Testing for the Effects of Seasonal and Lunar Periodicity on the Reproduction of the Edible Sea Urchin <Emphasis Type="Italic">Tripneustes gratilla</Emphasis> (L) in Kenyan Coral Reef Lagoons

The annual and lunar reproductive cycle of the widely distributed edible sea urchin Tripneustes gratilla (L) was examined through measurements of gonad index, histological examination of gametogenesis, and induction of spawning with KCl injections. The population density and morphological characteristics of urchins at Diani, Kanamai, and Vipingo reef lagoons were also studied as well as the effects of seawater temperature and light on reproduction. Gonad growth started early during the northeast monsoon and reached a peak in June at the beginning of the southeast monsoon followed by a sharp decrease in gonad size of 50% in July and August towards the end of the southeast monsoons. Histological examination of gonads, revealed many different stages of gametogenesis with gametes present throughout the year, indicating continuous reproduction. There was a significant relationship between gonad index and lunar day with spawning occurring between lunar day 7 and 21, but spawning was not in perfect synchrony in the population. The population density of urchins at each reef is variable from year to year and was highest on average at Vipingo. Urchins at Kanamai had the lowest gonad indices, the largest jaws and smallest individuals an indication of food limitation. The gonads (roe) of T. gratilla at all three sites, were perpetually ‘runny’ an attribute that is not suitable for urchin fisheries. Studies to develop techniques to improve roe quality are recommended.     

How diet influences energy partitioning in the regular echinoid Psammechinus miliaris; constructing an energy budget

The effect of diet (made from either animal or plant material) on the partitioning of energy in small (8-16 mm test diameter (td)) and large (29-37 mm td) Psammechinus miliaris was examined. Diet significantly affected ingestion, digestion and growth of both size groups. Assimilation rates of the different types of feed varied and they supported differential development of the body parts. Urchins fed on an algal diet showed poorer absorption efficiency, assimilation, gonadal and somatic growth than individuals fed on an artificial sea urchin diet (mixed plant and animal material) or on the animal based diets. Small urchins fed on a diet of salmon food utilised proteins as the primary energy source rather than carbohydrates or lipids as indicated by low O/N atomic ratios, and salmon diet promoted a higher energetic investment in reproductive development in both size classes. A negative energy balance resulted when large urchins were fed on the algal diet. Energy losses due to ammonia excretion were negligible and the metabolic losses of assimilated energy, measured as oxygen consumption, were between 0.2% and 1.5% with the different diets. A diet made of mussel flesh stimulated ingestion, gave the highest assimilation rates and best overall growth performance. Such detailed information should assist in the design of diets for the cultivation of sea urchins.     

The effect of dietary protein and carbohydrate concentration on the biochemical composition and gametogenic condition of the sea urchin Lytechinus variegatus

Previously starved urchins, Lytechinus variegatus, (36.0 ± 0.8 (SE) mm test diameter) were held in replicated (3) 10-L aquaria with artificial seawater at 22 ± 2  °C and 32‰ salinity and fed three diet treatments. Urchins were fed diets containing 9 : 35, 20 : 23 or 31 : 12% dry protein: % dry carbohydrate (P : C) ad libitum for a 65-day period. Gonads from urchins fed the 9 : 35 P : C diet had similar organic, lower ash, and lower water content than urchins fed the 31 : 12 P : C protein diet. Water content varied with both diet and nutritional history; consequently, water content may have limited value as a predictor of gonad nutritional status. Protein and carbohydrate concentrations in the gonad were directly related to the dietary composition of these nutrients; gonad lipids did not vary with diet. Excess carbohydrates are frequently stored as fats in fish and mammals but this does not appear to be the case in L. variegatus. Test carbohydrate storage and gut protein storage also reflected dietary composition. Image analysis of ovaries indicated decreased nutritive phagocyte volume, increased germinal epithelium volume and larger oocyte diameters in urchins fed high protein, low carbohydrate diets. Analysis of testes also indicated decreased nutritive phagocyte volume and increased gamete volume with urchins fed high protein, low carbohydrate diets, but differences among treatments were less obvious than in ovaries. This study suggests that high protein, low carbohydrate diets promote gamete growth and development. In addition, the biochemical and gametic composition of gonads can be altered by manipulating dietary composition. This could affect the quality and value of sea urchin roe for human consumption.     

Select light spectra affect gonad color in the sea urchin Lytechinus variegatus

It is suggested that gonad color in sea urchins depends upon the in vivo accumulation and metabolism of red and yellow carotenoid pigments. We hypothesized that differential light exposure could affect carotenoid deposition and, hence, gonad color in sea urchins. We therefore performed two experiments to determine whether light spectra affect the gonad color of Lytechinus variegatus. In the first experiment, urchins were fed a formulated feed supplemented with or without β-carotene and held beneath three lighting regimes designed to emit differing wavelengths of the visible spectrum. After 12 weeks, urchins were dissected and gonad color (CIE L*a*b*) was measured with a Pantone Capsure RM200. Actinic light significantly increased the value of a* (red) in gonad color. Color in the orange and yellow spectra in the gonads increased in individuals fed the β-carotene supplemented diet. In the second experiment, we cultured urchins for nine weeks under lamps specialized to emit UV radiation. All urchins in this experiment received diets supplemented with β-carotene. There was no significant difference in harvested gonad color between these treatments. These data suggest that light quality and dietary carotenoids affect carotenoid deposition in the gonads.     

Extraocular vision in the sea urchin Diadema setosum

In most organisms, specific structures are responsible for detecting light patterns and vision. Several species of sea urchins appear to have a diffuse photoreceptor system that enables them to detect light and in some cases objects. The presence of extraocular vision was investigated on a sea urchin common in northern Oman, Diadema setosum. Urchins from 8- to 10-m deep coral communities were used in controlled experiments that quantified the orientation response of the urchins to various visual cues: size, contrast and colour of circular targets simulating urchins’ outlines. Urchins responded to black discs down to 11° initial visual angle but not to smaller discs. The minimum grey-scale value triggering a response was between 50 and 37% black. Urchins responded to red targets but not to green or blue ones. An average angular distance between spines of 5.3° suggested a visual acuity of about 10.6°. D. setosum thus showed extraocular vision allowing them to detect objects of different sizes, contrasts and to some extent colour, supporting the hypothesis that the whole sea urchin’s body acts as a single large compound eye.     

Carotenoids in the sea urchin Paracentrotus lividus: occurrence of 9'-cis-echinenone as the dominant carotenoid in gonad colour determination

Regular sampling of wild Paracentrotus lividus was carried out over a 12-month period to examine seasonal effects on the pigment profile and content of the gonads, especially in comparison to gonad colour. The major pigments detected in the gut wall were breakdown products of fucoxanthin, namely fucoxanthinol and amarouciaxanthin A. Lower levels of other dietary carotenoids (lutein and β-carotene) together with some carotenoids not found in the diet, namely isozeaxanthin and echinenone ( 20% total carotenoid) were also detected in the gut wall. The presence of echinenone in the gut wall demonstrates that this organ acts as a major site of carotenoid metabolism. Echinenone is the dominant carotenoid in the gonads, accounting for approx. 50–60% of the total pigment. Both all-trans and 9′-cis forms of echinenone were detected in both the gut wall and in the gonad, with levels of the 9′-cis form typically 10-fold greater than the all-trans form in the gonad. The detection of large levels of 9′-cis-echinenone in wild sea urchins is unexpected due to the absence of 9- or 9′-cis forms of carotenoids in the natural, algal, diet. Whilst echinenone clearly contributes towards gonad pigmentation, levels of this carotenoid, cannot be directly linked to a qualitative assessment of gonad colour in terms of market acceptability. Indeed, unacceptable gonad colouration can be seen with both very low and high levels of echinenone and total carotenoid. The presence of 9′-cis-echinenone as the major carotenoid contributing to the pigmentation/colour of the gonad is an important observation in terms of developing artificial diets for urchin cultivation.     

Changes in growth and reproduction of green sea urchins, Strongylocentrotus droebachiensis (Müller), during repopulation of the shallow subtidal zone after mass mortality

Following a disease outbreak that caused mass mortality of green sea urchins, Strongylocentrotus droebachiensis, along the Atlantic coast of Nova Scotia in September 1999, changes in growth and reproduction were monitored over 3.75 years as surviving individuals migrated from deep water to repopulate the shallow subtidal zone at a wave-exposed site. Urchins were sampled at 4 depth strata: at 24 m on a boulder field where the population was unaffected by the disease, at 12 and 16 m on a steeply sloping bedrock ramp, and at 8-10 m along the lower margin of a kelp bed (Laminaria digitata) where urchins formed a grazing front by January 2002. Urchins migrating shoreward from the deep-water refuge responded rapidly to increased algal productivity in the shallows through increased growth and reproduction. Measures of annual increments of skeletal elements (rotules) from urchins across the depth gradient indicated that the fastest growing individuals from the source population formed the grazing front. Urchins in the front reached a larger asymptotic size and produced larger gonads than urchins lower on the ramp. The annual cycle in gonad index showed a pronounced spring spawning period across all depths; a secondary fall spawning was evident at the front and 12 m. The presence of mature, fertilizable ova and short response time to spawning induction in both spring and fall supported the occurrence of two spawning periods.     

Effect of macrophyte diet and initial size on the survival and somatic growth of sub-adult <Emphasis Type="Italic">Strongylocentrotus purpuratus</Emphasis>: a laboratory experimental approach

Recently recruited urchins from the same brood, but with different initial sizes, may respond differently to similar environmental factors. The aim of this study was to assess and compare the effects of starvation and diet on the survival, growth rates in size and weight, and gonad index among small and large sub-adult purple sea urchins, Strongylocentrotus purpuratus. Small urchins ranged from 7.3 to 7.8 mm and large urchins from 11.8 to 14.1 mm (test diameters). Two independent experiments were performed. In the first experiment, sea urchins were fed during 22 weeks on Egregia menziesii (ad libitum) and for only 1 day month⁻¹ (starved condition). Feeding regime significantly affected survival, somatic growth rate in size and weight, and gonad index, with higher means in the ad libitum treatments than in starving conditions. A recurrent cannibalism event by conspecifics occurred in small sea urchins under starving conditions. In the second experiment, sea urchins were fed during 13 weeks ad libitum with four diets: kelp (E. menziesii), coralline algae (Bossiella orbigniana), eelgrass (Phyllospadix scouleri) and a mixed diet of the three species. Survival was not affected by diet or urchin size, but diet significantly affected somatic growth rate in size and weight and gonad index. Kelp promoted the highest growth rate (2.23 ± 0.21 mm month⁻¹), the mixed diet produced an intermediate growth (1.26 ± 0.21 mm month⁻¹), while the lowest values corresponded to coralline algae and the eelgrass (0.30 ± 0.12 and 0.10 ± 0.03 mm month⁻¹, respectively, means ± SE). The mean growth rate of small urchins (averaging all diets) was higher than in large specimens (1.17 ± 0.37 and 0.77 ± 0.28 mm month⁻¹, respectively).     

On the dynamics of the mass mortality of Diadema antillarum in Barbados

Widespread mortality of the black sea urchin Diadema antillarum occurred in the Caribbean in 1983; beginning in Panama in January, and having its major impact at Barbados in September. Mortality on ten reefs surveyed in Barbados was 93.2%, with the highest being 99.9% and the lowest 86.9%. Mortality on each reef was independent of the pre-mortality density on the reef. Urchins with test diameters between 20 and 40 mm were more severely affected than smaller or larger urchins. Populations on reefs exposed to incoming oceanic water suffered heavier mortality than those on protected reefs. Mean size of urchins was smallest on high density reefs. This may indicate a negative effect of density on urchin growth. At post-mortality densities, urchins may grow faster and reach sexual maturity sooner.     

Identification of protein components from the mature ovary of the sea urchin Evechinus chloroticus (Echinodermata: Echinoidea)

The gonads of sea urchins are a high value seafood product, with considerable research being undertaken worldwide on the development of sea urchin aquaculture. As the best prices are obtained for specific gonad attributes, research has also focused on the development of artificial diets that enhance gonad quality and quantity. Total protein has been used as a measure of gonad quality; yet no studies to date have applied proteomics technology to diet development. Here we use a MudPIT and 2-DE approach to describe the major proteins in mature ovaries of a New Zealand sea urchin Evechinus chloroticus. This tissue, which is a target seafood product, contained 138 proteins that were identified from the recently completed sea urchin genome (Strongylocentrotus purpuratus) with high confidence. The majority of these proteins had general functions, with only 12 related to ovarian reproductive function. Eighteen proteins were located on the 2-DE; four of these were directly identified from S. purpuratus protein sequences. In combination this paper shows that the genome resources of S. purpuratus can be used to identify proteins in sea urchins from different families; describes the proteome of E. chloroticus mature ovary; and, provides proteomic tools for analysis of gonads from other edible sea urchins.     

Fatty acid compositions of gonadal material and diets of the sea urchin, Psammechinus miliaris: trophic and nutritional implications

The fatty acid compositions of gonadal material was examined for the sea urchin Psammechinus miliaris (Gmelin) held in aquaria and fed either salmon feed pellets or the macroalga, Laminaria saccharina for 18 months. Gonadal material was also examined from P. miliaris collected from four field sites, including commercial scallop lines encrusted with the mussel, Mytilus edulis, sea cages stocked with Atlantic salmon Salmo salar and two intertidal sea-loch sites, characterised by either a fine mud or a macroalgal substratum. The fatty acid compositions of known and potential dietary material was examined. The proportions of certain fatty acids in the gonads of P. miliaris were significantly affected by diet type and location. Docosahexaenoic acid (DHA) 22:6 n-3 was significantly higher in the gonads of the sea urchins fed salmon feed in aquaria and collected from the salmon cages and scallop lines than in the gonads of the sea urchins fed L. saccharina in aquaria and collected from the intertidal sea loch sites. The salmon feed and the mussel tissue also contained a high proportion of this fatty acid. Stearidonic acid 18:4 n-3 and arachidonic acid 20:4 n-6, however, were found in significantly higher proportions than DHA in the gonads of the sea urchins fed L. saccharina and collected from the two intertidal sea-loch sites. L. saccharina was also found to contain high proportions of stearidonic and arachidonic acid. The gonads of the sea urchins collected from the intertidal site, characterised by a mud substratum, and from the scallop lines were found to contain a lower 18:1 n-9/18:1 n-7 ratio and a higher proportion of branched and odd-chained fatty acids, signifying a high dietary bacterial input, than the sea urchins held in the aquaria and collected from the salmon cage. 20:2 and 22:2 non-methylene-interrupted dienoic fatty acids (NMIDs) were found in P. miliaris fed diets lacking these fatty acids suggesting de novo biosynthesis. These results, therefore, suggest that the proportions/ratios of certain fatty acids in the gonads of P. miliaris could be used to give an indication of the predominant diet type of this species in the wild.     

The effect of the predator-avoidance behavior of the sea urchin,Centrostephanus coronatus,on the breadth of its diet

Summary The sea urchin, Centrostephanus coronatus, exhibits a relatively broad diet. Evidence presented in this paper suggests that evolution of dietary breadth is favored by two mechanisms. First, individual urchins occur in habitat types that support somewhat different foods. Urchins consume principally the most preferred food where it is common but are forced to eat other food types where it is rare. Second, overgrazing by individual urchins reduces the local abundance of the most preferred food, thereby forcing them to add less preferred items to their diet. Previous work indicates that the escape behavior of Centrostephanus from its fish predator, Pimelometopon pulchrum, results in different individuals being exposed to different foods and in reduction of the size of urchin grazing ranges to the point that overgrazing occurs. Hence, by the two mechanisms discussed in this paper, the presence of the fish predator favors the broad diet of its sea urchin prey.     

Habitat-dependent growth in a Caribbean sea urchin <Emphasis Type="Italic">Tripneustes ventricosus</Emphasis>: the importance of food type

The sea urchin Tripneustes ventricosus is a common, yet relatively poorly known, grazer of seagrass beds and coral reefs throughout the Caribbean. We compared the size and abundance of urchins between adjacent seagrass and coral reef habitats (where macroalgae are the dominant primary producers). We also conducted a laboratory experiment comparing the growth rate of juvenile urchins fed a diet of either macroalgae or seagrass. Reef urchins had significantly larger test diameter than those in the seagrass on some sampling dates. This size difference may be at least partially explained by diet, because laboratory-reared urchins fed macroalgae grew significantly faster than those fed seagrass. The seagrass population, however, was stable over time, whereas the reef population exhibited strong fluctuations in abundance. Overall, our study indicates that both the seagrass and coral reef habitats are capable of supporting healthy, reproductive populations of T. ventricosus. Each, however, appears to offer a distinct advantage: faster growth on the reef and greater population stability in the seagrass.     

Extraction and analysis of carotenoids from the New Zealand sea urchin Evechinus chloroticus gonads

Sea urchin gonad (roe) is a highly valued food in Japan and North America. Gonad price is strongly influenced by quality, with appearance, especially colour being a major determinant. Previous attempts to extract a carotenoid profile from the New Zealand sea urchin species Evechinus chloroticus have been challenging due to the large amount of lipid present in the gonad. A carotenoid extraction and high performance liquid chromatography (HPLC) analysis method was developed to reduce lipid contamination by incorporating a saponification and lipid cold precipitation in the extraction procedure. This method enabled greater carotenoid purity and enhanced analysis by HPLC. Echinenone was found to be the main carotenoid present in all E. chloroticus gonads. Dark coloured gonads contained higher levels of fucoxanthin/fucoxanthinol, β-carotene and xanthophylls such as astaxanthin and canthaxanthin. This information on the modification and deposition of carotenoids will help in the development of diets to enhance gonad colour.     

Long-Term Changes in the Gonad Condition of the Sea Urchin Strongylocentrotus intermediusfrom Alekseev Bight (Amurskii Bay) under Polluted Conditions

The gonad condition of the sea urchin Strongylocentrotus intermediuscollected in August 1997 at two stations in Peter the Great Bay was examined. One of the stations was located in a polluted area (Alekseev Bight, Popov Island) and the other, in a relatively clean area (the Verkhovskii Islands). The results were compared with analogous data for 1984, 1985, and 1989. In 1997, the gonad condition of sea urchins inhabiting the two areas differed significantly. The mean value of the gonad index (GI) for sea urchins from Alekseev Bight was less than half and the maturity index was about twice that of sea urchins from the Verkhovskii Islands. The GI of sea urchins from Alekseev Bight decreased by a factor of 1.5 between 1984 and 1997. Pronounced histopathological changes were found in sea urchin gonads at this station: granular and hydropic dystrophy of oocytes, resorption and a sharp decrease in the number of gametes (in about 20% of the sea urchins, hardly any gametes were found in the gonads), changes in the morphology of accessory cells (hypertrophy, atrophy, and necrosis), and accumulation of lipofuscin in the cytoplasm of accessory cells and oocytes, in the hemal sinuses and mesentery. The suppressed gonad condition of the sea urchin S. intermediusin Alekseev Bight may be a consequence of the unfavorable environmental situation that formed in the bight in the 1980s–1990s. The main negative factor is anthropogenic pollution of Amurskii Bay.     

Reduced density of the herbivorous urchin Diadema?antillarum inside a Caribbean marine reserve linked to increased predation pressure by fishes

Disease has dramatically reduced populations of the herbivorous urchin Diadema antillarum Philippi on Caribbean reefs, contributing to an increased abundance of macroalgae and reduction of coral cover. Therefore, recovery of D. antillarum populations is critically important, but densities are still low on many reefs. Among the many potential factors limiting these densities, the focus of this study is on predation pressure by fishes. Marine reserves provide opportunities to examine large-scale manipulations of predator–prey interactions and, therefore, D. antillarum densities were compared inside and outside a reserve in The Bahamas (Exuma Cays Land and Sea Park; ECLSP). Urchins and their fish predators were surveyed at nine sites inside and outside the ECLSP. Because of lower fishing effort, the total biomass of urchin predators, weighted by their dietary preferences for urchins, was significantly higher inside the ECLSP. Furthermore, fish community structure was significantly different inside the Park because of the increased biomass of the majority of species. No urchins were seen inside the ECLSP and this was significantly lower than the density of 0.04 urchin m⁻² outside the Park. Regression analysis indicated that the relationship between the biomass of urchin predators and the proportion of transects containing urchins was non-linear, suggesting that small increases in fish biomass dramatically reduce urchin abundances. The link between lower density of urchins and higher density of their predators inside the ECLSP is strengthened by discounting five alternative primary mechanisms (variations in macroalgal cover, larval supply, environmental setting, density of other urchin species and abundance of predators not surveyed). Caribbean marine reserves have an important conservation role, but increased fish predation appears to reduce densities of D. antillarum. Urchins currently have limited functional significance on Bahamian reefs, but any future recovery of D. antillarum is likely to be limited in reserves, with potentially important ecological consequences. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Sutural loosening and skeletal flexibility during growth: determination of drop-like shapes in sea urchins

The shape of sea urchins may be determined mechanically by patterns of force analogous to those that determine the shape of a water droplet. This mechanical analogy implies skeletal flexibility at the time of growth. Although comprised of many rigid calcite plates, sutural collagenous ligaments could confer such flexibility if the sutures between plates loosened and acted as joints at the time of growth. We present experimental evidence of such flexibility associated with weight gain and growth. Over 13-, 4-, and 2-week periods, fed urchins (Strongylocentrotus droebachiensis) gained weight and developed looser sutures than unfed urchins that maintained or lost weight. Further, skeletons of fed urchins force-relaxed more than did those of unfed urchins and urchins with loose sutures force-relaxed more than those with tight sutures. Urchins (Strongylocentrotus franciscanus) fed for two and a half weeks, gained weight, also had looser skeletons and deposited calcite at sutural margins, whereas unfed ones did not. In field populations of S. droebachiensis the percentage having loose sutures varied with urchin diameter and reflected their size-specific growth rate. The association between feeding, weight gain, calcite deposition, force relaxation and sutural looseness supports the hypothesis that urchins deform flexibly while growing, thus determining their drop-like shapes.     

Effect of dietary history and algal traits on feeding rate and food preference in the green sea urchin Strongylocentrotus droebachiensis

Feeding behaviour is influenced by a variety of factors, including nutritional requirements, the quality of available foods, and environmental conditions. We examined the effect of two factors, food morphology and dietary history, on the feeding rate and preference of the sea urchin Strongylocentrotus droebachiensis. Standardizing food shape and structure did not alter urchins' expected preference for the native kelp Laminaria longicruris over the invasive alga Codium fragile ssp. tomentosoides. However, when foods containing L. longicuris were shaped to mimic the algae, the C. fragile mimic was consumed more rapidly than the kelp mimic. Dietary history had no effect on single diet feeding rate. Urchins feeding on C. fragile consistently consumed twice as much (by mass) as those fed kelp, regardless of their previous diet. Despite higher feeding rates on C. fragile, urchins feeding on this alga were unable to compensate for its low energetic content and ingested less energy. Dietary history had a short-term effect on food preference, with urchins tending to prefer less familiar foods. Our findings suggest that urchins feed on C. fragile at a high rate, due to ease of handling and/or compensatory feeding, and that they do not a have strict preference hierarchy. Rather, food choice appears to reflect active maintenance of a mixed diet.     

The effect of feeding or starvation on resource allocation to body components during the reproductive cycle of the sea urchin Sphaerechinus granularis (Lamarck)

To determine the effects of feeding or starvation on resource allocation to body components during the reproductive cycle of Sphaerechinus granularis, sea urchins were placed in laboratory tanks and either fed ad libitum or starved during two different periods of their biological cycle, i.e. the mature stage and the recovery stage. The urchin growth was monitored over the whole experimental period, the gonad, gut, lantern indices and organic matter levels of different organs were determined at the end of the experiment. During the mature stage sea urchins in good nutritional conditions did not increase in size, but allotted energy to gonad production and stored reserves in body wall. Limiting food stopped the gonadal growth without complete regression. During the recovery period food allowed somatic growth, i.e. test growth and the storage of reserves in gonad somatic cells. This somatic production did not occur under food-limited conditions and the resources allotted for survival and maintenance were taken from different body components.     

Growth of Corbicula fluminea (bivalvia) fed artificial and algal diets

Laboratory experiments were conducted to assess the growth of Corbicula fluminea Müller (1774) fed mixed diets of the green alga Ankistrodesmus and five artificial foods (nine-grain cereal, rice flour, rye bran, denatured brewer's yeast and Ralston Purina trout chow). Clams lost weight after 30 days on each artificial diet; however, weight losses were significantly less than those of control clams which were not fed (p < 0.05). In contrast, clam growth was positive in each treatment which included Ankistrodesmus. There was no significant difference in the tissue growth of clams fed on an Ankistrodesmus monoculture and Ankistrodesmus supplemented with each artificial diet (p > 0.2). Therefore, we conclude that all of the artificial diets evaluated were of nutritional value. However, the alga Ankistrodesmus appears to be a superior food item. In subsequent experiments, clams were fed on diets consisting of various combinations of six genera of green algae (Pedinomonas, Ankistrodesmus, Chlamydomonas, Chlorella, Scenedesmus, and Selenastrum). All trialgal combinations of Chlamydomonas, Chlorella, Scenedesmus, and Ankistrodesmus and a dialgal culture of Chlamydomonas and Chlorella resulted in strong positive tissue growth. No other dialgal or monoalgal culture resulted in similar growth. Cultures of Selenastrum were found to be toxic, and their filtrate was determined to inhibit Corbicula filtration.