Effects of ocean acidification on the early developmental stages of the horned turban,Turbo cornutus

To estimate the impact of CO₂-driven ocean acidification on the early life stages of gastropods, the effects of increased partial pressure of seawater carbon dioxide (pCO₂) (800–2,000 μatm) on the early developmental stages and larval shell length of the commercially important gastropod, the horned turban snail, Turbo cornutus were investigated. Increase in experimental seawater pCO₂ had an increasingly negative impact on the early developmental rate; the proportion of embryos or larvae displaying retarded development increased at higher pCO₂. The proportion of embryos that developed to the 4-cell stage at 2 h after fertilization decreased linearly with increasing pCO₂. At ~1,000 μatm pCO₂, retarded development was observed in ~50 % of larvae. No embryos developed to the 4-cell stage at 2,000 μatm pCO₂ within 2 h of fertilization. A similar trend continued until 24–26 h after fertilization; the proportion of larvae attaining veliger stage by 24–26 h also decreased with increasing pCO₂. The shell length of T. cornutus veligers decreased gradually as seawater pCO₂ increased, but markedly decreased in seawater under nearly unsaturated and unsaturated conditions (≤1.04) of the aragonite saturation state (Ω _aragonite). The results indicate that increased pCO₂ seawater has a progressive and acute effect on embryonic and larval T. cornutus, and imply that the extended early developmental period and/or the downsized larval shell produced by ocean acidification will have a negative impact on survival, settlement and recruitment well into the future.     

Acute toxicity of mercury and selenium to Crassostrea gigas embryos and Cancer magister larvae

The possible modification of mercury toxicity by selenium in embryos of the Pacific oyster Crassostrea gigas and the larvae of the crab Cancer magister was investigated. Mercury concentration eliciting abnormal development in 50% of the oyster embryos (EC₅₀) was 5.7 g l^-1 (48 h) and mortality in 50% of the crab larvae (LC₅₀) occurred with 6.6 g l^-1 (96 h). The 48 h EC₅₀ for selenium was greater than 10,000 g l^-1 for oyster embryos and the 96 h LC₅₀ for crab zoeae was 1040 g l^-1. The response from each species, when exposed to both toxicants, revealed, that a high level of selenium (5,000 g l^-1) increased mercury toxicity. Moderate selenium concentrations (10 to 1,000 g l^-1) tended to decrease mercury toxicity, although no statistical verification could be made. The order of administration of toxicants had no effect on the response of Crassostrea gigas embryos. Early developmental stages (8 h) of C. gigas embryos were most sensitive to dissolved Hg; toxicant administration 24 h after fertilization resulted in no apparent abnormalities in development.     

Dissolved saxitoxin causes transient inhibition of sensorimotor function in larval Pacific herring (<Emphasis Type="Italic">Clupea harengus pallasi</Emphasis>)

Herring (Clupea harengus pallasi) spawning sites in Puget Sound, Washington overlap spatially and temporally with blooms of Alexandrium catenella, a toxic dinoflagellate species responsible for paralytic shellfish poisoning. Consequently, newly hatched herring larvae may be regularly exposed to the suite of dissolved paralytic shellfish toxins that are released into the water column from toxic cells during blooms. To date, virtually nothing is known about the impacts of these neurotoxins on early developmental stages of marine fish. In the present study, herring larvae at three ages, 0 days post hatch (dph), 4 dph, and 11 dph, were exposed to dissolved saxitoxin (STX) in 24-h and multi-day exposures. All larvae were examined for sensorimotor function (i.e. spontaneous swimming behavior and touch response). Significant reductions in spontaneous and touch-activated swimming behavior occurred within 1 h of exposure. EC₅₀s at 1 h of exposure were 1,500, 840, and 700 μg STX equiv. l⁻¹ for larvae introduced to STX at 0, 4, and 11 dph, respectively. This progressive age-specific increase in STX-induced paralysis suggests that older larvae were more sensitive to the toxin than younger larvae. Interestingly, herring larvae at all ages exhibited a significant degree of neurobehavioral recovery within 4–24 h of continuous exposure relative to the 1-h time point. This recovery of normal motor behaviors was not observed in previous studies with freshwater zebrafish (Danio rerio) larvae under the same continuous exposure conditions, suggesting that an adaptive detoxification or toxin sequestration mechanism may have evolved in some species of marine fish larvae. Our data reveal that (1) dissolved STX is bioavailable to marine finfish larvae, (2) the toxin is a paralytic agent with potencies that differ between developmental stages, and (3) STX-induced sensorimotor inhibition occurs rapidly but is transient in marine larvae. Collectively, these results suggest that dissolved algal toxins may have important sublethal effects on marine fish populations.     

Embryonic,larval, and post-metamorphic development of the sea urchin <Emphasis Type="Italic">Cassidulus mitis</Emphasis> (Echinoidea; Cassiduloida): an endemic brooding species from Rio de Janeiro,Brazil

Cassiduloids are currently rare irregular echinoids with a highly conserved adult morphology. Aristotle’s lantern is present only during the post-metamorphic stage, and little is known about the early development of species in this group. Cassidulus mitis produces eggs of about 375 μm in diameter, lecithotrophic larvae with four reduced arms with skeletal fenestrated rods, cilia along the body surface, and a ciliated band on arms and lobes. Offspring is brooded among the female spines from embryo to settler’s stage. The echinopluteus larval stage is reached 6 days after fertilization, and the settler’s stage is formed at the age of 17 days. Aristotle’s lantern appears around the thirteenth day of development. The lantern is well developed and functional in settlers. It remains until at least 62 days after fertilization and can be used to acquire food from the environment. The early development of C. mitis is unusual concerning features of typical lecithotrophic larvae (such as reduced arms), but retains some features of planktotrophic larvae (such as skeletal rods and a ciliated band). Regarding egg size, early development in C. mitis seems to be transitioning from facultative lecithotrophic to typical obligate lecithotrophic pattern in echinoid larval evolution.     

Effects of temperature and salinity acclimation of adults on larval survival,physiology, and early development of Lytechinus variegatus (Echinodermata: Echinoidea)

Larval survival and developmental rates of Lytechinus variegatus (Lamarck) were determined as a function of temperature and salinity in two experiments by: (1) directly transferring fertilized eggs to 35, 30, 27.5, 25, 20, 15, and 10S seawater at 18 and 23°C, and (2) acclimation of adult sea urchins to the conditions described above for 1 to 4 wk prior to spawning. Developmental rates and percent survival of larvae prior to metamorphosis decreased at salinities below 35 (Q₁₀ values for metamorphosis=0.380 to 0.384). Temperature and salinity significantly (P<0.05) affected metabolic rates of L. variegatus plutei. These results show that L. variegatus larvae are stenohaline when compared to larvae of other echinoderm species. LC₅₀ values (S), developmental rates, and survival to metamorphosis indicate that acclimation of adult sea urchins to lower salinity prior to spawing and fertilization does not enhance development or survival of embryos exposed to low salinity.     

Accumulation and release of fuel-oil-derived diaromatic hydrocarbons by the polychaete Neanthes arenaceodentata

Male and gravid female Neanthes arenaceodentata (Moore) (Polychaeta: Annelida) were experimentally exposed to a sublethal concentration of No. 2 fuel oil seawater extract for 24 h. Within 1 h both sexes had incorporated, in equal proportions, most of the diaromatic hydrocarbons (naphthalene, methylnaphthalenes) eventually accumulated. No net loss of accumulated naphthalenes was observed during the exposure period. When returned to hydrocarbon-free seawater, male worms slowly released naphthalenes down to undetectable (< 0.1 ppm) levels in 400 h (17 days). Gravid females retained essentially all of the originally accumulated naphthalenes for 3 weeks, at which time they released their eggs during fertilization, with subsequent dramatic decreases in naphthalenes content. Zygotes and trochophore larvae from exposed females contained 18 ppm naphthalenes, accounting for a majority of those amounts initially present in parent females. As trochophore larvae developed into 18-segment juveniles, naphthalenes concentrations dropped to near undetectable levels (0.4 ppm) in 21-day old juveniles (= 18-segment worms). When sampled at the next stage of development (32-segment worms) 5 days later (1050 h after exposure of gravid females), offspring were free of naphthalenes. Results suggest the importance of lipid-rich eggs in the accumulation and retention of diaromatic hydrocarbons by marine polychaetes.     

Développement larvaire de deux Tellinacea,Scrobicularia plana (Semelidae) et Donax vittatus (Donacidae)

Scrobicularia plana Da Costa and Donax vittatus L. were reared in the laboratory through settlement. Fertilizable eggs were obtained by perfusing the ovary with 5% 0.1 M ammonium hydroxyde in sea water. S. plana eggs have a thick chorion, inside which the early larval stages develop; they hatch as straight-hinge larvae more than 60 h after fertilization. This brood protection is considered to be an adaptation to osmotic pressure changes and pollution in the environment. D. vittatus eggs have a very thin chorion and are unprotected. Further development is planktotrophic and very similar for the two species. Under laboratory conditions, the pediveliger stage is attained 3 weeks after fertilization and settlement occurs 1 week thereafter. S. plana spat stop growing until a suitable substratum is available. Meanwhile they undergo a byssus drifting important for postlarval dispersion. Post larvae resume growth as soon as a small quantity of fine sand is added to the rearing jar. The exhalant siphon is developed first, when the post larvae reach a length of 600 m; the inhalant siphon is formed later, at a length of approximately 900 m. Evolution from the larval hinge to the juvenile hinge stage occurs sooner in S. plana than in D. vittatus. Comparison of laboratory larval development with field development indicates that spawning occurs in June and August for S. plana in North Wales (UK).     

Spawning behaviour,egg and larval development of the myctophid fish Benthosema pterotum

The distribution of eggs and early larval stages of the myctophid fish Benthosema pterotum was studied during three cruises to the Gulf of Oman during January to February 1981 and February 1983. The eggs and yolk sac larvae were identified and described after artificial fertilization on board the Research Vessel Dr. Fridtjof Nansen. The mature fish spawn in the Gulf at 300 to 100 m depths during early night. The eggs and smallest larvae were found at 300 to 200 m depths. The eggs hatch within 12 h at 21°C. Hatching occurs before the slightly bouyant eggs reach the upper 50 m of the water column.     

Development of the gooseneck barnacle Pollicipes polymerus (Cirripedia: Lepadomorpha): Fertilization through settlement

Pollicipes polymerus (Sowerby, 1833) is a common barnacle in rocky intertidal zones of the West Coast of North America whose development has been previously undescribed in detail. In the present study, certain aspects of the development of P. polymerus are recorded from fertilization through settlement. In vitro fertilization has been accomplished using already-implanted sperm plus ovulating eggs and oviducal gland fluid. The eggs are small (about 100 μ diameter at fertilization; 90 x 140 μ at first cleavage) and somewhat yolky. Embryos obtained from brooding adults and from in vitro fertilization were reared in glass dishes until hatching. Cleavage was total, asynchronous and unequal. Division of the D macromere lagged behind division of the other blastomeres, and gastrulation occurred by epiboly so that blastoderm surrounded the macromeres. The nauplii hatched an average of 25.4 days (range was 20.3 to 30.1 days) after fertilization, and molted within 1 day to Stage II nauplii. From Stages II through VI the nauplii required food. As they grew from stage to stage, the number of setae per appendage increased. Nauplii have feathery and hispid-type setae, previously described only in Chthamalus aestuarii. In contrast to other pedunculate larvae which have been described, P. polymerus nauplii are small and have specialized feathery setae. Cypris-larva settlement was stimulated only when healthy adult peduncles were available, thus showing an affinity for its own species.     

Digestive enzymes in the ontogenetic stages of the southern king crab, Lithodes santolla

The early ontogenetic stages of the sub-Antarctic king crab Lithodes santolla were analyzed for the presence and activities of a set of important digestive enzymes. The eggs and non-feeding larvae (zoea I-III, megalopa) showed high activities of esterases, phosphatases, and exopeptidases indicating the enzymatic ability to utilize endogeneous yolk reserves. SDS-PAGE showed a continuous decrease of proteins or proteids in the range of 59–81 kDa during ontogenetic development from the eggs through the zoeal stages to the first juvenile crab stage, CI. This reduction reflects the degradation of storage compounds during lecithotrophic larval development. Activities of the endopeptidases, trypsin and chymotrypsin, were low in eggs and larvae but increased significantly in the first juvenile crab stage. These enzymes typically facilitate the first steps of proteolysis in the extra-cellular spaces of the midgut gland and in the stomach. Their scarcity indicates that the larvae of L. santolla are physiologically not prepared to digest external food. This ability seems to appear first in the CI stage. Extracts of juvenile midgut glands and the gastric fluids of adults showed high activities of a variety of digestive enzymes including phosphatases, carbohydrases, as well as endo- and exopeptidases. High activities of digestive enzymes in adults may compensate for scarce food supply and rate-limiting low temperatures in the predominantly sub-Antarctic habitats of L. santolla.     

Physiological and biochemical aspects of embryonic and larval development of the winter flounder Pseudopleuronectes americanus

Eggs and larvae of the winter flounder Pseudopleuronectes americanus Walbaum were hatched and raised in the laboratory under controlled conditions. Biochemical composition was measured during development and found to be similar to that of other species: 65 to 80 percent protein, 15 to 30 percent fat, and 0 to 5 percent carbohydrate. Ash content was 7 to 10 percent of dry weight. The chorion comprised more than half of the weight of an egg and the data suggested that it was possibly a source of nutrition to the developing embryo. The sequence of utilization appeared to be carbohydrate and then protein to hatching, lipid, mixed lipid and protein, the predominantly protein until feeding began. Carbohydrate was accumulated at first feeding and depleted when growth began. Protein and lipid were deposited in approximately constant proportions. Respiration rates of eggs were low, 0.002–0.015 l O₂ egg^-1 h^-1, but rose gradually from fertilization to hatching. Respiration rates of early larvae were from two to eight times that of eggs (0.033–0.131 l O₂ larva^-1 h^-1). Variation in larval respiration rates indicated a three-fold difference in rate according to level of activity. Eggs excreted ammonia at an increasing rate from fertilization to hatching. Larvae excreted ammonia, primary amines, and other unidentified organic nitrogenous substances. Rates of excretion and proportions of excretory products varied with stage of development. Primary amine excretion was variable and a major component in early stages. Ammonia-N excreted was two to 20 times primary amine N excreted. Unidentified substances were the predominant form of N excretion during early feeding. Ammonia accounted for most of the N excreted in older larvae. Early specific growth rates were 2.1 and 5.5%. Net caloric conversion and net and gross nitrogen efficiencies were low in first feeding larvae compared to adult fishes (32.2, 27.7, and 10.7% respectively).Contribution no. 5071 from the Woods Hole Oceanographic Institution     

Carbofuran induced changes in breeding of a freshwater fish,Labeo rohita (Hamilton)

The effect of carbofuran, a carbamate pesticide, on the breeding performance of the freshwater fish Labeo rohita (Hamilton) has been investigated in the present study. Breeding of L. rohita was conducted after treatment with three sublethal concentrations, i.e. 0.06, 0.15 and 0.30 mg L^?1 of carbofuran for 96 h. The present investigation showed that the number of total eggs and total amount of eggs (litre per kg body weight) decreased significantly (p < 0.01) at all concentrations of carbofuran in comparison to controls, while the reduction in fertilization percentage at all concentrations of carbofuran was not significantly different from controls. The reduction in hatching percentage, the expected number of hatchlings, and the expected number of hatched larvae were significantly (p < 0.01) different between treated and control groups at all carbofuran concentrations. No significant differences for the 96 h survivability of hatched larvae were reported at all concentrations of carbofuran.     

Gametogenesis,embryogenesis, and larval features of the oviparous sponge <Emphasis Type="Italic">Petrosia ficiformis</Emphasis> (Haplosclerida,Demospongiae)

This study describes the reproductive cycle of Petrosia ficiformis and documents for the first time embryogenesis in an oviparous haplosclerid demosponge. Gonochoric adults, occurring in 2:1 female/male ratios, spawned in late autumn, after a 7-month long oogenesis and a 2.5-week-long spermatogenesis. Following a remarkable migration, the oocytes were released as 250 μm eggs bearing attached polar bodies and a thin mucous cover. Round-headed spermatozoa with three large mitochondria, and many proacrosomal vesicles fertilized the eggs externally. A fertilization membrane appeared around the zygotes. Nearly equal and total cleavage led to a stereoblastula that subsequently became an entirely ciliated larva. The larva alternated spherical and multilobate body shape and consisted of undifferentiated cells. It had poor abilities to swim and glide, probably experiencing passive dispersal and unselective attachment. Settlers developed choanocyte chambers after 1.5 months. Symbiotic microbes were absent from gametes and larvae, being necessarily acquired from the ambient at each sponge generation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effects of copper on the coral-reef echinoid Echinometra mathaei

The effects of copper-enriched seawater on fertilization success, early cleavage, larval skeletal development, and survivorship of adults of the coral-reef echinoid Echinometra mathaei (de Blaiville) were tested. Fifty percent reduction of fertilization success and cleavage success to the 8-cell stage occurred in 0.18 and 0.42 mg added Cu⁺⁺/l, respectively. Adults had respective 48- and 96-h TL₅₀ values of 0.54 and 0.30 mg/l. Larval skeletal development was suppressed in 0.02 mg/l. The latter process appears to be very sensitive and may be the most suitable for assessing effects of stress on this species.Contribution No. 68, University of Guam Marine Laboratory.     

脱氧雪腐镰刀菌烯醇、黄曲霉毒素B_1和玉米赤霉烯酮对秀丽隐杆线虫的联合毒性研究

脱氧雪腐镰刀菌烯醇(DON)、黄曲霉毒素B_1(AFB_1)和玉米赤霉烯酮(ZEN)是最为常见的粮食真菌毒素,易共存于谷物产品和动物饲料中,而目前对其联合毒性的研究较少,且研究结果并不完全一致。为探究DON、AFB_1和ZEN的联合毒性作用,本论文以秀丽隐杆线虫(C.elegans)为模型,分别评估了毒素混合物AFB_1+DON、AFB_1+ZEN、DON+ZEN和AFB_1+DON+ZEN对C.elegans的生长发育(体长)和生殖能力(产卵量)的毒性作用,并用Chou-Talalay模型来判定毒素混合物的相互作用类型。研究表明,AFB_1、DON和ZEN单独染毒C.elegans时,其毒作用强弱为AFB_1ZENDON。联合染毒时,AFB_1+DON对C.elegans产生协同作用,而DON+ZEN则产生拮抗作用;AFB_1+ZEN对体长(24 h)和产卵量的毒作用随着暴露浓度的增加,由弱拮抗变为协同作用,而在毒素暴露48 h后,对线虫的生长发育呈协同作用;AFB_1+DON+ZEN除在EC50-24 h和EC75-24 h时对体长产生明显的毒性增强作用外,其他普遍表现出拮抗作用。由此表明,DON、AFB_1和ZEN对C.elegans的联合毒性作用与剂量和时间相关。     

Reproduction and development of the polychaete Stauronereis rudolphi,including a summary of development in the superfamily Eunicea

The life history of the dorvilleid polychaete Stauronereis rudolphi (Delle Chiaje) was studied through the F₂ generation under laboratory conditions. The species released its gametes into the water where fertilization occurred. The pelagic larvae swam for one week before settling. Segments were added successively and sexual maturity was reached in one year of laboratory culture. Spawned adults recovered and began feeding a few days after spawning. The culture techniques and foods used to rear developing eggs, larvae, juveniles, and adults were described. Comparisons were made with other developmental studies in the superfamily Eunicea.This study was a portion of a thesis submitted in partial fulfillment of the requirements for the degree of Master of Arts in the Department of Biology, California State College at Long Beach. It was in part supported by research grant WP-00531-02 from the National Institute of Health, U. S. Public Health Service, under supervision of Dr. Donald J. Reish. Contribution No. 1 of the Ira C. Darling Center for Research, Teaching and Service, University of Maine, Maine 04573, U.S.A.     

Biological effects of oil on early development of the Baltic herring Clupea harengus membras

The effects of petroleum hydrocarbons from two crude oils and one fuel oil (No. 1) were studied on the ontogenic development of the Baltic herring Clupea harengus membras L. Herring eggs exposed to water-soluble fractions of the oils at the time of fertilization showed no decrease in numbers of fertilized eggs compared to eggs exposed 6 or 72 h after fertilization. During embryongenesis, treatment with 3.1 to 8.9 ppm or 3.3 to 11.9 ppm total oil hydrocarbons from light fuel oil and the two crude oils respectively, gave rise to alterations in embryonic activity, decreased heart rate, and premature or delayed hatching. Although many larvae hatched from eggs exposed to contaminated water (3.1 to 11.9 ppm total oil hydrocarbons), the majority of the (70 to 100%) were malformed or dead 1 day after hatching. Exposure of eggs to 5.4–5.8 ppm total oil hydrocarbons resulted in significantly (P<0.001) decreased lengths of the larvae. Increased temperature (from 9° to 14°C) aggravated the effects of the oils. The results are discussed in relation to the potential effects of oil spills and chronic oil pollution on fish eggs and larvae in the Baltic Sea.     

Growth, carbon, nitrogen and caloric content of Solea senegalensis (Pisces: Soleidae) from egg fertilization to metamorphosis

Eggs and larvae of the Senegal sole, Solea senegalensis Kaup, were reared from fertilization until the end of metamorphosis, which occurs by Day 17 after hatching at 19.5 °C. Changes in energy content and biomass quality were studied in terms of dry weight and of carbon, nitrogen and energy content. S. senegalensis spawned eggs of about 1 mm diameter which hatched 38 h after fertilization. Average dry weight of individual eggs was 46 μg, the chorion accounting for about 18% of total dry weight. Gross energy of recently fertilized sole eggs was approximately 1 J egg⁻¹. From fertilization to hatching, eggs lost 8% of their total energy (chorion not included). After hatching, larvae lost 14% of their initial energy until the start of feeding which occurred about 48 h afterwards. The principal components catabolized during embryogenesis were carbon-rich compounds that decreased by 26%, while nitrogen-rich compounds decreased by only 10% and were practically unaltered from hatching to the start of feeding. Feeding larvae displayed constant growth during the period studied (specific growth rate on a dry weight basis was 0.26 d⁻¹). The relative proportion of carbon and nitrogen content revealed an accumulation of high energy compounds in the days before metamorphosis. By Day 14, the energy content reached values similar to those of recently hatched embryos, but decreased again during metamorphosis. Received: 10 June 1998 / Accepted: 28 January 1999     

Sequential utilization of free amino acids, yolk proteins and lipids in developing eggs and yolk-sac larvae of barfin flounder Verasper moseri

Changes in the contents of free amino acids (FAA), M_r 170,000 lipovitellin (oLv B), that is the major yolk protein in ovulated eggs, and lipids were measured in developing eggs and yolk-sac larvae of barfin flounder (Verasper moseri) to elucidate the sequential utilization of these nutrient stocks before the first feed. Hatching takes place on the 10th day after fertilization at a water temperature of 8°C, and the hatched larvae absorb almost all of their yolk masses within 21 days after fertilization. The total FAA content showed no change during the first 4 days, then decreased to about 13% of the initial level by the 13th day after fertilization. During the consumption of FAA, non-essential amino acids tend to decrease earlier than essential amino acids. The native molecular weight and immunoreactivity of M_r 170,000 oLv B did not show any change during the first 16 days after fertilization, though the sodium dodecyl sulphate-polyacrylamide gel electrophoresis patterns of oLv B showed a serial change. The oLv B contents, measured by quantitative immunodiffusion using antiserum against oLv B of ovulated eggs, were approximately stable during the 13 days after fertilization, then decreased rapidly until the end of yolk-sac absorption. Phospholipids (PL), which seem mostly to bind with lipovitellin-proteins, decreased gradually after hatching coincident with the decrease in oLv B and M_r 330,000 lipoprotein. From these results, we conclude that there are three periods for sequential nutrient utilization in barfin flounder embryos and larvae: (1) pre-FAA utilization period, 0-4th day; (2) FAA utilization period, 4th-13th day; (3) oLv B and PL utilization period, 16th-21st day post-fertilization.