Community dynamics of intertidal soft-bottom mussel beds over two decades

Macrozoobenthos communities in the North Sea showed pronounced changes over the past decade in relation to an increasing number of invasive species and climate change. We analysed data sets spanning 22 years on abundance, biomass and species composition of intertidal soft bottom mussel beds near the island of Sylt (German Bight) in the Northern Wadden Sea, based on surveys from 1983/1984, 1990, 1993 and from 1999 to 2005. Mussel bed area and blue mussel biomass decreased, and a change in the dominance structure in the associated community comparing 1984 to mid-1990s with the period from 1999 to 2005 was observed. Coverage of the mussel beds with the algae Fucus vesiculosus decreased since the end of the 1990s. Within the study period biomass and densities of the associated community increased significantly. Dominance structure changed mainly because of increasing abundances of associated epibenthic taxa. Apart from the Pacific oyster Crassostrea gigas all other alien species were already present in the area during the study period. Community changes already started before Pacific oysters became abundant. An attempt is made to evaluate effects on the observed changes of decreasing mussel biomass, ageing of mussel beds, decreasing fucoid coverage and increasing abundances of invader. All four factors are assumed to contribute to changing community structure of intertidal mussel beds.     

Introduced Pacific oysters (Crassostrea gigas) in the northern Wadden Sea: invasion accelerated by warm summers?

Among the increasing number of species introduced to coastal regions by man, only a few are able to establish themselves and spread in their new environments. We will show that the Pacific oyster (Crassostrea gigas) took 17 years before a large population of several million oysters became established on natural mussel beds in the vicinity of an oyster farm near the island of Sylt (northern Wadden Sea, eastern North Sea). The first oyster, which had dispersed as a larva and settled on a mussel bed, was discovered 5 years after oyster farming had commenced. Data on abundance and size-frequency distribution of oysters on intertidal mussel beds around the island indicate that recruitment was patchy and occurred only in 6 out of 18 years. Significant proportions of these cohorts survived for at least 5 years. The population slowly expanded its range from intertidal to subtidal locations as well as from Sylt north- and southwards along the coastline. Abundances of more than 300 oysters m^–2 on mussel beds were observed in 2003, only after two consecutive spatfalls in 2001 and 2002. Analyses of mean monthly water temperatures indicate that recruitment coincided with above-average temperatures in July and August when spawning and planktonic dispersal occurs. We conclude that the further invasion of C. gigas in the northern Wadden Sea will depend on high late-summer water temperatures.Communicated by H.D. Franke     

Documentation of sites of intertidal blue mussel (Mytilus edulis L.) beds of the Lower Saxonian Wadden Sea, southern North Sea (as of 2003) and the role of their structure for spatfall settlement

Field surveys (dating back to 1950) and aerial photograph series (dating back to 1966) were evaluated to determine sites of intertidal blue mussel (Mytilus edulis) beds at the Wadden Sea coast of Lower Saxony. Maps were prepared indicating sites of blue mussel beds during the last decades. A table gives additional information on the presence (or absence) of blue mussel beds at each site at the time of large-scale surveys. Altogether 187 sites of M. edulis beds were recorded in the investigation area. In spring 1996, there were still only 19 sites where mussel beds still occurred, although at 51 sites residual mussel-bed structures were present, e.g. shell bases of former beds or protruding patches (which had been occupied by M. edulis before the beds vanished) and open spaces. At that time, the majority of the sites contained neither mussel beds nor mussel-bed structures. The analysis of recent data confirmed that mussel larvae have preferred to settle in sites of present mussel beds and sites with bases of former mussel beds. There was no preferential selection of one of these categories (settled beds vs. shell bases). On the other hand, the presence of mussel beds or mussel bed structures is not obligatory for settlement, because sites without those structures were also re-settled by the spatfall in 1996, even though on a smaller scale.     

江苏海门蛎岈山牡蛎礁生态现状评价

基于2013—2014年间的生态调查结果,评价了江苏海门蛎岈山牡蛎礁的生态现状。无人机航拍结果显示,江苏海门蛎岈山分布有750个潮间带区牡蛎礁斑块,总面积约为201519.37 m2;与2003年相比,海门蛎岈山牡蛎礁面积约下降了38.8%。活体牡蛎的平均盖度约为66%,2013年5和9月熊本牡蛎Crassostrea sikamea的平均密度分别为(2199±363)个/m2和(2894±330)个/m2。2013年5月海门蛎岈山熊本牡蛎种群的平均肥满度(CI)和性腺指数(GI)分别为(9.76±0.95)%和(1013±82)mg/g,均显著低于浙江象山港养殖的熊本牡蛎种群(P0.05)。海门蛎岈山熊本牡蛎的单倍体多样性和核苷酸多样性指数分别为0.119和0.00028,均高于长江口野生种群和浙江象山港养殖种群。海门蛎岈山熊本牡蛎种群受到尼氏单孢子虫(Haplosporidium nelson)的轻度浸染,其感染率(17.2%)低于浙江象山港养殖群体(47.3%)。泥沙沉积和人类捕捞是江苏海门蛎岈山牡蛎礁面临的主要胁迫因子,今后牡蛎礁恢复的重点是增加附着底物的数量。     

Heterogeneity and tissue specificity of tropomyosin isoforms from four species of bivalves

Heterogeneity and tissue specificity of tropomyosin isoforms obtained from four species of bivalves (Scapharca broughtonii (ark shell), Mytilus galloprovincialis (mussel), Atrina pectinata (surf clam) and Crassostrea gigas (Pacific oyster)), were examined. Tropomyosins were extracted from translucent and opaque portions of posterior adductor muscle, respectively, and cardiac muscle of each bivalve. There were two tropomyosin isoforms in the ark shell, the surf clam and the Pacific oyster. They were designated as TMa and TMb. In the ark shell, TMa was the common isoform and TMb was specific for the opaque portion of the adductor muscle. In the surf clam, TMb was the common isoform present in all tissues. TMa was found only in the translucent portion of muscle. In the Pacific oyster, TMb was the major component in both portions of adductor muscle and TMa was the major component in cardiac muscle, although both tropomyosins were included in all tissues. The mussel had only one tropomyosin.     

Histopathology in Pacific oyster (Crassostrea gigas) spat caused by the dinoflagellate Prorocentrum rhathymum

The recognition of an apparent association between seasonal oyster spat mortalities (up to 40%) and high Prorocentrum rhathymum density in the Little Swanport Estuary, Tasmania, prompted further experimental investigation into the toxicity by this dinoflagellate. Standard brine shrimp, haemolysis assays and intraperitoneal mouse bioassays revealed fast acting toxins in methanol but not aqueous extracts of P. rhathymum, with mice dying in less than 20 min. Oyster bioassays involved feeding spat (4 mm shell width) for 21 consecutive days on a diet of cultured P. rhathymum at simulated bloom densities (10⁴ cells ml⁻¹). No oyster mortality was observed, however, histopathological signs of thin, dilated gut tubules and sloughing of gut cells resembled those seen in affected field samples. In contrast to field samples, gill pathology was also observed in experimental exposure oysters.     

Distribution and Seasonal Abundance of Trematode Parasites (Trematoda: Allocreadiidae: <Emphasis Type="Italic">Crepidostomum</Emphasis> spp.) in burrowing-mayfly nymphs (Ephemeroptera: Ephemeridae: <Emphasis Type="Italic">Hexagenia</Emphasis> spp.) from Connecting Rivers of the Laurentian Great Lakes

Burrowing-mayfly nymphs such as Hexagenia spp. have been used extensively in North America and Europe as a biomonitoring tool to indicate mesotrophic water quality, yet infestation by associated parasites has not been well documented. We performed laboratory analysis of archived samples of Hexagenia spp. nymphs collected in 1985 and 1986 to provide base-line data on the distribution (1985) and seasonal infestation (1986) of the trematode parasite Crepidostomum spp. in Hexagenia spp. nymphs in connecting rivers between Lakes Superior and Erie of the Laurentian Great Lakes. In May and June 1985, frequency of occurrence of metacercariae was widely distributed throughout the connecting rivers (63% of 203 stations with nymphs), except in areas where nymph densities were relatively low (i.e.,≤69 nymphs/m²). Distribution was probably underestimated in the present study because of low probability (mean = 31%, range = 0–57%) of detecting infestation in a small number of collected nymphs ( ≤10) at nymph densities ≤69/m². In 1986, seasonal infestation between April and October occurred in 3.3% (627) of 18696 nymphs. Overall prevalence, mean intensity, and mean abundance of parasites at one station in the St. Marys River indicate parasite transmission occurred between June and September. This period of transmission is dependent on the life-cycle of the parasite. In addition, the life-cycle of Hexagenia spp. determines which annual cohort of nymphs is infested and therefore, the duration of infestation. Although, no impacts of infestation on Hexagenia spp. nymphs were observed in the present study, infestation intensities were high enough (≥25 metacercariae per nymph) at one station in the St. Marys River to potentially cause tissue damage in a high proportion (53%) of infested nymphs.     

Specific Detection of Pacific Oyster (Crassostrea gigas) Larvae in Plankton Samples Using Nested Polymerase Chain Reaction

Management of sustainable Pacific oyster fisheries would be assisted by an early, rapid, and accurate means of detecting their planktonic larvae. Reported here is an approach, based on polymerase chain reaction (PCR), for the detection of Pacific oyster larvae in plankton samples. Species-specific primers were designed by comparing partial mitochondrial cytochrome oxidase subunit I (COI) sequences from Crassostrea gigas, with other members of the family Ostreidae including those of Crassostrea angulata. Assay specificity was empirically validated through screening DNA samples obtained from several species of oysters. The assay was specific as only C. gigas samples returned PCR-positive results. A nested PCR approach could consistently detect 5 or more D-hinge-stage larvae spiked into a background of about 146 mg of plankton. The assay does not require prior sorting of larvae. We conclude that the assay could be used to screen environmental and ballast water samples, although further specificity testing against local bivalve species is recommended in new locations.     

Immunohistochemical localization and radioenzymatic measurements of serotonin (5-hydroxytryptamine) in hearts of Aplysia and several bivalve mollusks

Summary Serotonin immunoreactivity was localized in hearts of the opisthobranch gastropod, Aplysia californica (sca hare) and several species of bivalve mollusks, the heterodonts, Mercenaria mercenaria (quahog or cherry stone clam), Protothaca staminea (little neck clam), and the pteriomorphs, Hinnites multirugosus (rock scallop), Crassostrea virginica (eastern oyster), Mytilus edulis (eastern mussel), and Geukensia demissa (ribbed mussel). In addition, serotonin was assayed in the ventricles, auricles and heart-associated tissues in A. californica, M. mercenaria, H. multirugosus, and G. demissa with a sensitive radioenzymatic assay. Serotonin concentrations and the density of innervation were significantly higher in members of the subclass Heterodonta compared to the subclass Pteriomorpha. Serotonin immunoreactivity was observed in all species surveyed except G. demissa, which also contained relatively low concentrations of serotonin. Varicose fibers presumably corresponding to release sites were localized in the ventricles, auricles, and the auricular-ventricular valves. We hypothesize that in the species where serotonin-immunoreactive fibers are present, serotonin serves to modulate cardiac myogenic activity. The significance of the observed distribution and concentration of serotonin to the physiological effects of serotonin on cardiac function in these species is discussed.     

Characterization of six microsatellite loci in the Baltic blue mussel <Emphasis Type="Italic">Mytilus trossulus</Emphasis> and cross-species amplification in North Sea <Emphasis Type="Italic">Mytilus edulis</Emphasis>

The blue mussel Mytilus trossulus occurs in the Pacific and in the North Atlantic. We developed and characterized six microsatellite loci for Baltic M. trossulus. Seventeen microsatellite loci were screened, of which six were polymorphic. The number of alleles among 50 individuals ranged from 3 to 13 and the observed and expected heterozygosity were 0.09–0.46 and 0.34–0.86, respectively. The loci were also tested for cross amplification in M. edulis, in which four of the six microsatellite loci were successfully amplified.     

Impact of the invasive parasitic copepod Mytilicola orientalis on native blue mussels Mytilus edulis in the western European Wadden Sea

Invasive species can indirectly affect native species by modifying parasite–host dynamics and disease occurrence. This scenario applies to European coastal waters where the invasive Pacific oyster (Magallana gigas) co-introduced the parasitic copepod Mytilicola orientalis that spills over to native blue mussels (Mytilus edulis) and other native bivalves. In this study, we investigated the impact of M. orientalis infections on blue mussels by conducting laboratory experiments using controlled infections with larval stages of the parasitic copepod. As the impact of infections is likely to depend on the mussels’ food availability, we also tested whether potential adverse effects of infection on mussels intensify under low food conditions. Blue mussels that were experimentally infected with juvenile M. orientalis had a significantly lower body condition (11–13%) compared with uninfected mussels after nine weeks of infection. However, naturally infected mussels from a mixed mussel and oyster bed did not significantly differ in body condition compared with uninfected mussels. Contrary to effects on mussel condition, we did not find an effect of experimental infections on clearance rates, shell growth or survival of blue mussels and no clear sign of exacerbating effects of food limitation. Our study illustrates that invasive species can indirectly affect native species via parasite co-introductions and parasite spillover. The results of this study call for the integration of such parasite-mediated indirect effects of invasions in impact assessments of invasive species.     

Perkinsus marinus in pleasure oyster Crassostrea corteziensis from Nayarit, Pacific coast of México

Culture of the pleasure oyster Crassostrea corteziensis is emerging as an alternative to the Pacific oyster (Crassostrea gigas) for oyster producers, who face severe mortalities since 1997 in Northwest México. For determining the health status of this species, we conducted a histopathological analysis of cultured populations from two estuaries in the Pacific coast of México. Macroscopical analysis revealed animals with transparent and retracted mantle. Histopathological analysis of these specimens showed tissue alterations and parasitic forms consistent with Perkinsus sp. infection. Stages of the parasite identified included tomont and trophozoites with an eccentric vacuole characteristic of Perkinsus spp. Pieces of tissues of infected oysters were incubated in Fluid Thioglycollate Medium (FTM) resulting in blue–black hypnospores after incubation. The identity of the parasite was confirmed by species specific PCR-based assay in DNA samples from oysters, tissue fractions from FTM cultures, and deparaffined samples with Perkinsus-like parasite detected by histology. Sequencing of positive amplified fragments (307 bp) showed a sequence similar to Perkinsus marinus strain TXsc NTS ribosomal RNA gene (100% coverage and 98% identity, GenBank Accession No. AF497479.1) and to P. marinus, Genomic DNA, (100% coverage and 97% identity, GenBank Accession No. S78416.1). The prevalence of P. marinus varied from 1 to 5% in Boca del Camichín and from 1 to 6% in Pozo Chino. In general, the intensity of infection was moderate. The infection was observed in oysters from 31 to 110 mm of shell length. This is the first record of P. marinus in oysters from the North America Pacific coast and the first record in C. corteziensis. The origin of this parasite in the area is unknown, but it may be associated to introductions of Crassostrea virginica from the East coast of United States of America or Gulf of México.     

A cDNA Microarray for Crassostrea virginica and C. gigas

The eastern oyster, Crassostrea virginica, and the Pacific oyster, C. gigas, are species of global economic significance as well as important components of estuarine ecosystems and models for genetic and environmental studies. To enhance the molecular tools available for oyster research, an international group of collaborators has constructed a 27,496-feature cDNA microarray containing 4460 sequences derived from C. virginica, 2320 from C. gigas, and 16 non-oyster DNAs serving as positive and negative controls. The performance of the array was assessed by gene expression profiling using gill and digestive gland RNA derived from both C. gigas and C. virginica, and digestive gland RNA from C. ariakensis. The utility of the microarray for detection of homologous genes by cross-hybridization between species was also assessed and the correlation between hybridization intensity and sequence homology for selected genes determined. The oyster cDNA microarray is publicly available to the research community on a cost-recovery basis.     

河北唐山曹妃甸-乐亭海域自然牡蛎礁分布及生态意义

牡蛎礁是生态系统服务价值最高的海洋生境之一,目前我国自然牡蛎礁分布和生态现状的基础信息仍然较缺乏。于2019年3月对河北唐山曹妃甸-乐亭海域自然牡蛎礁的空间分布、生态环境、牡蛎生物学和礁体动物群落开展了调查,并评估了该牡蛎礁的生态系统服务价值。该海域自然牡蛎礁分布于溯河(SR)、溯河口海域(SRE)和捞鱼尖海域(LYJ),总面积约15 km~2,是目前我国面积最大的自然牡蛎礁。基于96个牡蛎样品的16S rDNA检测,共识别出92个长牡蛎Crassostrea gigas、3个侏儒牡蛎Nanostrea fluctigera和1个巨蛎属未知种Crassostrea sp.。自然牡蛎礁中牡蛎平均密度介于104—3912个/m~2之间,不同礁区间牡蛎平均密度的大小排序为：SRE>SR>LYJ (P<0.05),平均生物量的大小排序为：SR>SRE>LYJ (P<0.05),平均壳高的大小排序为：SR>SRE=LYJ (P<0.05)。在该牡蛎礁内记录到49种礁体动物,其中软体动物16种、节肢动物16种、环节动物8种、棘皮动物5种、腔肠动物2种...     

Seasonal changes in energy allocation to somatic and reproductive body components of the common cold temperature sea urchin <Emphasis Type="Italic">Loxechinus albus</Emphasis> in a Sub-Antarctic environment

Monthly variations in the somatic indexes and energetic content of organs were investigated in Loxechinus albus from the Beagle Channel. Samples were collected monthly from May 2004 to May 2005. Lantern and test indexes did not vary significantly. A major peak of gonad index (GI) was observed in winter (sexual maturation period), with a strong declination in November suggesting a spawning period in spring. In coincidence a shortening of feeding activities was expressed by the lower values of gut index (Gut I), suggesting that the gut is a storage organ. The values of gonad energetic content (GEC) and total gonad energetic content (TGEC) showed minimum values in winter (ripe stage) and the maximum in spring (spawned stage). The TGEC reached higher monthly average values (50–200 kJ) than total gut energetic content (TGuEC) (20–40 kJ). These differences indicate that the gonads constitute the most important store organs in L. albus. Moreover, organic stores are built up in the gonads after spawning, and then utilized during gamete production.     

The Relationship between Disease in Crassostrea virginica (GMELIN) and Thermal Effluents in the Chesapeake-Delaware Bay Area

The null hypothesis for this research was that oyster parasites and disease were not associated with increased thermal loading in the Chesapeake-Delaware area. The American oyster Crassostrea virginica (GMELIN) was held at a reference station and a test station in the Chesapeake and Delaware Bay area for a year. Oysters from the four stations were sampled monthly for histological studies and parasite incidence together with the infestation of the polychaetous annelid Polydora websteri. The incidence of parasites and disease conditions from histology was negligible. Based on these results the null hypothesis was accepted, that oyster parasites and diseases did not increase with increased thermal loading. However, infestation of the oyster commensal P. websteri increased with increased temperature.     

Direct and indirect effects of Littorina littorea (L.) on barnacles growing on mussel beds in the Wadden Sea

On the extensive sedimentary tidal flats of the Wadden Sea, beds of the blue mussel Mytilus edulis represent the only major hard substratum and attachment surface for sessile organisms. On this substratum, the barnacle Semibalanus balanoides is the most frequent epibiont. In summer 1998, it occurred on over 90% of the large mussels (>45 mm shell length) and the dry weight of barnacles reached 65% of mussel dry weight. However, the extent of barnacle overgrowth is not constant and differs widely between years. Periwinkles (Littorina littorea) may reach densities >2000 m^–2 on intertidal mussel beds. Field experiments were conducted to test the effect of periwinkle grazing on barnacle densities. An experimental reduction of grazing and bulldozing pressure by periwinkles resulted in increased recruitment of barnacles, while barnacle numbers decreased with increasing snail density. The highest numbers of barnacles survived in the absence of L. littorea. However, a lack of periwinkle grazing activity also facilitated settlement of ephemeral algae which settled later in the year. Field experiments showed that the growth rate of barnacles decreased in the presence of these ephemeral algae. Thus, L. littorea may reduce initial barnacle settlement, but later may indirectly increase barnacle growth rate by reducing ephemeral algae. It is suggested that periwinkle density may be a key factor in the population dynamics of S. balanoides on intertidal mussel beds in the Wadden Sea.     

黄河口西南侧小岛河河口天然牡蛎礁的牡蛎种群结构

牡蛎礁是生态系统服务价值高、但退化最严重及受关注度最高的海洋生境之一,牡蛎礁修复亦成为国际海洋生态修复的热点。掌握牡蛎自然种群状况及动态变化是评估牡蛎礁修复效果的基础。目前,我国天然牡蛎礁的牡蛎种群状况相关的背景资料较为缺乏。在黄河口西南侧的小岛河河口新发现天然活体牡蛎礁,但该牡蛎礁曾被大规模的商业采捕,亟需推进针对性的保护和修复研究工作。基于2021年11月对该牡蛎礁开展的牡蛎种群生态调查,分析其种类组成、年龄结构及生长特征。结果显示:该牡蛎礁分布有近江牡蛎(Crassostrea ariakensis)和长牡蛎(Crassostrea gigas)。牡蛎礁上以活体牡蛎为主,死亡牡蛎壳体数仅占6.1%-6.7%。活体牡蛎的密度和生物量分别为(2811±778)个/m²和(21.97±30.43)kg/m²,近江牡蛎较多,其密度和生物量分别占比55.7%和76.4%。近江牡蛎和长牡蛎的年龄分别介于0+-4+ 龄和0+-2+ 龄,它们都以壳高介于30-40 mm及壳质量<5 g的0+ 龄个体数量居多(>80%)。近江牡蛎的壳体形态参数均值都高于同龄组长牡蛎的相应值。两种牡蛎壳体均呈负异速增长,不同龄级的壳体延展方向不同。拟合von Bertalanffy生长方程得到,近江牡蛎和长牡蛎的渐近壳高分别为286 mm和173 mm,估算的拐点年龄分别为5.47 龄和2.56 龄,两种牡蛎的生长曲线分布存在极显著差异(P<0.001)。以上结果表明,小岛河河口的天然牡蛎礁的牡蛎自然种群资源较丰富,具有高密度、低龄和低死亡率等特点,有较好的活力和扩张潜力,有利于被采捕后的礁体的恢复。两种牡蛎中,近江牡蛎因其具有较高的生物量和较长的生长年龄,对礁体形成和扩繁可能更为重要。建议对该天然牡蛎礁及牡蛎种群开展周期>3年的原位保护、修复和连续监测计划。     

Relationship between biofouling and growth of the pearl oyster pinctada fucata (Gould) in Kuwait,Arabian Gulf

Results are presented from a three-year investigation of the relationship between accumulations of marine fouling organisms and growth of the pearl oyster Pinctada fucata (Gould). Estimates are provided of the diversity of the foulers, and data on certain hydrological features of the experimental site are also given.There was an inverse correlation between growth of the pearl oyster and diversity of the biofouling assemblages, whatever depth. Growth curves did not follow seasonal variations in the environmental factors very exactly. It was also observed that the polychaete, Polydora vulgaris Mohammad, preferred the oysters planted on the bottom to those suspended near the surface. Bottom oysters suffered the highest mortality, but definite evidence of a relationship between mortality and infestation by the polychaete was not apparent. Percentages of both infestation and mortality were higher among the fouled oysters than those cleaned periodically.     

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.