兼养小球藻在不同浓度Fe3+培养下的蛋白质组学研究

为研究小球藻(Chlorella vulgaris)对不同浓度Fe~(3+)的响应,测定了自养和乙酸兼养小球藻在不同浓度Fe~(3+)条件下的生长速率和油脂含量,比较分析了兼养小球藻在不同浓度Fe~(3+)下的蛋白质组表达差异。结果表明,兼养小球藻比自养小球藻生长速率快,积累的生物量多,在缺铁条件下中性脂含量高。蛋白质组分析显示:在缺铁条件下光合作用相关蛋白含量最低;在缺铁和高浓度铁条件下,热激蛋白、蛋白合成和糖代谢等过程相关蛋白表达都下调,表明缺铁和高浓度铁条件对小球藻的生长都是逆境条件;在缺铁条件下氨基酸代谢相关蛋白表达上调,这可能是由于NO_3~–同化下降,氨基酸合成减少,需要进行氮的回收利用。上述结果表明:在兼养条件下缺铁培养的小球藻可获得较高生物量和中性脂含量;而高浓度铁能促进小球藻生长,但不能提高中性脂含量,对藻细胞也有一定的胁迫效应。     

Physiological response to high temperature in the Tropical Eastern Pacific coral Pocillopora verrucosa

The ecological and economic values of coral reef communities have encouraged efforts to implement periodic visual survey programs to secure their conservation. To date, visual monitoring‐based approaches have detected bleaching events weeks or months after the initial onset. An evaluation of the stress response of coral colonies, as well as their ability to resist and recover from the stress events, may increase our understanding of the physiological processes underling the stress and/or acclimation responses. Coral bleaching events, caused primarily by abnormally high temperatures, are continuously affecting coral communities worldwide. To evaluate the stress and recovery responses of the reef‐building coral Pocillopora verrucosa, a laboratory study was conducted herein. Coral nubbins were exposed to high temperatures to induce bleaching, and their ability to resist and recover from bleaching was subsequently monitored after returning the corals to ambient temperature. Lipid and chlorophyll concentrations, as well as Symbiodinium spp. density, decreased in samples exposed to nearly 31°C, slowly recovering to near‐control levels upon return to non‐stressful conditions. The present data set allows for an assessment of the vulnerability and ability of this common species to resist sub‐optimal environmental conditions, particularly the thermal stress events that will occur more commonly in their habitats as global seawater temperatures continue to rise.     

Metallothionein-like proteins induced by cadmium stress in the scallop Mizuhopecten yessoensis

Organisms have evolved a cellular response called stress protein response that increases their tolerance in adverse environmental conditions. Well known stress proteins that bind essential and toxic metals are metallothionein (MT). The scallop Mizuhopecten yessoensis is the most interesting organism because it is able to accumulate toxic cadmium in its digestive gland. However, in the tissue of the digestive gland of Mizuhopecten yessoensis MT (metallothioneins) have not been found. Eastern scallops, Mizuhopecten yessoensis, were collected from two locations ?? one clean and one polluted site. The concentrations of cadmium (Cd), copper (Cu) and zinc (Zn) were measured in the digestive gland. There was a significant increase in Cd concentrations in this studied tissue. We found that in the presence of cadmium Mizuhopecten yessoensis can induce high molecular proteins. The results of experiments have shown that Cd-binding ligands have a number of properties similar to MT: acetone and temperature stability; the ability to bind some metals, including Cd, Cu and Zn. Protein chromatography (FPLC, Superosa 12) from the digestive gland of scallop M. yessoensis has shown that cadmium is associated with high molecular weight Cd-binding proteins (72 kDa and 43 kDa). The major cadmium-binding protein 72 kDa is glycoprotein. In experiments we have demonstrated that Cd-binding proteins can be induced when there is cadmium exposure. The results of this study strongly suggest that the far eastern scallop Mizuhopecten yessoensis has a unique and well-developed system for the detoxification of heavy metals and it allows for biochemical systems to be maintained in a relatively stable manner in the presence of heavy metals.     

Determinants of species susceptibility to oxidative stress: A comparison of channel catfish and brown bullhead

Xenobiotic-mediated productions of reactive oxygen species, via enzymemediated redox cycling, have been implicated in a variety of toxicological phenomena including lipid peroxidation, enzyme inactivation and oxidative DNA damage leading to cancer. A comparison was undertaken of two benthic freshwater fish species that appear to differ markedly in their susceptibility to chemical carcinogenesis—the channel catfish (Ictalurus punctatus) and the more cancer-prone brown bullhead (Ameriurus nebulosus)-in terms of basic biochemical characteristics related to oxidative stress. This has included analysis of microsomal redox cycling of model xenobiotics (e.g. menadione) as well as antioxidant and other detoxifying enzymes in hepatic tissue of the two species. In addition, endpoints of oxidative stress, such as altered glutathione status and oxidative DNA damage, were examined. These studies have revealed numerous qualitative and quantitative differences between the two species both in terms of basal enzyme activities and in species response to model prooxidants. For example, bullhead appear to possess a greater capacity for microsomal redox cycling of xenobiotics, but have glutathione-dependent defense systems less able to withstand oxidative challenge. These and other interspecific differences have allowed for an improved understanding of the basic mechanisms which may underly species susceptibility to oxidative stress and critical manifestations such as cancer.     

Does thermal history influence the tolerance of temperate gorgonians to future warming?

To date, several studies have provided evidence that thermal stress affects the growth, survival and physiology of tropical and temperate macroinvertebrate species. However, few studies have focused on subtidal temperate species and the potential differential thermal tolerances of populations dwelling under contrasting temperature conditions. To assess the role that environmental history has on the response of the temperate gorgonian Eunicella singularis to thermal stress, we compared populations dwelling in the coldest and warmest areas of the NW Mediterranean Sea. Our results show that E. singularis populations from both areas exhibited a high resistance to thermal stress; however, populations from warmer areas had an increased tolerance to thermal stress. Specifically, the upper thermal limits found for cold and warm populations were 28 and 29 °C, respectively. The higher resistance of E. singularis colonies to thermal stress found in this study compared to the field temperature conditions during recent mass mortality events highlights that performing further thermotolerance experiments under contrasting levels of feeding is necessary to fully assess the tolerance thresholds displayed by both study populations. To our knowledge, this study provides the first evidence for the role of thermal history in shaping the thermotolerance responses of Mediterranean marine invertebrates dwelling under contrasting temperature environments.     

The influence of salinity on the heat-shock protein response of Potamocorbula amurensis (Bivalvia)

For biomarkers to be useful in assessing anthropogenic impacts in field studies involving aquatic organisms, they should not be affected by naturally occurring changes in environmental parameters such as salinity. This is especially important in estuarine environments and for relatively unspecific biomarkers like heat-shock proteins (hsps, stress proteins). In this study, the heat-shock protein response was measured in the euryhaline clam, Potamocorbula amurensis, after exposure to a range of salinities reflecting normal and extreme environmental conditions in Northern San Francisco Bay, California. The ability to raise cellular hsp70 levels in response to heat-shock was significantly impaired in P. amurensis collected from a low (0.5 ppt) salinity field site, and after 14 day exposure to low salinity in the laboratory.     

热锻炼对龙须菜高温胁迫响应的影响

夏季高温是限制龙须菜(Gracilariopsis lemaneiformis)生长和发育的一个主要问题。本研究以野生龙须菜作为材料,经过28℃的热锻炼预处理72 h,再分别置于32、36、40℃高温胁迫24 h,并测定了高温胁迫条件下龙须菜的丙二醛(MDA)、过氧化氢(H2O2)和可溶性蛋白质的含量,以及超氧化物歧化酶(SOD)活性和抗坏血酸(AsA)水平等生理指标,研究了龙须菜对高温胁迫的响应。结果表明:热锻炼能维持藻体较低的MDA含量,降低H2O2的产生速率,缓解藻体的膜脂过氧化伤害;同时热锻炼缓解了可溶性蛋白质的降解,增加了SOD酶的比活力,缓解了AsA的下降速率,说明热锻炼能缓解高温胁迫对藻体的伤害,有效地提高了龙须菜对高温胁迫响应的能力。     

A comparison of metal-binding proteins and cadmium metabolism in the marine molluscs Littorina littorea (gastropoda), Mytilus edulis and Macoma balthica (bivalvia)

Studies on the processes of Cd accumulation, turnover and storage in marine molluscs have revealed important qualitative and quantitative differences between species. In Littorina littorea collected from uncontaminated sites, Cd is predominantly bound to metallothionein (MT)-like proteins. Exposure to Cd, in the laboratory and field, results in an increase in Cd bound to these proteins, together with limited increases in Cd in the high molecular weight (HMW) pool (the latter partly a result of haemocyanin-mediated transport of Cd in the haemolymph). Polarographic determination of heat-stable MT-like compounds in L. littorea indicates there is little overall increase in the amount of protein produced in response to Cd exposure, suggesting that much of the newly incorporated metal may be bound to previously synthesised material. Relatively high levels of MT-like proteins, notably in the digestive gland (the major Cd-storage organ), appear to be an inherent feature of this species even in uncontaminated areas.MT-like proteins also sequester much of the Cd in exposed mussels Mytilus edulis, though, unlike Littorina, protein induction can be quantitatively related to exposure. In contrast, tellinid clams, Macoma balthica, fail to produce MT-like compounds, even in highly contaminated conditions, and Cd is mainly bound to HMW proteins.These variations in metal-binding behaviour probably explain the large interspecific differences in experimentally-determined Cd uptake rates and may account for differences in bioaccumulation potential observed in the field. Despite such variability, all three species studied appear to tolerate an influx of Cd provided the normal partitioning of metal is not irreversibly disturbed. Maintaining intracellular homeostasis, albeit at elevated Cd concentrations, probably compensates for the inability of these animals to regulate Cd uptake and for their extremely slow rates of Cd excretion.     

Structural integrity of Ellisolandia elongata reefs: A mechanical approach to compare tensile strengths in natural and controlled environments

Geniculate coralline algae are oases of biodiversity, providing nursery areas and shelter for the species that live amongst their fronds. The key to their success in the inter‐tidal is the ability to withstand hydrodynamic forces. Under culturing conditions most of the physical and ecological stressors such as intense hydrodynamic forces and grazing are extremely reduced, thus affecting species mechanical properties and their response to external threats. The aim of the present study was to investigate tensile mechanical properties of clusters of fronds of Ellisolandia elongata from natural (sheltered and exposed reef) and culturing conditions (after 1 month of culturing). The tensile test showed that the first failure stress (σ_I) was not significantly different between the natural and culturing conditions, indicating that the two reefs (sheltered and exposed) were characterized by the same distribution of pre‐existing, inherent structural flaws. Interestingly, the σ_max (maximum stress before rupture) was significantly different between the two conditions, with the culturing condition being more resistant to average load compared to the natural conditions. The maximum stress before rupture (σ_max) showed the influence of the environment in reducing the strength and elasticity of the fronds.     

Responses to Predation Cues and Food in Two Species of Sympatric, Tropical Sea Urchins

Abstract. We examined the responses of two tropical sea urchins, Lytechinus variegatus and Tripneustes ventricosus, to cues from predators, simulated predation events and food. Cues released from damaged conspecifics, heterospecifics and heteroclassics (holothurian) were used to simulate predation events in field experiments. Responses to the presence of seagrass, spiny lobster or both were tested in tanks. Findings were supplemented by natural history observations of dispersion patterns off Bermuda. In field experiments, neither species formed groupings in the presence of conspecific, heterospecific or heteroclassic cues. Flight responses were greatest in conspecific treatments; responses to heterospecific cues were intermediate to control and conspecific cues. Urchins in pre‐assembled associations remained in groups in control trials but dispersed when exposed to predation cues. Lytechinus exhibited greater sensitivity to predation cues than Tripneustes. Cues from a damaged sea cucumber invoked a response from Lytechinus but not Tripneustes. Both species employed a two phased response to cues from damaged conspecifics: initially a rapid, but ephemeral (2 min), alarm response followed by a slower (≈ 35 % lower) sustained flight phase for 6+ min, which in nature would disperse urchins downstream and away from a predator. In tank experiments, Lytechinus formed groupings only around food or food + predators. The presence of a predator reduced the aggregation response to food, suggesting that Lytechinus employed a risk aversion strategy. Tripneustes exhibited escape or refractory behavior in both control and experimental treatments in laboratory tanks.     

Depth influence on biochemical performance and thallus size of the red alga Gelidium corneum

Recent declines in the foundation species Gelidium corneum (Hudson) J.V. Lamouroux in pristine coastal stretches of the Southeastern Bay of Biscay have been documented in which individuals showed a high bleaching incidence. Among the potential factorsthat may be responsible for these changes in G. corneum abundance, it has been suggested that high levels of irradiance may be imposing stressful environmental conditions for shallow populations. Given that light is reduced exponentially as depth increases, in this investigation we examined the influence of depth on the stress of G. corneum by comparing the performance of several biochemical parameters and thallus size in plants collected at three different depths. The results revealed that plants growing in shallow waters showed lower antioxidant activity, lower concentrations of photosynthetic pigments (Chl‐a, total carotenoids, phycoerythrin), lower internal nitrogen levels (higher C:N ratio) and shorter thallus lengths than those living in deeper waters. Some of these results may be an expression of photoacclimation, but the low antioxidant activity and total carotenoid concentration detected in plants living in shallow waters suggest that the photoprotective mechanisms of G. corneum may have failed to offset photodamage. If so, our findings support previous studies that suggest that recent changes in solar radiation are partially responsible for G. corneum declines. Nevertheless, given that the metabolic performance of macroalgae is determined by the inter‐play of irradiance with other factors, including temperature and nutrient availability, further research is needed to reach a better understanding of the factors causing stress in G. corneum.     

Similar reproductive cycles and life-history traits in congeneric limid bivalves with different modes of nutrition

We have studied two congeneric limid bivalves, Acesta species novum and Acesta excavata (Bivalvia: Limidae), that live in similar physical conditions but use different food sources. Acesta sp. nov. live on the giant siboglinid tubeworm Lamellibrachia luymesi at cold seeps and feed on their eggs, a continuous food source that ultimately is derived from chemosynthesis, whereas A. excavata relies on planktonic food of photosynthetic origin, which varies seasonally. We hypothesized that these two species might show differences in the periodicity of reproduction; specifically, we predicted that A. sp. nov. would breed continuously and that A. excavata would demonstrate an annual reproductive cycle. Our hypothesis was refuted, as both species have a semi‐continuous reproductive cycle. Food quality, quantity or seasonality had no clear effect. Reproductive adults are functionally gonochoristic. Females broadcast buoyant eggs with an average size of 179 μm in A. sp. nov. and 160 μm in A. excavata. Both the size of the larval shell and the size of the spawned eggs are suggestive of pelagic lecithotrophic development. Both species also appear to be protandric hermaphrodites, changing from male to female at approximately 77 and 90 mm shell height, respectively, although not all individuals change sex. In the material investigated in this study there was a biased sex ratio favouring males.     

Correlation between the level of the potential biomarker, heat-shock protein, and the occurrence of DNA damage in the dab, Limanda limanda: a field study in the North Sea and the English Channel

In the present study, heat-shock protein of M(r) 70 kDa (HSP70), a marker of cellular stress response, was validated as a potential biomarker under field conditions. The dab, Limanda limanda (female, size > or = 25 cm, spawning maturity stage 2) was used as the indicator organism. The data on HSP level were correlated with the occurrence of DNA damage, measured in the same specimens of L. limanda, to prove the usefulness of the method. The area under investigation was the North Sea. Four locations were selected: station N01, close to Heligoland, in the North Sea; station N04 at the Dogger Bank; station N06 at the Firth of Forth; and station G08 in the English Channel. Ten animals from each location were selected and their livers used for the experiments. The results show that the highest levels of HSP70 (consisting of two forms of M(r) 75 and 73 kDa) were in fish from station N04, while low values were measured in livers from L. limanda collected at station N01. Intermediate levels were seen in the animals from the two other locations. By application of a novel technique, it was found that the extent of DNA damage (single-strand breaks and alkaline labile sites) in fish liver parallels the levels of both HSP70 forms. Our results suggest that L. limanda may be a useful bioindicator and heat-shock proteins, a useful biomarker for monitoring of environmental pollution.     

Comparison of ecophysiological characteristics between introduced and indigenous mangrove species in China

Due to its rapid growth, the introduced mangrove species Sonneratia apetala from Bangladesh has been widely used in mangrove restoration in southeastern China since 1985. As an indigenous mangrove species in Hainan, China, Sonneratia caseolaris was also planted in Guangdong Province for afforestation purposes. Both species have developed well in their new habitats, but their ecophysiological differences with the native mangrove species have not been studied. In this study, leaf gas exchange, water and nitrogen use efficiencies of two Sonneratia species were compared with those of selected native mangrove species (Avicennia marina, Aegiceras corniculatum, Kandelia candel, and Excoecaria agallocha) in Hainan and Shenzhen. The introduced S. apetala maintained lower carbon assimilation rate (A) and photosynthetic nitrogen use efficiency (PNUE) than the indigenous S. caseolaris. In Shenzhen, the two introduced Sonneratia had comparable photosynthetic rates and water use efficiency (WUE) with the native mangrove species, except that PNUE in S. caseolaris was significantly higher than in the native mangrove species. The two Sonneratia species showed significant overlap in PNUE and long-term WUE. Photosynthetic parameters derived from leaf photosynthetic light–response curves and A–C_i curves also suggested lower carbon assimilation capacities for the introduced Sonneratia than for the native mangrove species in both study sites. The lower light compensation point (LCP) of two introduced Sonneratia in both study sites also indicated a better adaptation to a low light regime than the native mangrove species. The results of photosynthetic capacities indicated that the introduced mangrove species have little competitive advantage over local native mangrove species in their respective new habitats.     

Induction of heat shock protein (hsp)60 in Isochrysis galbana exposed to sublethal preparations of dispersant and Prudhoe Bay crude oil

Adaptation to sublethal exposure to crude oil by phytoplankton is poorly understood. Use of chemical dispersants for oil spill remediation increases petroleum hydrocarbon concentrations in water, while exposing marine organisms to potentially toxic concentrations of dispersant. Heat shock proteins (hsps) have been found to serve as an adaptive and protective mechanism against environmental stresses. The objective of this project was to examine the induction of hsps in Isochrysis galbana, a golden-brown algae, following exposure to the water-accommodated fraction (WAF) of Prudhoe Bay crude oil (PBCO) and PBCO chemically dispersed with Corexit 9527® (dispersed oil: DO). Initial experiments using ³⁵S-labeled amino acids and 2-dimensional electrophoresis with subsequent western blotting identified and confirmed hsp60, a member of the chaperonin family of stress proteins, as being efficiently induced by heat shock in this species. One-dimensional SDS PAGE and western blotting, with hsp60 antibodies and chemiluminesence detection, were used to quantitate hsp60 following exposure to a range of environmental temperatures and concentrations of WAF and DO preparations. I. galbana cultured in 22 parts per thousand (‰) salinity showed a statistically significant increase (p<0.05) in hsp60 after exposure to 25, 30, and 35°C for 1 h compared to controls (20°C), while 34‰ cultures showed no significant increase in hsp60 with increasing temperature. Cultures (22 and 34‰) exposed to varying doses of WAF and DO exhibited statistically significant (p<0.05) increases in levels of hsp60 in all cases. A dose-related response was observed in all exposures (r²≥0.90) with the exception of 34‰ WAF. In independent exposures, a dose-related hsp60 response to naphthalene, a relatively abundant water-soluble aromatic hydrocarbon, was observed in both 22 and 34‰ cultures. Results of this study are consistent with previous studies in other species documenting increases in hsp60 levels with exposure to xenobiotics. Further studies are investigating the protective function of hsp60 against the toxic effects of exposure to WAF and DO preparations.     

Thermal sensitivity of native and invasive seabreams

This study compared the mortality and metabolic response of the Senegal seabream, Diplodus bellottii, an African species recently reported in Southern Europe and the white seabream Diplodus sargus, a native species, across a range of temperatures. The temperatures tested were 18, 26, 28 and 30 °C. Mortality was zero at 18 °C and very low at all other temperatures for both species, with the exception of D. bellottii, which experienced 32% mortality at 30 °C. Metabolic rates increased steadily with increasing temperatures, with a steep increase at 30 °C for D. bellottii. Thermal sensitivity ranged between 2 and 3 for both species and for all thermal intervals, with the exception of the thermal sensitivity between 28 and 30 °C for D. bellottii, which was 7. It was concluded that D. bellottii is under thermal stress at 30 °C. Diplodus bellottii may have expanded its distribution northwards due to an increase in sea surface temperatures. However, further warming may result in habitat loss for the juveniles, since Southern European estuarine systems will reach temperatures that may lead to lower fitness in juveniles of this species.     

Trophic relationships on a fucoid shore in south-western Iceland as revealed by stable isotope analyses, laboratory experiments, field observations and gut analyses

Rocky shores in the North Atlantic are known for their zonation patterns of both algae and animals, which can be expected to greatly affect food availability to consumers at different height levels on the shore. We tested the hypothesis that consumers would feed on the most abundant suitable food source in their surroundings. In total 36 species/taxa of common primary producers and consumers were sampled for stable isotope analyses from a sheltered fucoid shore at Hvassahraun in south-western Iceland. A selection of these species was also collected seasonally and from different height levels. Feeding experiments, field observations and gut analyses were also conducted. Our results were in good overall agreement with pre-existing knowledge of trophic relationships in the rocky intertidal. Consumers often appeared to be assimilating carbon and nitrogen from the most common diet in their immediate surroundings. The predator Nucella lapillus was thus feeding on different prey at different height levels in accordance with different densities of prey species. When tested in the laboratory, individuals taken from low on the shore would ignore the gastropod Littorina obtusata, uncommon at that height level, even when starved, while individuals from mid-shore readily ate the gastropod. This indicated that some kind of learned behaviour was involved. There were, however, important exceptions, most noteworthy the relatively small contribution to herbivores, both slow moving (the gastropod L. obtusata) and fast moving (the isopod Idotea granulosa and the amphipod Gammarus obtusatus) of the dominant alga at this site, Ascophyllum nodosum. The recent colonizer Fucus serratus seemed to be favoured. Selective feeding was indicated both by isotope signatures as well as by results of feeding experiments. Seasonal migrations of both slow and fast moving species could partly explain patterns observed.     

Properties of structure and function of cadmium-binding proteins/peptides from Euglena gracilis

Euglena gracilis makes two small Cd-binding proteins/peptides (Cd-BP I and Cd-BP II) in response to exposure to Cd²⁺. These proteins migrate slower than mammalian Cd-metallothionein (Cd-Mt) through Sephadex G-75, but like Cd-Mt also contain Zn and are separated into two species when chromatographed over a DEAE column. Both BPs are much less stable than Cd-Mt under acidic conditions. One source of this instability is that at least Cd-BP II contains acid-labile sulfide co-ordinated to the metals. It has been quantitated in Cd-BP II and shown to have a value of 1·25 ± 0·10 S²⁻/Cd. The reactivity of the Cd-binding sites in ligand substitution and thioldisulfide exchange reactions has been assessed. The reactions of Cd-BP I and II with pyridylazoresorcinol (PAR) were slow and displayed complex kinetic behavior. Similarly, both Cd-BPs react slowly and with complicated kinetics with 5,5′-dithiobis-(2-nitrobenzoate).     

A fish hepatocyte model for the investigation of the effects of environmental contaminants

An isolated fish hepatocyte culture system was developed as a models ystem to investigate the mechanisms of action of environmental contaminants. Hepatocytes were isolated from striped bass (Morone saxatilis) by an adaptation of the two stage perfusion technique of Seglen.¹ The system was used to evaluate metal binding protein (MBP) induction in response to cadmium, a primary inducer of metallothionein (MT) in rat hepatocytes. Striped bass hepatocytes appeared to be refractory to the induction of MBP by cadmium, since there was no significant increase in the synthesis of MBP for any of the doses at any of the time points investigated. However, when a similar experiment was performed using rat hepatocytes there was induction of MBP that was related to both dose and time. These comparative experiments indicate that although there are similarities between the hepatocytes of the two species in regard to ³⁵S incorporation into low molecular weight metal-binding proteins, there appear to be significant quantitative differences as well in regard to MBP kinetics. This in vitro model system could potentially be utilized to investigate the toxicological properties of other environmental contaminants.     

Influence of nutritional stress on digestive enzyme activities in juveniles of two marine clam species, Ruditapes decussatus and Venerupis pullastra

The potential use of digestive activities as indicators of the nutritional status in bivalves is discussed in relation to the results obtained in two clam species exposed to starvation and refeeding. Activities of some digestive enzymes (amylase, laminarinase, cellulase, and protease) were measured in juveniles of two commercially interesting species of clams, Ruditapes decussatus and Venerupis pullastra. The specimens were fed normally, being after subjected to a 15-days starvation and a further refeeding period. Samples were obtained at different moments of such feeding schedule to evaluate enzymes as well as weight (live, dry and organic) and length, in order to calculate growth rates and feeding efficiencies. Starvation led to a major decrease in clam growth as measured by dry weight and a negative growth as measured by organic weight, this coinciding with a certain degree of growth of the shell and a consumption of soft tissue. This response occurred more rapidly in R. decussatus but was of a lower magnitude than in V. pullastra. Activity of carbohydrases decreased rapidly in both species with starvation, although protease activity was maintained in R. decussatus. Recovery after the end of starvation was not similar in both species; while R. decussatus attained similar growth rates and enzyme activities to those measured prior to nutritional stress, V. pullastra only recovered 50% of its initial values. For both species of bivalves it can be concluded that digestive enzymes, and more specifically amylase, could be used as indicative of their nutritional condition.