Water and salt balance of halibut eggs and larvae (Hippoglossus hippoglossus)

Eggs of halibut [Hippoglossus hippoglossus (L.)] have a negative buoyancy in sea water of 35 S, in contrast to eggs of most flatfish species. The cause of this was investigated. The osmolality of the yolk is 350–420 mOsm during embryonic development. This is within the range for marine teleost serum and for yolk of pelagic eggs. Concentrations of major inorganic ions are comparable with those of pelagic plaice eggs [Pleuronectes platessa (L.)]. The values for Na⁺, K⁺ and Cl^- are 6, 85, and 64 mmol · (1 H₂O)^-1 after fertilization, and at the time of hatching the corresponding values are 17, 11, and 80. Large amounts of other inorganic constitutents are excluded for osmotic reasons. Malfunction in the regulation of osmolality or of inorganic constituents is thus unlikely to be the cause of negative buoyancy. The relative dry weight of the chorion (egg shell) in halibut eggs is less than in several pelagic egg types, excluding the chorion as the main contributor of negative buoyancy. It is concluded that a high content of organic matter in the rest of the egg is the cause of the negative buoyancy.     

Effects of tidal fluctuations of salinity on pericardial fluid composition of the American oyster Crassostrea virginica

Crassostrea virginica Gmelin were subjected to simulated tidal fluctuations of salinity, and the subsequent effects on osmotic and ionic composition of the pericardial fluid, body water and valve movements were investigated. Ambient salinity fluctuation patterns of 20-10-20, 15-10-15 and 10-5-10 were simulated during 24.8-h periods. An additional 10-5-10 S experiment was performed using a dilution water approximating the ionic composition of Mississippi River water with regard to Mg⁺⁺, Ca⁺⁺ and SO ₄ ⁼ , instead of deionized water. Finally the effects of a 2-week diurnal fluctuation pattern between 20 and 10 S were investigated with respect to pericardial fluid composition. Pericardial fluid osmolality, concentrations of Cl^-, Na⁺, Mg⁺⁺, K⁺, Ca⁺⁺ and ninhydrin-positive substances (NPS) were analyzed periodically throughout all experiments. Pericardial fluid osmolality was slightly hyperosmotic as ambient water salinity decreased during a cycle, and then became slightly hyposmotic as ambient salinity increased. In the 2-week experiment, pericardial fluid osmolality tracked ambient seawater closely through Day 5, but became more intermediate between high and low seawater values as the experiment progressed. Similar patterns during fluctuations of salinity were observed for Na⁺, Cl^-, Mg⁺⁺ and Ca⁺⁺. Pericardial fluid K⁺ levels did not track ambient seawater as closely as did other ions. The ionic composition of dilution water had little effect on the osmotic or ionic response of the oyster's pericardial fluid. Pericardial fluid NPS level varied inversely with salinity during the 20-10-20 cycle. During the longterm fluctuation experiment, NPS values gradually decreased over the 2-week period compared to constant salinity control values. Percent body water also varied inversely with ambient salinity. Solute movement accounted for most of the change in pericardial fluid osmolality during the simulated cycles with water movement responsible for 1 to 11%. Water movement contributed more to the change of pericardial fluid osmolality during the decreasing salinity phase than the increasing phase of a given cycle. During 20-10-20 S cycles, oyster valves remained open 56% of the time (n=23). In contrast, when salinity was abruptly changed from 20 to 10 within 5 min, valve closure occurred in 4.8±0.3 min (n=20). Valves did not reopen for 19.3±1.2 h (n=15).     

Effect of copper on ion- and osmoregulation in the shore crab Carcinus maenas

The uptake and effect of dissolved copper on regulation of hemolymph osmolality and Na⁺, K⁺, Cl^-, Ca⁺⁺, and Mg⁺⁺ concentrations in the shore crab Carcinus maenas (L.) were determined at 400 mOsm (=14 S) ambient salinity. One mg Cu l^-1 resulted in 50% mortality in 11 to 22 d; the highest sensitivity was observed around the moulting period. 0.25 and 0.5 mg Cu l^-1 were not lethal after a onemonth exposure. Ten, 1, and 0.5 mg Cu l^-1 altered regulation of osmolality, Na⁺, K⁺, and Cl^- concentrations, while 0.25 mg Cu l^-1 did not. Exposure to 1 mg Cu l^-1 reduced hemolymph osmolality and Na⁺, K⁺, and Cl^- concentrations to 80 to 90% of controls within 4 to 6 d and no further reduction was observed during a 21-d exposure. The effect of various copper concentrations on these four parameters were almost identical and the highest sensitivity was observed around the moulting period. Hemolymph calcium levels increased 20 to 80% in crabs exposed to 1 mg Cu l^-1, while they decreased 20 to 30% in crabs exposed to 0.5 mg Cu l^-1. Prolonged exposure to copper caused 20 to 70% reductions in hemolymph magnesium levels. Crabs exposed to 0.5 mg Cu l^-1 for 29 d accumulated copper in hepatopancreas, gills, carapace, heart, testes, and hypodermis, but not in muscles and hemolymph. Increased copper levels in crabs exposed to 0.25 mg Cu l^-1 were observed in hepatopancreas, gills, and carapace only. The present results suggest that effects of copper on ion regulatory processes in part explain why the toxicity of copper towards euryhaline invertebrates increases at low salinities.     

Extensive cross-disciplinary analysis of biological and chemical control of Calanus finmarchicus reproduction during an aldehyde forming diatom bloom in mesocosms

Egg and faecal pellet production and egg hatching success of the calanoid copepod Calanus finmarchicus were monitored over a period of 14?days (14?C28 April, 2008) while fed water from 4 differently treated mesocosms and ambient water. Two of the mesocosms used were inoculated with the polyunsaturated aldehyde (PUA)-producing diatom Skeletonema marinoi, while 2 received only nutrient additions with or without silica. The mesocosms developed blooms of S. marinoi, mixed diatoms or the haptophyte Phaeocystis pouchetii, respectively. Faecal pellet production of C. finmarchicus increased with increasing food availability. Egg production increased with time in all mesocosms to a maximum single female production of 232 eggs female^?1?day^?1 (average of 90 eggs female^?1?day^?1) and followed the development of ciliates and P. pouchetii, but was not affected by the observed high (up to 15?nmol?L^?1) PUA production potential of the phytoplankton. The hatching success of the eggs produced on the mesocosm diets was high (78?C96%) and was not affected by either aldehydes in the maternal diet or exposure to the dissolved aldehydes in the water.     

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     

Branchial and intestinal osmoregulatory acclimation in the four-eyed sleeper, <Emphasis Type="Italic">Bostrychus sinensis</Emphasis> (Lacepède), exposed to seawater

Bostrychus sinensis is a facultative air breather that inhabits waters of a wide range of salinities. This study aimed to elucidate whether branchial and intestinal osmoregulatory acclimation occurred in B. sinensis transferred from 5‰ water through a progressive increase in salinities to seawater. Our results indicate that B. sinensis acted as a hyperosmotic regulator in 5‰ water, but exhibited hypoosmotic hypoionic regulation in seawater. During short- (1 day) and medium- (10 days) term acclimation to seawater, there were only minor perturbations in plasma osmolality and [Na⁺], which returned to control levels after 45 days of exposure to seawater. Branchial Na⁺/K⁺-ATPase activity was unaffected by 1, 10 or 45 days of exposure to seawater. However, prolonged (45 days) acclimation to seawater led to a significant increase in Na⁺/K⁺-ATPase α-subunit protein abundance. Taken together, these results indicate that there could be changes in the expression of Na⁺/K⁺-ATPase isoforms and/or post-translational modification of Na⁺/K⁺-ATPase in the gills of fish exposed to seawater. Immunofluorescence microscopy revealed that acclimation to seawater for 10 days only resulted in no change in branchial Na⁺/K⁺-ATPase protein expression, but there were increases in protein expression of cystic fibrosis transmembrane regulator (CFTR)-like chloride channel and Na⁺:K⁺:2Cl⁻ cotransporter (NKCC; probably NKCC1). Indeed, NKCC was undetectable in gills of fish kept in 5‰ water by Western blotting, but it became weakly detectable in fish exposed to seawater for 10 days and prominently expressed in fish exposed to seawater for 45 days. Therefore, our results indicate that branchial CFTR-like chloride channel and NKCC1 were the determining factors in the transition between hyperosmotic regulation and hypoosmotic hypoionic regulation in B. sinensis. Furthermore, the intestine of B. sinensis also served as an important osmoregulatory organ, since there were significant increases in both the activity and protein abundance of intestinal Na⁺/K⁺-ATPase in fish acclimated to seawater for 45 days. The effectiveness of branchial and intestinal osmoregulatory acclimation in B. sinensis during seawater acclimation led to only a minor increase in plasma osmolality, and thus resulted in relatively unchanged free amino acid contents in muscle and liver.     

Reduced hypoosmoregulatory ability and alteration in gill chloride cell distribution in mature chum salmon (Oncorhynchus keta) migrating upstream for spawning

Osmoregulatory ability of mature chum salmon (Oncorhynchus keta) during spawning migration was examined by following the changes in gill Na⁺, K⁺-ATPase activity and in the distribution and morphology of chloride cells. Mature chum salmon caught in Otsuchi Bay, northern Honshu Island, Japan, died within 5 d in seawater (SW) in association with a marked increase in plasma osmolality, whereas the fish transferred to fresh water (FW) maintained plasma osmolality efficiently. Gill Na⁺, K⁺-ATPase activity decreased in both SW-maintained and FW-transferred fish. Well-developed chloride cells, identified by immunocytochemical staining specific for Na⁺, K⁺-ATPase, were present mainly in the filament epithelium of immature fish caught in the ocean. In mature fish caught in the bay, however, additional chloride cells were also found in the lamellar epithelium. The number of filament chloride cells decreased markedly in the mature fish both in SW and in FW, whereas the number of lamellar chloride cells was maintained. These results suggest that the loss of hypoosmoregulatory ability in mature chum salmon may be attributable to the decrease in filament chloride cells and associated decrease in gill Na⁺, K⁺-ATPase activity, and also that appearance of lamellar chloride cells may be preparatory to the forthcoming upstream migration. Received: 14 April 1997 / Accepted: 5 May 1997     

Osmotic and chloride regulation in the hemolymph of the tiger prawn Penaeus monodon during molting in various salinities

The effect of molting on osmotic and chloride concentrations in the blood of the prawn Penaeus monodon Fabricius (20±3 g) at various salinities was investigated. Prawns were obtained from ponds in Iloilo, Philippines, in 1984. They were stocked in salinities of 8, 20, 32 and 44, and their hemolymph was sampled during molt (Time 0), and then 0.125, 0.25, 0.5, 1, 2, 4, 6, 10 and 14 d after molting. Prawns during and immediately after molt tended to conform to the environmental osmolality. Subsequent postmolt (0.5 d) stages displayed more divergence from external salinity. The isosmotic point was higher (940±30 mOsm kg^-1) during molt than during intermolt (663±8 mOsm/kg^-1), suggesting different osmotic requirements in early molt. Hyperregulation of hemolymph chloride below 20 S, as well as isoionic point (301±6 mM), were independent of molting stage. At 20 S and above, newly molted (0 to 0.25 d post-molt) individuals tended to conform to the external chloride concentration while intermolt (0.5 d) post-molt individuals did not. Contribution of hemolymph chloride to hemolymph osmolality was greater during intermolt than during ecdysis, suggesting an important role for other negatively charged ions during molt. When molt occurred in 20 S (the test salinity most similar to the isoionic salinity), there was little or no change in hemolymph osmolality or chloride concentration from 0 to 14 d postmolt. At 8, 32 and 44 S, the change from molt to intermolt values in hemolymph osmotic and chloride concentrations was hyperbolic. Non-linear least-squares regression showed that prawns generally achieved intermolt values within 1 d after molting. Prawns at intermolt regulated hemolymph osmolality (620 to 820 mOsm kg^-1) and chloride concentration (300 to 450 mM) at a much narrower range than during molt (520 to 1 170 mOsm kg^-1 and 250 to 520 mM, respectively). Hemolymph osmolality was a more sensitive indicator of physiological response than hemolymph chloride concentration. Distribution and culture of P. monodon might be limited in low salinities by its ability to maintain a hemolymph osmolality 500 mOsm kg^-1 during molt and 600 mOsm kg^-1 in intermolt, and in high salinities by its capacity to reduce the hemolymph osmolality from values at molt to those in intermolt. Osmotic and chloride concentrations in the blood of P. monodon clearly varied with both molt stage and salinity of the medium. Dependence on external factors, however, gradually declined in older molt stages, suggesting a reduction in integument permeability and greater development of ion absorption/secretion mechanisms as the exoskeleton hardened.SEAFDEC Contribution No. 197     

Osmoregulation in fiddler crabs (<Emphasis Type="Italic">Uca</Emphasis>) from temperate Atlantic and Gulf of Mexico coasts of North America

Fiddler crabs (Crustacea: Decapoda; genus Uca) were collected from 16 locations along the temperate Atlantic and Gulf of Mexico coasts of eastern North America for osmoregulation studies. Three species, U. pugilator, U. pugnax and U. minax, were taken from habitats between Cape Cod, MA, and St. Mary's River, Nassau Co., GA Seven species, U. panacea, U. speciosa, U. spinicarpa, U. longisignalis, U. rapax, U. pugilator and U. minax, were collected in habitats between the Ochlochonee River, Liberty Co., and Thompson's Bayou, Escambia Co., along the Gulf coast of northwest Florida. To examine differences in osmoregulatory capabilities among the species, specimens were placed in various concentrations of artificial seawater (0-3450 mOsm) for 5 days. The oligohaline or "freshwater" species, U. minax, U. spinicarpa and U. longisignalis, possess the lowest average hemolymph osmolality. They are unable to control hemolymph osmolality above 2000 mOsm. On the other hand, the euryhaline species, U. speciosa, U. panacea, U. pugilator and U. pugnax, have much higher average hemolymph osmolality. They are able to withstand an osmotic challenge of 2200 mOsm or greater. Among the eight species, U. panacea and U. pugilator are able to osmoregulate across the broadest range of seawater concentrations (0-2800 mOsm). After an examination of the osmoregulation in several populations of U. minax, U. pugilator and U. pugnax, physiological adaptation is apparent in the two former species, but not the latter. Experimental evidence for capacity adaptation was examined in the freshwater species, U. minax. If slowly adapted to hypertonic seawater (1800 mOsm) for 14 days, this species is able to withstand much higher osmotic pressures than unadapted crabs. Although hemolymph isosmotic concentration remains the same, the limits for regulation are extended to higher osmolality. Consequently, as a result of studying osmoregulation in several species of Uca captured at various locations across their geographic range in the temperate zone, a clearer view of water-balance physiology is available for the genus. The Uca spp. are not uniform in osmoregulatory abilities. There is considerable inter- and intraspecific physiological variation associated with the ecological distribution of each species, respectively.     

Effects of temperature,seawater osmolality and season on oxygen consumption and osmoregulation of the amphipod Gammarus oceanicus

Seasonal variations of oxygen consumption rate, haemolymph osmolality and the concentrations of the inorganic ions potassium and sodium in the haemolymph were measured in the littoral amphipod Gammarus oceanicus collected from the Trondheimsfjorden, Norway in 1987. For each season comparisons were made of amphipods acclimated for 1 wk to 0.5, 4.5, 10.0, 15.0 and 20.0°C, in combination with seawater osmolalities of 100, 500 and 1200 mOsm and to the seawater osmolality corresponding to that of the collecting site. The oxygen consumption rate showed a temperature insensitivity when the amphipods were acclimated to low temperatures in winter and high temperatures in summer. Significant differences were found in oxygen consumption between individuals acclimated to various medium osmolalities, possibly indicating higher energy requirements for osmotic and ionic regulation at low seawater osmolalities. Oxygen consumption rate was significantly higher in summer than in other seasons. Haemolymph osmolality and the concentration of the inorganic ions sodium and potassium were not influenced by temperature or season. Determination of haemolymph osmolalities and concentrations of inorganic ions revealed that G. oceanicus is a strong hyper-osmotic and hyper-ionic regulator in dilute seawater. The concentration of potassium in the haemolymph is less influenced by seawater osmolality than haemolymph osmolality and the haemolymph concentration of sodium.     

Toxicity of cupric lon to eggs of the spot Leiostomus xanthurus and the Atlantic silverside Menidia menidia

Experiments were conducted to determine the toxicity of free cupric ion to the eggs of two marine fish: the spot Leiostomus xanthurus, and the Atlantic silverside Menidia menidia. A cupric ion buffer consisting of 5 mM of the chelator, trishydroxymethylamino methane (Tris), and varied concentrations of CuSO₄ was employed to achieve stable cupric ion activities in experimental seawater media. Egg hatch of the silverside appears to be more sensitive to cupric ion than that of the spot. The silverside had 60% inhibition of hatch at a pCu (negative logarithum of cupric ion activity) of 9.4 with complete supporession of hatching at pCu values below 8.2, while the spot had 50% inhibition at a pCu of 9.0 and complete suppression at a pCu of 8.0. Cupric ion was most toxic to the eggs of silverside at or around the time of hatching, whereas a pronounced sensitivity at hatching was not apparent with the eggs of spot. Calculated estimates of the range of cupric ion activity in seawater indicate that natural activity levels may in some instances inhibit the hatching of silverside and spot eggs.     

Effects of fluctuating salinity on the behaviour of the West African blood clam Anadara senilis and on the osmotic pressure and lonic concentrations of the haemolymph

Specimens of the West African blood clam Anadara senilis were exposed to both sinusoidal and abrupt salinity regimes. Measurements showed that haemolymph osmolality and concentrations of Na, K⁺, Ca²⁺ and Mg²⁺ followed the concentrations of the external medium up to the time of shell valve closure. Shell valves closed when the seawater concentration had fallen to about 15.4% S and reopened at a similar salinity. The closure mechanism was effective in preventing excessive haemolymph dilution.     

Major osmolyte changes during oocyte hydration of a clupeocephalan marine benthophil: Atlantic herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>)

The major inorganic and organic osmolytes responsible for hydrating the oocytes during pre-ovulatory meiotic maturation in autumn- and spring-spawning stocks of Atlantic herring are examined. Despite the ovulated eggs of spring-spawning herring being 1.6- to 2-fold larger than the autumn-spawning stock, the GSI (27 ± 3%) and degree of oocyte hydration (70–72% water) were similar. Normalising the data with respect to dry mass revealed that the physiological mechanisms underlying the maturational influx of water were the same for both classes of egg. Cl⁻, K⁺ and P_i together with a small pool of free amino acids (FAA) represented the driving forces for oocyte hydration. K⁺ (autumn and spring) and P_i (spring) maintained their concentrations in the ovulated eggs, while all other ions, including Cl⁻, Na⁺, NH₄ ⁺ and Mg²⁺ were significantly diluted. In contrast the FAA concentration increased during the hydration process. Amongst the inorganic ions, Cl⁻ showed the greatest increase in the ovulated eggs. The FAA content doubled from 1.5 to 3.3% of dry mass during oocyte hydration and accounted for 29% of the calculated ovoplasmic osmolality in the ovulated eggs from both autumn- and spring-spawners. This significant osmotic effect of the small pool of FAA was due to the low water content of the benthic eggs. The differential movement of the inorganic and organic osmolytes that underly oocyte hydration in Atlantic herring are discussed in relation to current models of transmembrane ion flux.     

The importance of free amino acids to the energy metabolism of eggs and larvae of turbot (Scophthalmus maximus)

A study was undertaken to establish the role of free amino acids (FAA) in aerobic energy dissipation in embryos of turbot (Scophthalmus maximus) which contain an oil globule in the egg. Laboratory-reared developing eggs and larvae (15°C, 34 salinity) were measured for oxygen uptake, ammonia excretion, contents of FAA, protein, and ammonium, and volumes of yolksac and oil globule. Newly spawned eggs from different batches contained 55 to 90 nmol egg^–1 of FAA. Resorption of FAA occurred in parallel with the consumption of yolk. Resorption of the oil globule, however, occurred predominantly after hatching and mainly after yolk resorption. The combined data suggest that approximately 70% of the FAA are utilized as an energy substrate, while the rest are polymerized into body proteins. FAA become a significant energy substrate in the early egg stage and account for 100% of the aerobic energy dissipation 2 d after Fertilization then decrease to ca. 60% at the time of hatching. Lipids derived from the oil globule seem to be the main fuel after hatching and account for ca. 90% of the energy dissipation at the onset of first-feeding. Thus, the energetics of fish embryos which contain an oil globule seems to be different from those that depend exclusively on the nutritional reserves of the yolk.     

Calanoid resting eggs in the Baltic Sea: implications for the population dynamics of Acartia bifilosa (Copepoda)

Resting eggs of Acartia spp. (presumably A. bifilosa Giesbr.) at densities between 65 and 125 eggs cm^-2 were found in sediment off the southern coast of Finland and represent the first observations of dormant calanoid eggs in the Baltic Sea. The highest egg densities were found at sediment depths between 4 and 8 cm. The hatching percentage varied between 0 and 90% in the different sediment layers and at different experimental temperatures (13, 16 and 19°C). Some nauplii emerged from the eggs extracted from sediment layers 9 to 10 cm beneath the surface layer and other nauplii emerged from eggs after 82 d of dark storage at 3°C. Light was not needed to trigger hatching; the nauplii were able to hatch in darkness when the eggs were resuspended in filtered seawater. Indirect evidence suggests that the marked spring peak of Acartia spp. nauplii seen in a monitoring data from the years 1973 to 1984 was derived from benthic eggs and not from spawning. The possible mechanisms for induction of hatching and the adaptive benefits of resting egg production for A. bifilosa are discussed.     

Osmotic and ionic concentration of the egg capsule fluid of Crepidula fornicata in relation to embryonic development

Osmotic pressure and major ions (Cl⁻, Na⁺, Mg²⁺, Ca²⁺) of the egg capsule fluid in the slipper limpet Crepidula fornicata were investigated in relation to embryonic development. Calcium permeability of the capsule wall was studied at oviposition, by dipping freshly laid egg capsules in ⁴⁵Ca as a tracer. This study also determined total calcium content of the embryos at different developmental stages. Osmolarity and major ion concentrations in egg capsule fluid were higher than seawater at uncleaved and trochophore stages, and then dropped to the same level as sea water at veliger stage. Concentrations of Cl⁻ and Na⁺ were relatively high at oviposition, peaked at trochophore stage, and finally dropped close to concentrations of seawater at hatching. In contrast, concentrations of Mg²⁺ and Ca²⁺ decreased steadily during capsular development. Radiotracer permeability experiments in freshly laid egg capsules confirmed that the capsule wall is impermeable to this ion at that stage. However, because of the dissolution of the inner layer of the wall during the final part of capsular development, the wall becomes permeable to calcium and probably to the rest of the major ions studied.     

Abiotic conditions in cephalopod (Sepia officinalis) eggs: embryonic development at low pH and high pCO2

Low pO₂ values have been measured in the perivitelline fluids (PVF) of marine animal eggs on several occasions, especially towards the end of development, when embryonic oxygen consumption is at its peak and the egg case acts as a massive barrier to diffusion. Several authors have therefore suggested that oxygen availability is the key factor leading to hatching. However, there have been no measurements of PVF pCO₂ so far. This is surprising, as elevated pCO₂ could also constitute a major abiotic stressor for the developing embryo. As a first attempt to fill this gap in knowledge, we measured pO₂, pCO₂ and pH in the PVF of late cephalopod (Sepia officinalis) eggs. We found linear relationships between embryo wet mass and pO₂, pCO₂ and pH. pO₂ declined from >12 kPa to less than 5 kPa, while pCO₂ increased from 0.13 to 0.41 kPa. In the absence of active accumulation of bicarbonate in the PVF, pH decreased from 7.7 to 7.2. Our study supports the idea that oxygen becomes limiting in cephalopod eggs towards the end of development; however, pCO₂ and pH shift to levels that have caused significant physiological disturbances in other marine ectothermic animals. Future research needs to address the physiological adaptations that enable the embryo to cope with the adverse abiotic conditions in their egg environment. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Wave action: the environmental trigger for hatching in the California grunion Leuresthes tenuis (Teleostei: Atherinopsidae)

 California grunion Leuresthes tenuis (Teleostei: Atherinopsidae) emerge from the ocean to spawn on beaches of southern California. Grunion eggs do not hatch at a set developmental age, but remain in the sand until turbulent surf at high tide washes them out to sea. In previously studied fishes and amphibians that lay eggs terrestrially, low oxygen is the trigger for hatching in water, and high oxygen tensions inhibit hatching. For the grunion, however, eggs placed in air, seawater and oil did not hatch at any at oxygen tension until they were agitated in fluid. Following agitation in seawater, all eggs hatched within several minutes. Grunion eggs in normoxic or hyperoxic water hatched significantly faster than eggs agitated at the lowest oxygen tensions. Mechanical agitation, not hypoxia, is the environmental trigger for hatching in California grunion eggs. Received: 15 May 1999 / Accepted: 5 April 2000     

Egg production of the Arctic copepod Calanus glacialis: laboratory experiments

Egg production of single female Calanus glacialis Jaschnov fed with Thalassiosira antarctica at super-abundant concentrations (>300 g C l^-1) was determined over several weeks. Experiments were performed directly after collection from the East Greeland Current in June 1987 and 1988, and during resumed feeding after long-term starvation over 4 (October 1988), 4.5 (October 1987) and 6.5 (January 1988) mo. In addition, in June 1987, short-term starvation experiments of 5 and 15 d were conducted. Egg production was closely related to feeding in all experiments. While directly after collection eggs were produced within a few days, it took 2 wk (October 1987 and 1988) and 10 d (January 1988), respectively, to resume egg production after long-term starvation. During long-term starvation periods eggs were not laid. The decrease in total egg production with duration of starvation was due to decreasing clutch size and increasing spawning interval. In contrast, short-term starvation experiments only affected spawning interval. Interannual variability in egg production was high, with much higher clutch sizes in 1988. Average production rates in June 1988 correponded to 5% body C female^-1 d^-1, the maximum was 7.4% (1 274 eggs in 23 d). Carbon content of eggs was 0.40 g egg^-1. C. glacialis is well adapted to pulsed food events in the Arctic by its longevity; its ability to preserve its reproductive potential over several months; its rapid mobilization of ovaries; and by its high egg production rates. The implication of prolonged spawning capacity on life cycle studies is discussed.     

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