cell and gastrula stage embryos. Six of the labeled polypeptides recovered from both embryonic stages had identical SDS gel mobilities as labeled sperm polypeptides. Other polypeptides in the embryos appeared to arise from limited proteolysis of sperm proteins. Thus, in this physiological cell fusion system, individual sperm proteins are transferred to the egg at fertilization, and some persist intact or after specific, limited degradation long after gamete fusion, until at least the late gastrula stage.     

Juvenile production of the red sea urchin Strongylocentrotus franciscanus (Echinodermata: Echinoidea) in Baja California, Mexico

The red sea urchin Strongylocentrotusfranciscanus (Agassiz 1863) is harvested commercially in Baja California, Mexico, since 1970; however, in the last ten years the capture per unit effort (CPUE) has decreased from 310 kg/fishing unit/day to 120 kg/fishing unit/day. For this reason, actions were taken to develop a culture technology allowing massive production of juveniles for re-stocking natural populations or for growing them commercially. We summarize some of the basic studies and main achievements in this effort. In Baja California, considerably faster larval development (approximately 21 days) has been attained than in the US northwest coast (62 days). Spawning of red sea urchins was routinely induced with KCI while egg fertilization was performed using a 100,000-sperm/ml solution. Six microalgae species were tested and Rhodomonas sp. produced the best larval development. The mean survival rate at the end of the larval period was 25%, but results varied widely with bactch. From the feed ratios tested, best results were obtained using 7000 cel/ml during the first week of larval development, followed by 10,000 cel/ml during the second and 15,000 cel/ml during the third week. KCl proved the most consistent metamorphic inducer, regularly yielding metamorphosis percentages higher than 90%. Metamorphosis was considered complete when the functional jaw that juveniles use for first benthic feeding appeared (as soon as 20 days after induction). With this method several thousands of red sea urchin juveniles were produced. They reached up to 1.5 mm in size during the first 50 days of culture after metamorphosis, showing the great potential for mass production of this species in the laboratory.     

Exogastrulation and interference with the expression of major yolk protein by estrogens administered to sea urchins

Although estrogens have been detected in some echinoderm species, their role is not clearly understood; so we examined the effects of estrogens administered to sea urchin embryos and larvae. A typical malformation was exogastrulation, induced by the exposure to ethynylestradiol (EER) in a defined period of 12 h from 12 h after fertilization (HAF). Morphogenesis for gastrulation was delayed in the treated embryos: protrusion of the archenteron started at 30 HAF when gastrulation had already finished in normal embryos. Exogastrulation induced by EER was cancelled by the antiestrogen chemical, ICI182,780. Feeding larvae were less sensitive to estrogens than those in early embryogenesis and, at certain concentrations, developed without abnormal morphology. The effect of estrogens was examined at the level of gene expression of the major yolk protein (MYP). MYP expression started during the larval stage and was suppressed by estrone at the six-armed stage, but not by β-estradiol, and in later stage larvae, the expression was not affected by treatment with either estrogen. Estrogens affect sea urchins in the early stage of embryogenesis, leading to abnormal morphogenesis and interference with gene expression.     

Laboratory spawning, larval development, and metamorphosis of the limpets Lottia digitalis and Lottia asmi (Patellogastropoda, Lottiidae)

Abstract. This study describes and compares laboratory spawning, larval development, and metamorphosis in the patellogastropod limpets Lottia digitalis and Lottia asmi. Both species were dioecious and freely spawned their gametes, which were fertilized externally. Eggs from L. digitalis and L. asmi averaged 155 and 134 μm in diameter, respectively. Early cleavage patterns were typical of other patellogastropods. Swimming trochophore larvae had developed ∼ 15 hours after fertilization, and ultimately developed into lecithotrophic veliger larvae that reached metamorphic competence at 5.25–5.5 days after fertilization (13°C). Food particles were frequently visible in the gut of newly metamorphosed individuals one day after settlement, and adult shell growth was typically initiated within 2–4 days of settlement. Small egg size in L. asmi , relative to other eastern Pacific lottiids, may be directly related to the need for high fecundity in this small-bodied species; however, developmental information is available for relatively few lottiid species. Because broadcasting lottiids do not secure egg masses in safe microhabitats for development, this reproductive mode may have been conducive to their ecological radiation into novel habitats.     

Effects of retinoic acid and dimethylsulfoxide on the morphogenesis of the sea urchin embryo

Sea urchin embryos of the species Paracentrotus lividus were treated continuously with different concentrations of all-trans retinoic acid (RA) or dimethylsulfoxide (DMSO) at different developmental stages. A delay in embryonic development was observed when embryos were cultured in the presence of 2x10^?5 M RA, between 1 and 12 hours of development. Hence, at 48 hours of development, while control embryos had reached the pluteus stage, RA-treated embryos were at the prism stage. At 72 hours of development RA-treated embryos recovered and continued normal development reaching the pluteus stage. No effect was observed when treatment was performed before 1 hour or after 12 hours of developmet. DMSO treatment had no effect on normal sea urchin embryo development, although we observed that pigment cells, clearly visible at the pluteus stage, become visible earlier with respect to control embryos. This report confirms the advantages that the sea urchin embryo offers for the study of problems in cellular and developmental biology.     

RNA/DNA ratio as an index of physiological condition of Colossoma macropomum and Piaractus branchypomus (Pisces: Characiformes) during embryonic development

We evaluated RNA/DNA ratio as an index of physiological condition during larval development of a hybrid between the fishes Colossoma macropomum (cachama) and Piaractus brachypomus (morocoto). The samples were obtained by induced reproductive technology and the eggs were maintained in acrylic conical incubator with a continuous waterflow. Embryonic development, from egg fertilization to cell division and hatch out, took 12 hours 20 minutes at 29.5 degrees C, dissolved oxygen contents of 6.0 ppm and pH 7.5. Nucleic acids quantification was determined by fluorometry with ethidium bromide and Hoechst 33258 dyes. We observed significant changes of RNA/DNA ratios during all stages of the embryonic larval development. Therefore, RNA/DNA relation is an useful technique to evaluate physiological condition in short period and could be utilized as nutritional condition and/or instantaneous growth for routine check to verify the health status in early life of cultivated species.     

Inhibition by aphidicolin of cell cycle progression and DNA replication in sea urchin embryos.

We have recently found that aphidicolin, a tetracyclic diterpene-tetraol produced by several fungi, blocks DNA synthesis of sea urchin embryos by interfering with the activity of DNA polyermase alpha. These cells fail to proliferate in the presence of aphidicolin. In continuation of these studies, we determined the drug-sensitive stage in the first cell cycle of the sea urchin Clypeaster japonicus embryo. In continuous exposure to aphidicolin (2 micrograms/ml) from five minutes after fertilization, mitotic division of the embryo was completely suppressed. Embryos were exposed to the drug at progressively later intervals and their capability for cytokinesis was examined. Evidence was thereby obtained that aphidicolin acts at the S-period to inhibit DNA synthesis resulting in developmental arrest of the embryo.     

Fertilization in landlocked sea lamprey: storage of gametes, optimal sperm: egg ratio, and methods of assessing fertilization success

Ovulated, unfertilized eggs of sea lamprey Petromyzon marinus could be stored for 1 day at 15° C without significant loss of fertilizing ability. After 2 days storage most eggs could still be fertilized. Lamprey semen could be stored up to 1 day. Thereafter, a decrease in sperm fertilizing ability occurred, accompanied with a decrease in sperm motility. Unlike teleost fish, sea lamprey eggs could still be fertilized after 1 h contact with water. This extended time of gamete fertility after release into water may help to account for the reproductive success of this species. Maximal fertilization rates were obtained at a sperm: egg ratio of 50 000, a ratio recommended for studies on fertility of individual males. Assessing fertilization success 3 min after fertilization (at cytoplasmic bleb stage) or 5 h after fertilization (at two–cell embryo) was strongly correlated ( r =0·92 and 0·98) with estimation and fertilization success at hatching. These results offer improvement in artificial fertilization techniques under laboratory conditions and provide new information on the biology of fertilization in sea lamprey.     

Sea bass Dicentrarchus labrax nervous necrosis virus isolates with distinct pathogenicity to sea bass larvae.

Reproduction of nodavirus disease was performed by experimental infection of sea bass eggs during fertilization or at larval stage 4 with 2 genetically distinguishable nodavirus strains (Sb1 and Sb2) isolated from sea bass collected along the Atlantic and Mediterranean French coast. The pathogenicity of the virus strains was assigned after detection of the virus by ELISA and immunohistochemistry (IHC). The Atlantic (Sb1) strain was more pathogenic than the Mediterranean (Sb2) strain during the fertilization step whilst both strains were pathogenic following experimental exposure of 4 d old larvae. Virus lesions developed in the brain 4 to 6 d following experimental exposure. Experimental ELISA proved very sensitive for detecting the nodavirus in Sb1 or Sb2 experimentally infected larvae, as well as in naturally infected sea bass larvae collected in French hatcheries or in barramundi larvae reared in the Pacific area. The development of an ELISA specific for the 2 nodavirus strains isolated from the sea bass should be useful for the detection of the virus, in addition to other techniques recommended by the Office International des Epizooties (OIE).     

Larval development and post‐larval growth of Branchiomma bairdi (Annelida: Sabellidae) from a Mediterranean population

Branchiomma bairdi is a Caribbean fan worm introduced in several localities worldwide, including the Mediterranean Sea, where the species’ range has rapidly expanded. Reproduction in B. bairdi was previously investigated in both extra‐Mediterranean and Mediterranean areas, but no information is available on larval development and post‐larval growth. In the present article, we examined these features for a population from the Mar Grande of Taranto (Ionian Sea). The species is hermaphrodite, and fertilization occurs in situ. Mucus seems to play an important role in fertilization, and also in preserving eggs before fertilization. The trochophore stage develops within the mucus and after hatching, larvae swim for about 3 d before settlement. The trochophore showed a distinct prototroch and two red dorsolateral larval eyes. The pelagic stage takes only 96 h even though prototroch is maintained after settlement, disappearing at 5 d, when larvae showed three chaetigers and branchial crown consisted of four radioles. Some interesting observations concerning changes in the morphology of chaetae and in the number of uncini during growth are also reported, together with discussion of the development of stylodes, an important diagnostic feature in Branchiomma species identification.     

The ecological and anatomy of a new species of aeolid opisthobranch mollusc; a predator of the scleractinian coral Porites

A new species of aeolid opisthobranch (family Tergipedidae) is described from Tanzania. It lives and feeds on the scleractinian coral Porites somaliensis. Studies on its development show that it can reach sexual maturity three weeks after hatching. The larval stage is extremely abbreviated; the veliger settles ten minutes after hatching, and within ninety minutes of hatching has assumed a slug-like form.     

Larval development of certain gamete-spawning scleractinian corals

Embryogenesis and larval development were documented in 19 species of hermatypic scleractinians which release gametes during the summer coral spawning season on the Great Barrier Reef. Cleavage of fertilized eggs began approximately 2 h after spawning in all species, and gave rise to blastulae after 7–10 h. Endoderm formation in Platygyra sinensis was by invagination, and this appeared to occur in all species studied. All species observed at 36 h after spawing were mobile and full mobility was reached by 48 h. Settlement of planulae placed in aquaria occurred between 4 and 7 days after fertilization. These results suggest that larval corals produced by most gamete-releasing coral species are likely to be dispersed away from the parent reef.     

SPAWNING AND LARVAL DEVELOPMENT OF CLYPEOMORUS CLYPEOMORUS JOUSSEAUME 1888 IN WALTAIR COAST

Clustering and pairing of the two sexes of the prosobranch Clypeomorusclypeomorus Jousseaume, 1888 was observed during the spawningperiod. The eggs are released in the form of a thin filamentwhich folds in a definite fashion and forms a ribbon. Thereare two larval stages in development, the trochophore and theveliger. The maximum number of trochophores wasobserved 13 hoursand the maximum number of veligers 18 hours after fertilization.The swimming-crawling stage was also observed. (Received 5 July 1979;