Sperm motility and longevity in the giant cuttlefish, Sepia apama (Mollusca: Cephalopoda)

The sperm kinetics and fertilisation literature in marine invertebrates is heavily biased toward free-spawning species. Nonetheless, many species (e.g. cephalopods) transfer and/or fertilise gametes in confined external spaces or internally, creating very different selective pressures on sperm storage, sperm longevity and hence sperm competition. Here we report the results of an investigation into the effects of sperm age, water temperature and sperm concentration on sperm motility in the giant cuttlefish (Sepia apama). Significant positive correlations were found between percent motility and sperm concentration, and between sperm motile speed and sperm concentration. Mean percent motility of cuttlefish sperm suspension was still 9% eight hours after being released from the spermatophore and diluted into filtered seawater at 12°C (ambient field temperature during the spawning season). Sperm resuspended from spermatangia taken from (mated) females in the field were motile for up to 100 hours. When spermatophores were stored at 4°C motility was still observed in resuspended sperm after two months. Our results show that spermatangia and spermatophores can retain and release live sperm for long periods. The observed longevity of sperm in S. apama greatly increases the potential for sperm competition in this species.     

Food effects on statolith composition of the common cuttlefish (Sepia officinalis)

The concentration of trace elements within cephalopod statoliths can provide a record of the environmental characteristics at the time of calcification. To reconstruct accurately the environmental characteristics at the time of calcification, it is important to understand the influence of as many factors as possible. To test the hypothesis that the elemental composition of cuttlefish statoliths could be influenced by diet, juvenile Sepia officinalis were fed either shrimp Crangon sp. or fish Clupea harengus under equal temperature and salinity regimes in laboratory experiments. Element concentrations in different regions of the statoliths (core–lateral dome–rostrum) were determined using laser ablation inductively coupled plasma mass spectrometry (LA- ICPMS). The ratios of Sr/Ca, Ba/Ca, Mn/Ca and Y/Ca in the statolith’s lateral dome of shrimp-fed cuttlefish were significantly higher than in the statolith’s lateral dome of fish-fed cuttlefish. Moreover, significant differences between statolith regions were found for all analysed elements. The fact that diet adds a considerable variation especially to Sr/Ca and Ba/Ca must be taken into account in future micro-chemical statolith studies targeting cephalopod’s life history.     

Respiration of Sepia officinalis during embryonic and early juvenile life

Adult Sepia officinalis L. were caught in June 1984, in the coastal waters of Wimereux (France). Deposition of the eggs took place in the seawater aquaria of the Station Marine. The oxygen consumption of S. officinalis was measured during embryonic and juvenile development. Aerobic metabolism occurs as soon as the early embryonic Stage 21. Oxygen diffuses through the initially thick egg shell; the oxygen level in the perivitelline liquid reaches a maximal value just before hatching (116.7±6.9 mm Hg). Hatchings display only a slight increase in oxygen consumption compared to embryos in the last stage of development. Respiration experiments with 40 d old juveniles showed that oxygen consumption increases with temperature, but is not affected by photoperiod. Experiments under increasing hypoxia revealed that S. officinalis juveniles are good regulators and maintain a constant oxygen consumption in the range of 4 to 7 mg O₂l^-1. Juveniles successfully recover from an hypoxic stress of 2 mg O₂l^-1 maintained for 1 h. This suggests that the respiratory pigments (pre-hemocyanins) of 40 d-old juveniles have a high oxygen affinity and/or that these juveniles have the ability to adapt to anaerobic conditions.     

Behavioral aspects of sperm competition in cuttlefish, Sepia officinalis (Sepioidea: Cephalopoda)

Sexual selection studies in cephalopods indicate that sperm competition is a central feature of their mating systems, yet this has not been studied experimentally in any detail. In 1998 we staged 20 matings of the cuttlefish, Sepia officinalis L., in the laboratory. Males rapidly initiated mating in the “head-to-head” position, with no apparent courtship. Mating lasted an average of 10 min (range 7 to 14 min). For the first 6 min (on average 63% of the mating duration), the male flushed strong jets of water directly at the female's buccal membrane, which sometimes resulted in the expulsion of parts of spermatangia placed there in recent matings. Then, in a single discrete movement that lasted an average of only 14 s, the male's modified fourth left arm – the hectocotylus – wrapped around a single large bundle of spermatophores (ca. 150 to 300) and transferred them to the female's buccal membrane. For the remainder of the mating (average 3 min, range 1.5 to 5.0 min), the hectocotylus repeatedly broke the spermatophores open, and manipulated them, so that sperm were released and many spermatangia were attached along the ventral buccal membrane, near the paired seminal receptacles. Approximately 140 spermatangia were attached in rows 3 to 5 deep around the ventral buccal membrane in a single mating; the rest were usually discarded during mating. Histology revealed that each of the seminal receptacles consists of a series of sperm storage bulbs connected by a central duct, which leads to a single pore at the surface of the buccal membrane. Baseline data on sperm motility were obtained, but the mechanism by which sperm enter the seminal receptacle remains unknown. Females seemed to initiate termination of mating, then males guarded their mates temporarily. These results, combined with other recent laboratory experiments, provide evidence that sperm competition may be a major feature of the mating system of S. officinalis. Received: 4 January 1999 / Accepted: 18 May 1999     

Ocean acidification and temperature rise: effects on calcification during early development of the cuttlefish Sepia officinalis

This study investigated the effects of seawater pH (i.e., 8.10, 7.85 and 7.60) and temperature (16 and 19 °C) on (a) the abiotic conditions in the fluid surrounding the embryo (viz. the perivitelline fluid), (b) growth, development and (c) cuttlebone calcification of embryonic and juvenile stages of the cephalopod Sepia officinalis. Egg swelling increased in response to acidification or warming, leading to an increase in egg surface while the interactive effects suggested a limited plasticity of the swelling modulation. Embryos experienced elevated pCO₂ conditions in the perivitelline fluid (>3-fold higher pCO₂ than that of ambient seawater), rendering the medium under-saturated even under ambient conditions. The growth of both embryos and juveniles was unaffected by pH, whereas ⁴⁵Ca incorporation in cuttlebone increased significantly with decreasing pH at both temperatures. This phenomenon of hypercalcification is limited to only a number of animals but does not guarantee functional performance and calls for better mechanistic understanding of calcification processes.     

The effect of ration size, temperature and body weight on specific dynamic action of the common cuttlefish Sepia officinalis

The effect of meal size (shrimp Crangon crangon) [0.83–18.82% dry body weight (Dw)] on specific dynamic action (SDA) was assessed in cuttlefish Sepia officinalis (1.03–6.25 g Dw) held at 15 and 20°C. Cuttlefish <2 g significantly expended less energy in feeding and digesting their meal than cuttlefish >2 g when given the same quantity of food. Handling, eating and digesting a shrimp meal was temperature dependent with cuttlefish processing and digesting a similar sized shrimp meal faster at 20°C than at 15°C. The proportional increase in oxygen consumption (2.07 ± 0.02) was not correlated with feeding rate (FR) and was independent of temperature and cuttlefish size. The SDA peak was not correlated with FRs, and increased as cuttlefish size and temperature increased. The mean SDA coefficient was 0.87 ± 0.07% of the ingested energy; one of the lowest SDA values recorded amongst vertebrates and invertebrates. Daily energy requirements (KJ day⁻¹) for S. officinalis were calculated from laboratory estimates of energy losses due to standard (MO_{2 Standard}), routine (MO_{2 Routine}) and feeding (MO_{2 SDA}) oxygen consumption. Laboratory estimates of daily metabolic expenditures were combined with results from previous investigations to construct an energy budget for 1 and 5 g cuttlefish consuming a meal of 5 and 15% Dw at 20°C and the amount of energy available for growth was estimated to be between 35 and 80.3% of the ingested energy.     

Respiration and nitrogen metabolism of Atlantic halibut eggs (Hippoglossus hippoglossus)

Naturally spawned and fertilized eggs of Atlantic halibut,Hippoglossus hippoglossus L., were analysed for protein, free amino acids (FAA), ammonium ions and energy content. The chemical composition was found to be size-dependent but varied little during egg development. Ammonium ions did, however, accumulate during the late embryonic stage, and the trend in FAA content was downward during the same period. Rates of O₂ uptake and NH₃ excretion followed exponential patterns. A total of 1µmol O₂ was consumed and 120 nmol NH₃ excreted between the time intervals of fertilization and 1 d post hatch. Derived O:N ratios indicated that the dominant portion of the energy metabolism was lipid- or carbohydrate-based during the mid-development period but switched to FAA as hatch was approached.Correspondence should be addressed to: Zoological Laboratory, University of Bergen, Allégt. 41, N-5007 Bergen, Norway     

Influence of temperature and salinity on the trace element incorporation into statoliths of the common cuttlefish (<Emphasis Type="Italic">Sepia officinalis</Emphasis>)

The use of statolith chemistry to trace migration pathways and distinguish populations of cephalopods is based on the assumption that the elemental composition of statoliths is influenced by physicochemical properties of the ambient environment. However, such influences have not been investigated experimentally up until now. This study presents the first microchemical analyses of cephalopod statoliths obtained from laboratory experiments under different controlled temperature and salinity conditions. Our results show that statolith chemical composition is strongly related to both salinity and temperature in ambient waters. The Ba/Ca ratio is negatively related to temperature and shows no relation to salinity. The I/Ca ratio is positively related to temperature and negatively to salinity. No Sr/Ca relation was found to either salinity or temperature, suggesting that the well-established proxy strontium is not as useful in cephalopod statoliths as in other biomineralized aragonites. Microanalysis of trace elements, however, shows an enormous potential for field studies on distribution, migration and stock separation of cephalopods. Furthermore, Synchrotron X-ray Fluorescence Analysis is introduced as a promising novel method for statolith analysis, providing a spatial resolution of typically 10–15 μm combined with detection limits down to 0.5 ppm.     

Effect of body mass,temperature and food deprivation on oxygen consumption rate of common cuttlefish <Emphasis Type="Italic">Sepia officinalis</Emphasis>

Predictions of short and long term changes in Sepia officinalis metabolism are useful, since this species is both economically important for aquaculture and also is an ideal experimental laboratory organism. In this study standard and routine oxygen consumption rates of newly hatched and juvenile laboratory raised cuttlefish S. officinalis ranging between 0.04 and 18.48 g dry body mass (Dm), were measured over a range of temperatures (10, 15, 20 and 25°C). The mass exponent (b) ranged between 0.706 and 0.992 for standard oxygen consumption and between 0.694 and 0.990 for routine oxygen consumption. Oxygen consumption scaled allometrically (b = 0.7) with body mass for cuttlefish <2 g Dm and isometrically (b = 1) thereafter. No significant differences were apparent amongst the slopes of oxygen consumption and body mass at different temperatures for standard and routine oxygen consumption. However, the intercepts differed significantly amongst the regression lines, indicating a significant effect of temperature on the magnitude of oxygen consumption. The combined effect of temperature (T) and dry body mass (Dm) are best described by the following equations: cuttlefish <2 g, MO₂ = 0.116Dm^0.7111.086 ^T and >2 g, MO₂ = 0.076Dm^0.9831.091 ^T for standard oxygen consumption; cuttlefish <2 g, MO₂ = 0.538Dm^0.7291.057 ^T and >2 g, MO₂ = 0.225Dm^0.9621.081 ^T for routine oxygen consumption. Using these equations it was estimated that a cuttlefish of 1 g Dm held at 20°C, eating 5% Dm day⁻¹ and undergoing standard and routine metabolism consumes 21.3 and 35.4%, respectively of its total daily energy intake. Juvenile cuttlefish (3.32–5.08 g Dm) held at 15°C and deprived of food for 27 days maintained a stable standard oxygen consumption rate for the first 6 days following starvation. By the 18th day without food, oxygen consumption rate had declined by 53% and further declined to 65% below the standard oxygen consumption rate on the 27th day. Upon resumption of feeding, the respiration rate returned immediately to the initial level prior to food deprivation. The present study defines the basic energy requirements and general physiological state of young cuttlefish at temperatures of 10–25°C with and without food.     

Lipid composition of eggs and adductor muscle in giant scallops (Placopecten magellanicus) from different habitats

Scallops (Placopecten magellanicus Gmelin) were collected during August 1989 from shallow water (10 m) and deep water (31 m) habitats at Sunnyside, Trinity Bay, Newfoundland, to compare the lipid composition of eggs and adductor muscle tissue. Less favorable food levels and lower temperature conditions associated with deeper water have previously been shown to produce slower growth and reduced fecundity in individuals from this habitat. Triacylglycerol reserves consistently accounted for 60% of the total lipids present in both groups. The total lipid content of the eggs and the composition of their triacylglycerol fatty acid pools were similar in shallow water and deep water scallops, indicating very little if any nutritional difference between the two groups. Relative to their counterparts from shallow water, individuals from deeper water contained higher proportions of docosahexaenoic acid [22:6(n-3)] in the egg phospholipids and higher levels of 24-methylenecholesterol (a phytosterol commonly found in diatoms) in the adductor muscle. Differences in fatty acid composition are interpreted as biochemical adjustments of cell membranes to increase membrane fluidity, thereby compensating for the lower temperatures prevailing at the greater depth.     

Respiration of the bathypelagic fish Cyclothone acclinidens

The respiration of 3 adult females of Cyclothone acclinidens was measured in situ. These numerically dominant fish of the meso- and bathypelagic zones were individually captured off southern California at a depth of 1300 m using a multiple-chambered slurp-gun respirometer operated from the submersible Alvin. Continuous oxygen consumption measurements of each fish were made at the capture depth for about 28 h, spanning an interval from late morning to the following mid-afternoon. Nocturnal rates of respiration of these fish were 3 to 5 times higher than diurnal rates, suggesting a daily thythm in this non-migratory midwater species. Environmental factors such as tides, light, and food could serve to synchronize the observed respiration fluctuation.     

Respiration and excretion by the ctenophore Mnepiopsis leidyi

Respiration (dissolved oxygen and carbon dioxide) and excretion (dissolved organic carbon, inorganic and organic nitrogen and phosphorus) rates were measured for a variety of sizes of Mnemiopsis leidyi over a temperature range of 10.3° to 24.5°C. Both respiration and excretion rates were a direct linear function of animal weight and very temperature sensitive (Q₁₀4). Oxygen uptake ranged from 155 to 489 g at O/(g dry weight) day^-1 and carbon dioxide release from 43 to 166 M. Organic carbon made up about 38% of the total carbon released. Inorganic nitrogen excretion, exclusively in the form of ammonium, comprised 54% of the total nitrogen release and ranged from 10 to 36 M NH₄/(g dry weight) day^-1. Average release of dissolved primary amines (expressed as glycine equivalents) equaled 43% of the organic nitrogen fraction. Inorganic phosphorus release ranged from 2.0 to 4.9 M/(g dry weight) day^-1 and made up about 72% of the total phosphorus loss. The turnover of elements in the body was calculated as 5 to 19% per day for carbon and nitrogen, depending on the temperature, and an even higher 20 to 48% per day for phosphorus. These values are comparable to rates observed for small, active zooplankton.     

Determination of copper in embryos and very young specimens of Sepia officinalis

The total amount of copper in embryos and newly hatched young individuals of Sepia officinalis L. has been determined by microtechnique, using bathocuproine-sulfonate as complexing reagent. During embryonic life, the total amount of copper does not change; it remains at a level close to 3.8 g. The copper is found in the yolk sac of very early embryos; it is subsequently transferred into the embryo proper. After hatching, the copper content diminishes quickly in starved individuals. Fed S. officinalis also usually lose copper. The reason for this may be that the inner yolk sac of newly hatched individuals contains a great deal of the total copper, which is excreted with the yolk after the latter has become superfluous. Later on, copper must be taken up from the food. The mobilization of protein and copper from the yolk into the blood may account for the early appearance of embryonic hemocyanin in the blood.     

Respiration and chemical composition of the bathypelagic euphausiid Bentheuphausia amblyops

Bentheuphausia amblyops is a cosmopolitan bathypelagic euphausiid with a vertical range of from 280 to 7 000 m. Determinations of proximate and elemental composition show that B. amblyops has a more robust structure (lower water content and higher protein content) than is typical of other bathypelagic Crustacea. B. amblyops is a strong swimmer and is capable of employing its thoracic legs in raptorial fashion. Discrete depth trawls taken between depths of 400 and 2 500 m on a diel basis show no evidence of vertical migration. There was no significant difference in oxygen consumption within the species environmental temperature range (1.5° to 7.5°C), which may be partially ascribed to a limited temperature effect and partially to variability in spontaneous activity at the different temperatures. Between 7.5° and 9.5°C there was a marked temperature effect on metabolism (Q₁₀=6.4). The observed vertical distribution and metabolic response to elevated temperature preclude the migrations to the surface that have been postulated for B. amblyops. The robust composition of the species, its behavioral characteristics, and published diet information strongly suggest that the species is omnivorous with a strong predatory component in its foraging strategy.     

Sodium- and potassium-activated adenosine triphosphatase in the excretory organs of Sepia officinalis (Cephalopoda)

Sodium- and potassium-activated ATPase (Na⁺–K⁺-ATPase) has been demonstrated in excretory organs of Sepia officinalis, using a cytochemical procedure. In the renal appendages, both epithelia of the pancreatic appendages, the folded epithelium of the branchial heart appendage and the transport-active epithelium of the gill, the enzyme is localized exclusively in the basolateral cell membranes, i.e., the membranes of the basal labyrinth and the lateral plasma membranes. In addition, Na⁺–K⁺-ATPase is also located in the sarcolemma of the muscle fibres of the branchial heart. Distribution and localization of the enzyme is further substantiated by [³H]-ouabain autoradiography. The possible involvement of Na⁺–K⁺-ATPase in the excretion of ammonia and in ionic regulation in dibranchiate cephalopods is discussed.This study was supported by the Deutsche Forschungsgemeinschaft and is part of a doctoral dissertation     

Respiration and nitrogen excretion by the squid Loligo forbesi

The amphipod Corophium volutator (Pallas) of the Bay of Fundy is a major food resource for migrating shorebirds. A relatively low total lipid content (ca 1.7%) was found in summer and fall samples of 1986. Analysis of triacylglycerides which would normally reflect an animal diet through fatty acid input, revealed high proportions of fatty acids characteristic of algae. C. volutator contains hydrocarbons (0.2% of total lipids) with a unimodal distribution in the range C₁₈ to C₃₅ with maximum at C₂₅, and a weak odd-carbon preference (carbon preference index, C.P.I. =1.22). This hydrocarbon pattern shown by C. volutator seems to be biogenic, but due to both a weak odd-carbon preference and a significant content of the isoprenoid phytane, the presence of petroleum hydrocarbons cannot be ruled out. Comparison of the hydrocarbon composition be ruled out. Comparison of the hydrocarbon composition found in C. volutator with hydrocarbons described in the literature for its most likely sources of food suggests that highly degraded material and aged detritus are components of the diet of this deposit feeder, and as important as the local diatom population. No organohalogen pesticides (DDT, dichlorodiphenyl-trichloroethane, or derivatives) or polychlorinated biphenyls (PCBs) were detected in the lipid extracts of C. volutator. Absence of these materials and low hydrocarbon levels explain the continued successful exploitation of this food resource by migrating shorebirds.     

Cuttlebone calcification increases during exposure to elevated seawater pCO2 in the cephalopod Sepia officinalis

Changes in seawater carbonate chemistry that accompany ongoing ocean acidification have been found to affect calcification processes in many marine invertebrates. In contrast to the response of most invertebrates, calcification rates increase in the cephalopod Sepia officinalis during long-term exposure to elevated seawater pCO₂. The present trial investigated structural changes in the cuttlebones of S. officinalis calcified during 6 weeks of exposure to 615 Pa CO₂. Cuttlebone mass increased sevenfold over the course of the growth trail, reaching a mean value of 0.71 ± 0.15 g. Depending on cuttlefish size (mantle lengths 44–56 mm), cuttlebones of CO₂-incubated individuals accreted 22–55% more CaCO₃ compared to controls at 64 Pa CO₂. However, the height of the CO₂-exposed cuttlebones was reduced. A decrease in spacing of the cuttlebone lamellae, from 384 ± 26 to 195 ± 38 μm, accounted for the height reduction The greater CaCO₃ content of the CO₂-incubated cuttlebones can be attributed to an increase in thickness of the lamellar and pillar walls. Particularly, pillar thickness increased from 2.6 ± 0.6 to 4.9 ± 2.2 μm. Interestingly, the incorporation of non-acid-soluble organic matrix (chitin) in the cuttlebones of CO₂-exposed individuals was reduced by 30% on average. The apparent robustness of calcification processes in S. officinalis, and other powerful ion regulators such as decapod cructaceans, during exposure to elevated pCO₂ is predicated to be closely connected to the increased extracellular [HCO₃ ⁻] maintained by these organisms to compensate extracellular pH. The potential negative impact of increased calcification in the cuttlebone of S. officinalis is discussed with regard to its function as a lightweight and highly porous buoyancy regulation device. Further studies working with lower seawater pCO₂ values are necessary to evaluate if the observed phenomenon is of ecological relevance.     

Osmotic properties of eggs of the herring Clupea harengus

Yolk osmolality of developing eggs of the herring Clupea harengus L. is strongly hypoosmotic to seawater: about 440 mOsm during the first week of development decreasing to 360 mOsm before hatching. The perivitelline fluid (PVF) of the eggs is isoosmotic to the ambient medium. The PVF equilibrates within 10 min to changes in the ambient seawater. The content of Na⁺, K⁺, Cl^- and free amino acids amount to 26, 52, 48, and 54 n mol egg^-1, respectively, on Days 1 to 3 after fertilization, increasing to 63, 69, 80, and 79 n mol egg^-1, respectively, prior to hatching (Days 18–20). The apparently conflicting findings of a decreasing yolk osmolality and a simultaneous increase in the amount of egg solutes are at present unexplained.     

A density-gradient column for determining the specific gravity of fish eggs,with particular reference to eggs of the mackerel Scomber scombrus

A new method is described for determining the specific gravity of fish eggs. The technique requires the preparation of a stable column of a continuously graded solution of sea-water salts such that a specimen introduced into the column settles at a level of known specific gravity, where it is in hydrostatic equilibrium with the fluid in the column. The method allows the simultaneous measurement of specific gravity of a number of specimens continuously throughout development and is suitable for use aboard a vessel at sea. Differences of specific gravity may be resolved to a precision of better than 4x10^-5; as an example of use of the method, the specific gravity of eggs of the mackerel Scomber scombrus L. declined during development within the range 1.0205 to 1.0245; unfertilized eggs displayed a continuous increase of specific gravity.