Distribution of lipids between the zooxanthellae and animal compartment in the symbiotic sea anemoneAnemonia viridis: Wax esters,triglycerides and fatty acids

The temperate sea anemoneAnemonia viridis (Forskäl) contained about 11% lipid on a dry weight basis when maintained at light levels of about 10µE m^–2 s^–1 and a temperature of 10°C. Aposymbiotic forms of the anemone had similar lipid levels. These values are very low compared with tropical symbiotic Anthozoa in which lipid levels constitute up to 50% of dry weight. In symbioticA. viridis, <6% of total lipid consisted of the storage lipids, wax esters and triglycerides. Most of the triglyceride was stored in the animal tissues rather than the zooxanthellae. Zooxanthellae contained only small amounts of wax esters. An analysis was made of the wax ester, triglyceride and fatty acid composition of symbiotic anemones, isolated zooxanthellae and aposymbiotic anemones. Wax ester composition was similar in symbiotic and aposymbiotic forms. However, triglyceride composition differed. In particular trimyristin (C42) was found only within the symbiotic association. Fatty acids showed a high degree of unsaturation, and acids with both even and odd numbers of carbon atoms were found. The most abundant fatty acid was 16:0 in all samples, except for the total lipids from zooxanthellae in which the major fatty acid wastrans-18:1.     

Effect of light on the total lipid content and storage lipids of the symbiotic sea anemoneAnemonia viridis

In order to examine the effect of light level on the storage lipids of the symbiotic sea anemoneAnemonia virudis (Forskäl), anemones were exposed to three experimental light regimes of 10, 100 and 300 E m^-2s^-1. Anemones were fed once a week. After 30 d there were no significant differences in the total lipid levels between anemones at any of the light intensities. However, after 60 d lipids had increased in proportion to light level in both the animal-tissue and zooxanthellae compartments. The higher levels of total lipid were in part due to increases in storage lipid (wax esters and triglycerides). Wax ester levels increased in the animal tissues but remained constant in the zooxanthellae, whereas triglycerides increased in both compartments. In contrast to fed anemones, starved anemones which were maintained at 300 E m^-2s^-1 for 30 or 60 d did not show a statistically significant change in lipid levels at 60 d, although a slight increase in the lipid level was observed. However, there was a significant increase in the storage lipids, which suggested that the non-storage phospholipids and structural lipids had declined as a result of cellular catabolism. The composition of the wax esters and triglycerides of both fed and starved anemones was analysed and compositional changes were observed at higher light intensities.     

Structure of and energy reserves in the liver of wild and cultured yellowtail flounder,<Emphasis Type="Italic"> Limanda ferruginea</Emphasis>

Knowledge of the structure and energy reserves, in the liver of commercially important fish species, is important in understanding metabolic processes and in assessing the impact of potential environmental physical and chemical stressors in both wild and cultured stocks. The present study investigated the microscopic morphology and histochemistry (total and neutral lipids, glycogen) of liver tissue of wild (3 ⁺) and cultured (1 ⁺) sexually immature female and male yellowtail flounder ( Limanda ferruginea Storer), sampled in late April 2001. Hepatosomatic indices [HSI: (liver weight/body weight-liver weight)×100] of cultured fish were significantly higher than those of wild fish. Females in the cultured group had significantly lower HSIs than males. The liver of both wild and cultured L. ferruginea was interspersed with pancreatic tissue. The main components of the liver tissue were irregular cords of hepatocytes arranged in tubules which surrounded vascular sinusoids. The hepatocytes contained an abundance of lipid, much of which appeared to be neutral lipids, in both sexes of the cultured fish. Total and neutral lipid droplets were larger, and the area occupied by these droplets was significantly greater in both cultured females and males compared to the wild fish, suggesting lipidosis in the cultured fish. In the cultured fish these differences were sex-dependent, the females having significantly more total and neutral lipids in hepatocytes than the males. This suggests a potentially greater storage capacity in females and/or a higher lipid metabolism in males. There were no statistically significant differences in glycogen content between the cultured and wild fish, or between the sexes in both sampling groups.     

Physiological energetics of the intertidal sea anemone Anthopleura elegantissima

Energy budgets were calculated for individuals of the sea anemone Anthopleura elegantissima (Brandt), collected in 1981 and 1982 from Bodega Harbor, California, USA. Rates of ammonium excretion were measured in high-and low-intertidal, symbiotic and aposymbiotic sea anemones within 24 h of collection. Among symbiotic and aposymbiotic individuals, no differences in excretion rate were found on the basis of intertidal height. However, rates of ammonium excretion in aposymbiotic anemones (2.14 mol NH ₊ ⁴ g^-1 h^-1) were significantly higher than in symbiotic ones (0.288 mol NH ₊ ⁴ g^-1 h^-1). Rates of excretion were used with estimated rates of oxygen uptake to calculate nitrogen quotients (NQ). NQ and RQ values from the literature were used to calculate an oxyenthalpic equivalent [501 kJ (mol O₂)^-1 for R+U], and mass proportions of protein (54%), carbohydrate (44%) and lipid (2%) catabolized during routine metabolism in this species 24 h after feeding. Integrated energy budgets of these experimental anemones were calculated from data on ingestion, absorption and growth, and estimates of translocated energy from the symbiotic algae. Contribution of zooxanthellae to animal respiration based on translocation=90% and RQ=0.97 are 41 and 79% in high-and low-intertidal anemones, respectively. Calculated scope for growth is greater than directly measured growth in both high-and low-intertidal individuals. The deficit, estimated as 30% of assimilated energy in high-intertidal anemones, is attributed to unmeasured costs (specific dynamic effect) or production (mucus). Low-intertidal anemones lost mass during the experiment, implying that the magnitude of the deficit was greater in these anemones than in upper intertidal individuals. Anemones from both shore levels lost zooxanthellae during the experiment, which contributed to energy loss since the contribution of the zooxanthellae is greater in low-intertidal anemones. Scope for growth is preserved in high-intertidal anemones (29% of assimilated energy) because metabolic demands are lower due to aerial exposure, and prey capture rate is higher compared to lowshore anemones. Although possibly underestimated, lower scope for growth in low-shore anemones may result from continuous feeding and digestion processes that are less efficient than those of periodically feeding high-intertidal anemones.     

Effect of feeding regime and irradiance on the photophysiology of the symbiotic sea anemone Aiptasia pulchella

To determine how the animal and algal components of the symbiotic sea anemone Aiptasia pulchella respond to changes in food availability and culture irradiance, sea anemones from a single clone were maintained at four irradiance levels (320, 185, 115, and 45 E m^-2 s^-1) and either starved or fed for 5 wk. Changes in protein biomass of sea anemones maintained under these conditions were not related to the productivity of zooxanthellae, since the protein biomass of fed A. pulchella decreased with increase in irradiance and there was no difference in protein biomass among starved sea anemones at the four irradiance levels. Except for the starved high-light sea anemones, the density of symbiotic zooxanthellae was independent of culture irradiance within both starved and fed. A. pulchella. Starved sea anemones contained over twice the density of zooxanthellae as fed sea anemones. Within both starved and fed individuals, chlorophyll per zooxanthella increased with decreasing culture irradiance while algal size remained constant (in fed sea anemones) at about 8.80 m diameter. Chlorophyll a: c ₂ ratios of zooxanthellae increased with decreasing culture irradiance in zooxanthellae from starved sea anemones but remained constant in zooxanthellae from fed sea anemones. As estimated from mitotic index data, the in situ growth rates of zooxanthellae averaged 0.007 d^-1 and did not vary with irradiance or feeding regime. Photosynthesis-irradiance (P-I) responses of fed A. pulchella indicated an increase in photosynthetic efficiency with decreasing culture irradiance. But there was no consistent pattern in photosynthetic capacity with culture irradiance. Respiration rates of fed sea anemones also did not vary in relation to culture irradiance. The parameter I _k , defined as the irradiance at which light-saturated rates of photosynthesis are first attained, was the only parameter from the P-I curves which increased linearly with increasing culture irradiance. The daily ratio of net photosynthesis to respiration for A. pulchella ranged from 1.6 to 2.8 for sea anemones maintained at the three higher irradiances, but was negative for those maintained at 45 E m^-2 s^-1. Since the final protein biomass was greatest for sea anemones maintained at the lowest irradiance, these results indicate that sea anemone growth cannot be directly related to productivity of zooxanthellae in this symbiotic association.     

Isolation,characterization and glucose metabolism of glycogen cells (=vesicular connective-tissue cells) from the labial palps of the marine mussel Mytilus edulis

A method has been developed to isolate glycogen cells (=vesicular connective tissue cells) from the labial palps of the marine mussel Mytilus edulis L. These cells have a modal density of 1.14 g ml^-1 and this property has been exploited to separate them from pronase-dissociated cell dispersions by density-gradient centrifugation. The isolated cells were mainly spherical, with a small peripheral nucleus. The quasitotality of the cell was filled with -like glycogen particles with cellular organelles being limited to the cell periphery. The major recipients of [U-¹⁴C] glucose-derived carbon were glycogen and amino acids. Only small quantities of ¹⁴C radiolabel were recovered from lipids, protein and CO₂. Glucose incorporation into glycogen increased linearly over 6 h and showed saturation kinetics with respect to exogenous glucose concentration.     

Nutritional exchange in a tropical tripartite symbiosis: direct evidence for the transfer of nutrients from anemonefish to host anemone and zooxanthellae

The relationship between anemones and anemonefishes is an oft-cited and endearing example of a mutualistic symbiosis. Current research on mutualistic symbioses suggests these relationships are more commonplace and have greater importance at the ecosystem level on nutrient dynamics and evolutionary processes than previously thought. Using stable isotopes ¹⁵N and ¹³C, both field and laboratory experiments were designed to investigate whether nutrient transfer from two species of resident anemonefishes (Amphiprion perideraion and A. clarkii) to host anemones (Heteractis crispa) occurs. Mass spectroscopy indicated that both ¹⁵N and ¹³C were significantly elevated in the tissues of anemonefishes and in both host anemone and zooxanthellae fractions. These experiments provide the first direct empirical evidence of nitrogen and carbon transfer from resident anemonefishes to host anemones and endosymbiotic zooxanthellae. Such transfer of elements within this intriguing tripartite association underscores the central role that nutrient dynamics contributes to the evolutionary processes of these marine symbioses.     

A comparative analysis of the photobiology of zooxanthellae and zoochlorellae symbiotic with the temperate clonal anemone <Emphasis Type="Italic">Anthopleura elegantissima</Emphasis> (Brandt). III. Seasonal effects of natural light and temperature on photosynthesis and respiration

The sea anemone Anthopleura elegantissima hosts two phylogenetically different symbiotic microalgae, a dinoflagellate Symbiodinium (zooxanthellae, ZX) and a chlorophyte (zoochlorellae, ZC). The photosynthetic productivity (P), respiration (R), and contribution of algal carbon translocated to the host (CZAR) in response to a year’s seasonal ambient changes of natural light and temperature are documented for both ZX- and ZC-bearing anemones. Light and temperature both affect photosynthesis, respiration, and CZAR, as well as various algal parameters; while there are evident seasonal differences, for the most part the relative effects on P, R, and CZAR by the two environmental variables cannot be determined. Net photosynthesis (P_n) of both ZX and ZC was significantly higher during spring and summer. During these seasons, the P_n of ZX was always greater than that of ZC. Regardless of algal symbiont, anemone respiration (R) was significantly higher during the spring and summer. The annual net carbon fixation rate of anemones with ZX and ZC was 325 and 276 mg C anemone⁻¹ year⁻¹, respectively, which translates to annual net community productivity rates of 92 and 60 g C m⁻¹ year⁻¹ for anemones with ZX or ZC, respectively. CZAR did not show a clear relationship with season; however the CZAR for ZX was always significantly greater than for ZC. Lower ZX growth rates, coupled with higher photosynthetic rates and higher CZAR estimates, compared to ZC, suggest that if A. elegantissima is simply carbon limited, ZX-bearing anemones should be the dominant symbiont in the field. However ZC-bearing anemones persist in low light and reduced temperature microhabitats, therefore more than the translocation of carbon from ZC must be involved. Given that global climate change will increase water temperatures, the potential for latitudinal range shifts of both ZC and ZX (S. californium and muscatinei) might be used as biological indicators of thermal shifts in the littoral zone of the Pacific Northwest.     

Physiological energetics of the intertidal sea anemone Anthopleura elegantissima

High-intertidal (H) individuals of the sea anemone Anthopleura elegantissima (Brandt) are exposed aerially up to 18 h each day, unlike low-intertidal (L) individuals which may be continuously immersed over many days. Thus, H anemones experience shorter feeding periods compared to L anemones. From 1980 to 1982, H and L anemones were observed and collected at the mouth of Bodega Harbor in North Central California (USA) to determine whether any physiological adaptations mitigate the energetic effects of reduced feeding time in H anemones. Weight of prey in coelenterons of H anemones was three times more than that of L anemones following a single immersion period. This difference is not due to slower digestion rates in H anemones. Prey residence time in coelenterons (4h) was equivalent in both groups. Different prey weights imply that ingestion rates were greater in H individuals. However, all anemones had similar weight-specific feeding-surface areas. Different prey-capture rates result from increased receptivity to prey in H anemones, rather than from increases in feeding surface. Absorption efficiency was inversely related to ration size in anemones from both shore positions. H individuals absorbed food more efficiently than L individuals fed equivalent rations. Ration, not exposure conditions, affected absorption efficiency. Daily growth rates were 1.5 to 1.8% and 1.2 to 1.4% of dry body weight in H and L anemones fed large rations (4.0 to 5.6% of dry body weight), respectively. H anemones fed smaller daily rations, approximating amounts of zooplankton captured naturally (1% of anemone dry weight), had higher growth rates and growth efficiencies than L anemones, which lost mass. Higher growth rates in H anemones, which are supported by higher prey-capture rates, result in attainment of minimum body size for reproduction in a relatively short period of time despite reduction in time available for feeding, thus improving relative fitness of these anemones in the upper intertidal zone.     

Changes in the biochemical composition of the ovaries of the seastar Asterias rubens during its annual reproductive cycle

The biochemical composition of the ovaries of the asteroid echinoderm Asterias rubens was studied, with emphasis on the changes which are manifest during the annual reproductive cycle. Ovarian dry weight displays only minor changes during ovarian development, constituting about 20% of the organ fresh weight. Levels of all constituents show some variation, but in general increase with ovarian growth. Only the free amino acid level is rather constant. Except for glycogen, the contents of other constituents (total lipids, other reducing sugars, free amino acids and proteins) show a rectilinear increase with ovarian growth. Glycogen content increases rectilinearly with time during early vitellogenesis, but remains constant during further development until ovarian maturity. By plotting constituent levels and contents against a non-linear gonad index axis rather than against sampling date, results can be correlated more easily with ovarian growth stages. The possibility of endocytosis of a vitellogenin-like yolk precursor by the ovary is discussed.     

Biochemical composition and trophic strategies of the amphipod Eurythenes gryllus at hadal depths (Atacama Trench,South Pacific)

Amphipods Eurythenes gryllus were collected at 7800?m depth in the Atacama Trench (South Pacific) for studying their biochemical composition (in terms of proteins, lipids and carbohydrates and fatty acid content) and to gather information on bioenergetic strategies and trophic habits of organisms living in this extreme environment. The effect of long-term formalin storage on the biochemical determinations was also determined. Proteins were the dominant biochemical class of organic compounds (39–53%D.W.), whereas carbohydrates accounted for a very small fraction (1–2%D.W.). Lipid concentrations of E. gryllus accounted for 7–18%D.W. and were much lower than those reported for other deep-sea amphipods. These differences are likely to be more dependent upon food availability in the Atacama Trench rather than upon temperature. Lipid composition of E. gryllus revealed the dominance of monounsaturated fatty acids with polyunsaturated fatty acids accounting for a very small fraction, suggesting that hadal amphipods are higher dependent upon lipid reserves than amphipods inhabiting at shallow depths. The ratio of C18:1Δ⁹ to C18:1Δ¹¹ was >11 confirming the necrophagous trophic habits of this hadal amphipod.     

Changes in the biochemical composition of the pyloric caeca of female seastars,Asterias rubens,during their annual reproductive cycle

The biochemical composition of the pyloric caeca of female seastars (Asterias rubens) was studied throughout the annual reproductive cycle. The pyloric caeca index is high during the pseudo-rest stage of the ovaries and decreases gradually during ovarian growth. The pyloric caeca dry weight varies between 20 and 38% of the fresh weight. Lipids, and less prominently carbohydrates (both glycogen and other reducing sugars), seem to constitute the primary nutrient reserves for gonad growth. Proteins are also available to meet ovarian requirements during vitellogenesis. Glycogen levels are high in late autumn and winter and low at spawning time and in summer, whereas levels of the other reducing sugars are high in post-spawning months and subsequently gradually decrease during gonad development. The total lipid level is extremely variable (averaging 75 mg g^-1 organ fresh weight), peak levels of over 200 mg g^-1 being found in some individuals in summer and at spawning time. The free amino acid level is constant at about 20 mg g^-1. The protein level is rather high throughout the annual cycle, and displays only minor changes. The observed patterns of decrease in major constituents suggest a rapid release of nutrient reserves from the pyloric caeca (deposited during the summer) at the onset of vitellogenesis. Moreover, the needs of the ovaries during further development seem to be so tremendous that during the winter months the digested food may be transported to the ovaries without prior deposition in the pyloric caeca. The changes observed in the pyloric caeca are discussed and compared with those occurring simultaneously in the ovaries.     

Copper uptake by the sea anemoneAnemonia viridis and the role of zooxanthellae in metal regulation

Anemonia viridis (Forskäl) were collected from south-west Scotland and south-west England in October 1988. When exposed to 0.05 and 0.2mg 1^–1 copper in sea water, anemones did not take up the metal in proportion to external concentrations. Results suggested thatA. viridis regulated copper by expelling symbiotic algae (or zooxanthellae) which were shown to accumulate copper. The use of aposymbiotic (non-zooxanthellate) anemones in similar metal-uptake experiments indicated that other mechanisms may also be involved in metal regulation. Mucus was produced byA. viridis when the anemone was exposed to copper, and it is proposed that mucus may be involved in the regulation process. The implication of this work on the use of coelenterates as biological indicators of environmental metal levels is discussed.     

Symbiont photosynthesis increases both respiration and photosynthesis in the symbiotic sea anemone Anemonia viridis

Dark respiration of the symbiotic sea anemone Anemonia viridis (Forskäl) was observed to increase by 34% when anemones were exposed to hyperoxic sea water (150% oxygen saturation) overnight, and by 39% after exposure to 6 h in the light at a saturating irradiance of 300 E m^-2 s^-1 at normoxia (100% oxygen saturation). No increase due to light stimulation was observed in aposymbiotic control anemones. In darkness, the oxygen concentration of the coelenteric fluid was hypoxic. However, within 10 min of anemones being illuminated, coelenteric fluid was hyperoxic, and it remained elevated throughout a 12 h light period. When measured over a 24 h period (12 h light: 12 h dark), the dark respiration rate increased gradually over the first 6 h of the light period until it was 35% above the dark night-time resting rate. It remained elevated throughout the remaining light period and for 2 h into the following dark period, after which it fell back to the resting rate. Gross photosynthesis (P ^gross) increased significantly when anemones were exposed to either hyperoxia (150% oxygen saturation) or 300 E m^-2 s^-1 at normoxia. This increase was not observed when symbiotic anemones were illuminated at a low-light intensity of 100 E m^-2 s^-1. The results of this study suggest that respiration in the dark is limited by oxygen diffusion and that normal respiration is restored in the daytime by utilisation of the oxygen released by photosynthesis. Furthermore, it appears that the increased respiration following exposure to high-light intensities provides a CO₂-rich intracellular environment which further enhances the photosynthetic rate of the zooxanthellae.     

Host feeding and nutrient sufficiency for zooxanthellae in the sea anemone Aiptasia pallida

Nutrient sufficiency of zooxanthellae in the sea anemone Aiptasia pallida cultured in low nutrient seawater depends on the availability of particulate food to the host. Zooxanthellae in anemones unfed for 20 to 30 d exhibited the following characteristics of nutrient deficiency: cell division rates decreased; chlorophyll a content gradually decreased from 2 to <1 pg cell^–1; and C:N ratios increased from 7.5 to 16. Over a 3-mo period, algal populations in unfed anemones gradually decreased, indicating that zooxanthellae were lost faster than they were replaced by division. The mitotic index of zooxanthellae in unfed anemones was stimulated either by feeding the host or by the addition of inorganic N and P to the medium. Whether algae are nutrient-limited in hosts under field conditions has not been examined fully; however, C:N ratios in zooxanthellae from field-collected hosts are slightly higher (9.4 vs 7.5) than in hosts fed to repletion in laboratory cultures. This observation might indicate N limitation in the field.     

Effects of starvation,and light and dark on the energy metabolism of symbiotic and aposymbiotic sea anemones,Anthopleura elegantissima

Rates of oxygen and carbon-dioxide exhange were measured in symbiotic and aposymbiotic specimens of the sea anemone Anthopleura elegantissima while fed and starved under light or dark conditions. Respiratory quotients indicated that fed anemones switched from a carbohydrate to a fat catabolism when starved, with the exception that symbiotic individuals starved in the light showed a pronounced carbohydrate catabolism for over 1 month. The source of the carbohydrate was probably photosynthate translocated by the dinoflagellate Symbiodinium (=Gymnodinium) microadriaticum (Freudenthal) living in the anemones' tissues. The starved symbiotic anemones maintained in the light had lipid levels not significantly different from fed controls and 44 to 61% higher than starved aposymbiotic anemones after 1 month. Thus, the quality and quantity of the metabolic flux from the symbionts to the sea anemone were sufficient to conserve the host's lipid reserves.     

Chemische tarnung. Die stoffliche grundlage der anpassung von anemonenfischen an riffanemonen

Dissolved amino acids (³H-phenylalanine, ³H-proline) were accumulated and incorporated into proteins and glycoproteins by sea anemones. On objects (glass rods, pipe cleaners or filter paper) which had touched labelled anemones, tritiated substances could be detected. The mucus of labelled anemones was analysed by disc-electrophoresis. Tritium activity was found in the bands (stained by amino black). Anemone fishes adapted to labelled anemones had 4 times more tritium activity on their surface than control fishes. Most radioactivity was found on those regions which made closest contact with the anemones. The mucus of anemone fishes adapted to labelled anemones was also analysed by disc-electrophoresis. The analysis revealed protein-containing fractions in which ³H-amino acids were incorporated. The pattern of tritium activity in disc-electropherograms from labelled anemones corresponds to that of fishes adapted to those anemones. Anemones produce specific substances which influence the discharge of their nematocytes. These substances have special functions in the normal behaviour of the anemones. The substances provide protection against self-nettling and prevent the discharge of nematocytes into nearby objects which the tentacles continuously contact. The production of these substances by anemones is completely separate from the association with anemone fishes. The fishes thus simply exploit a mechanism existing independently in the anemones. Therefore, it is possible to form, in an aquarium, unnatural associations between anemone fishes and anemone species which never live in association with fishes in their natural biotope. On the basis of previous information, as well as on these new data, it is possible to develop a model which explains the control of nematocyte discharge: Substances with inhibitory qualities (protecting substances) are produced by the anemones themselves, and de-sensitize the sensory inputs of the anemones (nematocytes and sensory cells). Sensitization takes place as soon as the anemones come into contact with “stimulating substances”. This happens if anemones are touched by food objects or by anemone fishes which have been previously isolated from anemones. The surfaces of these fishes are not impregnated with protecting substances. Adapted anemone fishes, neighbouring anemones of the same species and other “adapted” objects are coated with the inhibitory substances and thus do not induce nematocyte discharge.     

Nutrient transfer in a marine mutualism: patterns of ammonia excretion by anemonefish and uptake by giant sea anemones

Many symbioses involve multiple partners in complex, multi-level associations, yet little is known concerning patterns of nutrient transfer in multi-level marine mutualisms. We used the anemonefish symbiosis as a model system to create a balance sheet for nitrogen production and transfer within a three-way symbiotic system. We quantified diel patterns in excretion of ammonia by anemonefish and subsequent absorption by host sea anemones and zooxanthellae under laboratory conditions. Rates of ammonia excretion by the anemonefish Amphiprion bicinctus varied from a high of 1.84 μmole g⁻¹ h⁻¹ at 2 h after feeding, to a basal rate of 0.50 μmole g⁻¹ h⁻¹ at 24–36 h since the last meal. Conversely, host sea anemones Entacmaea quadricolor absorbed ammonia at a rate of 0.10 μmole g⁻¹ h⁻¹ during the daytime in ammonia-enriched seawater, but during the night reduced their absorption rate to near zero, indicating that ammonia uptake was driven by zooxanthella photosynthesis. When incubated together, net ammonia excretion was virturally zero, indicating that host anemones absorbed most of the ammonia produced by resident fish. Adult anemonefish weighed about 11 g under laboratory conditions, but on the coral reef may reach up to 64 g, resulting in a maximal potential ammonia load of >200 μmole h⁻¹ produced by two adult fish during daylight hours. In contrast, host sea anemones weighed about 47 g in the laboratory, but under field conditions, large individuals may reach 680 g, so their maximal ammonia clearance rates may reach about 70 μmole h⁻¹ during the daytime. As such, the ammonia load produced by adult anemonefish far exceeds the clearance rate of host anemones and zooxanthellae. Ammonia transfer likely occurs mainly during the daytime, when anemonefish consume zooplankton and excrete rapidly, and in turn the zooxanthellae are photosynthetically active and drive rapid ammonia uptake. We conclude that zooplanktivorous fishes that form mutualisms with coral reef cnidarians may serve as an important link between open water and benthic ecosystems, through the transfer of large quantities of nutrients to zooxanthellate hosts, thus enhancing coral reef productivity.     

Observations on the biochemical changes in gonads and other organs of Uca annulipes,Portunus pelagicus and Metapenaeus affinis (Decapoda: Crustacea) during the reproductive cycle

The fluctuations in biochemical constituents such as water, nitrogen, non-protein nitrogen, protein, lipid and glycogen in gonad, muscle and hepatopancreas have been followed in 3 decapod crustaceans, Uca annulipes (Latreille), Portunus pelagicus (Linnaeus) and Metapenaeus affinis (Milne-Edwards). The water and ash content of the entire body show no systematic fluctuation in relation to the annual reproductive cycle. The water content of the ovary diminishes as it matures. In the ovary of these crustaceans, the lipid fluctuated greatly in relation to the reproductive cycle. The maturing ovary contains more lipid than an immature or spent ovary per unit tissue weight. The changes in the biochemical constituents in the testis are not so pronounced as in the ovary, since the testicular cycle is often drawn-out and almost continuous in these crustaceans. There is an inverse relationship between water content and lipid content of the hepatopancreas; the greater the fat content, the lesser the water content. The hepatopancreas in these crustaceans is apparently a storage organ and contains much lipid and glycogen. At the height of the breeding season, when gonad production is intense, there is an indication of the mobilisation of at least a part of the lipid from hepatopancreas to gonad.Dissertation for the award of a Doctorate Degree of the Kerala University.Formerly the Marine Biological Laboratory.     

Seasonal variations in biochemical constituents during the reproductive cycle of the female dog cockle Glycymeris glycymeris

In an attempt to describe the biochemical events associated with the main stages of the annual and reproductive cycles of the female dog cockle Glycymeris glycymeris L., we studied seasonal variations in the various stages of oocyte development of the ovaries, and the glycogen, total protein and total lipid content of five body tissues – adductor muscle, foot, tunic coat, visceral mass and mantle. From November 1991 to November 1994, microscopic examination of the ovaries and measurement of the tissue concentrations of glycogen, total proteins and total lipids in these five body tissues were made monthly on ten female dog cockles originating from the sea area around Douarnenez (south Brittany, France). Morphological studies revealed that in the population investigated the annual cycle is characterised by three major periods: a first period of vitellogenesis extending from February/March to April/May and preceding a spawning in spring; a second period of vitellogenesis extending from May/June to September/October and leading to either no spawning, a single autumnal spawning event, or to two spawning events in summer and autumn; and a third period extending from October/November to February/March and characterised by a high level of oocyte lysis. In the muscular body tissues of the dog cockle, i.e. the adductor muscles, the foot and the tunic coat (the muscular envelope containing the visceral mass), the concentrations of glycogen, total proteins and total lipids underwent very similar variations during the annual cycle. During each stage of vitellogenesis, a typical glycogen–protein–lipid sequence was observed in the muscular tissues that was characterised firstly by a peak of glycogen concentration 2 to 3 mo before spawning, followed by a peak in total proteins 1 mo before spawning, and finally by a peak in lipid content just before spawning. A similar glycogen–protein–lipid sequence was also recorded in the first half of the winter period. However, these events were followed by general atresia affecting all oocytes in the gonad. Maximum energetic value of biochemical constituents in females coincided with peaks in lipid content in the visceral mass and mantle. These biochemical events occurred principally immediately before and at the end of oocyte lysis (December/January). A drop in the total energetic value, affecting mainly the visceral mass and the mantle, was recorded each year during the period January to March, coinciding with the period of shell growth in this species. Our data clearly indicate that in female G. glycymeris all muscular tissues contribute to the storage of glycogen and proteins, and suggest that glycogen may be the source of energy triggering vitellogenesis. Biochemical and microscopic observations revealed that oocyte development takes place during the first half of winter, but that these oocytes undergo atresia in December/January. The metabolites produced from oocyte lysis could contribute to somatic growth, which occurs in late winter. Received: 3 March 1997 / Accepted: 23 July 1997