Trophic interactions between the toxic dinoflagellate Alexandrium catenella and Mytilus chilensis: feeding and digestive behaviour to long-term exposure.

Trophic interactions between Mytilus chilensis and the toxic dinoflagellate Alexandrium catenella were studied over a 21-day period to determine the absorption kinetics of biochemical components and the enzymatic activity of the digestive gland (i.e. amylase, cellulase complex, laminarinase and protease). For the duration of the experiment, logarithmic relationships were established for the assimilatory balance and carbohydrase activities (amylase, laminarinase and cellulase) of the mussels fed with a toxic diet. The study shows that M. chilensis was able to develop mechanisms which allow exploitation of the toxic microalgae as a food source, despite that its feeding and digestive processes were affected during the first days of contamination.

Dynamics of biochemical components, lipid classes and energy values on gonadal development of R. philippinarum associated with the temperature and ingestion rate.

This study evaluates the effect of temperature, coupled with ingestion rate, on the dynamics of biochemical components and lipid classes in R. philippinarum. The data are discussed with regard to sexual development and energy balance. Experimental protocol developed in the present study used two groups of the clam R. philippinarum: L (temperatures of 14 degrees C and 18 degrees C) and H (temperatures of 18 degrees C and 22 degrees C). The intra-group ingestion level was similar, although the ingestion level of the clams in the group H was 2.4 times higher than group L. We observed that R. philippinarum conditioned at 18 degrees C (18L) shows higher protein content, furthermore an important loss of organic weight was observed after 48 days. In such a situation, the clams use their own reserves (carbohydrates and glycogen) for sexual development while in situations without food stress (positive energy balance) and low temperature (14 degrees C) an accumulation of reserves is produced. Strikingly dissimilar behaviour in biochemical composition was observed for the 18H and 22H treatments, both with a positive energy balance. Despite similar protein content, the highest levels of carbohydrates were observed at the lower temperature (18 degrees C). Glycogen was also higher for the 18 degrees C treatment, although the differences were significant only in the males. Although the total lipids in R. philippinarum showed no significant differences in any treatment, they became apparent and related to sex when considering the individual lipid classes. There was no variation in lipid classes in the males between the 14L and 22H treatments despite the large disparity in the degree of sexual development. However, in the females significant differences in lipid classes (phospholipids, triglycerides) were observed. The results of this study show that a positive energy balance permits R. philippinarum gonadal development and accumulation of reserves both in low and high temperature conditions. In low temperature situations, gonadal development is slower and the energy reserves are accumulated in the form of carbohydrates. When the clams are conditioned at high temperatures, gonadal development is fast and complete, carbohydrates are consumed and lipids are accumulated.

Response of two species of clams, Ruditapes decussatus and Venerupis pullastra, to starvation: physiological and biochemical parameters.

Adult specimens of two species of clams, Ruditapes decussatus and Venerupis pullastra, were starved for 84 days to determine the effect of starvation on their metabolism. The energy cost of metabolism in starvation was assessed using the oxygen consumption rates (respiration rates) of the clams and assessing the type of fuel used to provide this energy by analyzing their biochemical composition. Respiration rates decreased in both species after 15 days of starvation and remained at basal levels throughout the rest of the experimental period. Both during the first period and also after the metabolism had slowed down, the amount of energy consumed in respiration was higher in V. pullastra than in R. decussatus. The energy needed to maintain vital functions in both species is obtained from catabolism of body components, with a reduction in dry mass of the specimens, and consequently their energy content, in both species. This reduction was greater in V. pullastra given that energy demand is higher in this species. In both species carbohydrates made the largest contributions to energy output, followed by lipids in males and proteins in females. However, the energy contribution of each biochemical component differs according to species: R. decussatus obtains its energy from the catabolism of carbohydrates and proteins in equal proportion, while V. pullastra obtains it from proteins and lipids. In both species, albeit to a greater extent in R. decussatus, we observed that female specimens conserved their lipids until the later stages of the period of starvation (day 70), after which they started to metabolise their lipid components more intensely. The interspecific differences are interpreted in relation to the different habitat occupied by the two species.

Lipid composition of Ruditapes philippinarum spat: effect of ration and diet quality.

This study investigates the influence of the lipid composition of microalgal and commercial flour diets on the lipid classes and fatty acids of Ruditapes philippinarum spat. Aspects of the nutritional role of the diets and the feeding ration are discussed with regard to previously published spat growth data. Our results demonstrated that clams showed the best growth rates when fed with diets that supplied a larger quantity of lipids, further characterised by a high content of phospholipids and triacylglycerols. We observed a significant correlation between the amount of triacylglycerols (r=0.929, p<0.05) and phospholipids (r=0.781, p<0.05) supplied and spat growth. In addition, R. phillipinarum spat reached the highest percentages of triacylglycerols (about 12%) and the lowest percentages of phospholipids (about 60%) and sterols (about 4%) with these growth-promoting diets. Spat fed with the other diets and/or rations showed lower growth rates and significantly lower quantities of triacylglycerols. In the present study, the alternative essentiality of 20:5n-3 and/or 22:6n-3 is confirmed. The diminishing supply of 22:6n-3 without an increase of 20:5n-3 has an effect on the growth of the spat. The dietary composition of fatty acids influenced the fatty acid profiles in bivalves. The results of our study would suggest that R. philippinarum is incapable of transforming 18:3n-3 to 20:5n-3 and then to 22:6n-3. Accordingly, although diets contained 18:3n-3, the spat reflected the low content of 20:5n-3 of the diets. With respect to 22:6n-3, given that this fatty acid is present in high quantities in Isochrysis, the spat content of this fatty acid was relative to its content in the diet. This study showed that clams with the lowest growth rates presented an increase in 20:4n-6. In the spat fed with the microalgal diets, 18:2n-6 scarcely reached 2% whereas in spat fed with wheat germ 18:2n-6 amounted to 18% of the total fatty acids. This fatty acid, by means of elongation, transforms to 20:2n-6, which also appeared in important quantities in the lipids of the spat fed totally or partially by wheat germ. We note that 20:2n-6 did not originate from the diet since it is absent in the microalgae and the flour. The desaturation of 20:2n-6 to 20:3n-6 has not been observed (low levels of 20:3n-6 in all cases) and therefore it can be assumed that the observed levels of 20:4n-6 were diet-related.

Enzymatic and feeding behaviour of Argopecten purpuratus under variation in salinity and food supply.

Argopecten purpuratus is a bivalve filter feeder inhabiting protected areas of the Peruvian coastline and northern and central Chile. The species is commercially valuable and the natural stocks are over-exploited in Chile, mainly in the central region. Expansion of aquaculture farms to the geographically protected coastline of southern Chile is a potential solution to this problem. So, the objective of the present study was to determine the acute feeding and digestive response of A. purpuratus in relation to short acclimation periods (1, 3 and 7 days) and exposure to variable salinity (24 per thousand, 27 per thousand and 30 per thousand) with two different diets. The feeding behaviour of A. purpuratus follows that described for other species of bivalves whereby the decrease in salinity produces a reduction in the clearance and ingestion rates. This behaviour is not affected by the diet type or the 7 day acclimation period. Lower values of absorption efficiency are associated with increasing food concentration. The response of the AE at 24 per thousand salinity and low organic content diet indicates that the interaction of low salinity and low organic content diet reduces the AE to values approaching zero. The enzymatic investment of A. purpuratus at low salinity (24 per thousand) is similar for both diets and more reduced than for 30 per thousand and 27 per thousand salinity. The results show that osmotic stress leads to a reduction of the enzymatic response capacity. The multiple regression analysis indicates that different responses are observed as a function of the enzymatic activity and the organ. A reduction of the carbohydrase activities in the digestive gland was observed to low salinity (24 per thousand), specifically amylase and celullase. The results of this study are ecophysiologically relevant and highlight a relationship between digestive enzymatic activity and salinity, and may explain the possible differences in the energetic balance of A. purpuratus in response to changes in salinity.

Fatty acid profiles of Mytilus galloprovincialis (Lmk) mussel of subtidal and rocky shore origin.

Fatty acid profiles of seeds of the mussel Mytilus galloprovincialis originating from two habitats (rocky shore and subtidal) were compared after transfer to the same habitat (subtidal). The objective was to study the initial levels of different fatty acids of metabolic importance and, furthermore, the variability of these fatty acids over the experimental period. The results show that of all fatty acids identified in both seed groups, the polyunsaturated fatty acids (PUFA) is the group with highest percentage. Within this group, the C20:5n-3 and C22:6n-3 fatty acids show the highest levels. Additionally, the mussels of subtidal origin presented higher initial levels than the rocky shore mussels with regard to fatty acids characterised by energetic-type functions, such as the C14:0, C16:0, and the C20:5n-3 fatty acids, among others. Fatty acids characterised by structural-type functions, e.g. C18:0, C22:6n-3 and non-methylene interrupted dienoic (NMID) with 20 and 22 carbons in rocky shore mussels presented higher levels than those of the subtidal mussels. However, it has not been ruled out that aside from influences relating to the functional aspects of the different fatty acids, the initial differences were also associated with the quantitative and qualitative differences of the available food in both habitats. Nevertheless, 22 days into the experiment (in the majority of cases) the initial differences disappear in the different fatty acids with metabolic importance. On the basis of these results, the influence that mussel origin could exercise on the variability of the fatty acid profiles of recognised metabolic importance is discussed.

Enzymatic digestive activity in Mytilus chilensis (Hupé 1854) in response to food regimes and past feeding history.

Digestive enzyme activities (amylase, cellulase, laminarinase and protease) were analysed in mussels (Mytilus chilensis) from intertidal and subtidal habitats in Yaldad Bay, Chiloé, Chile. In order to analyse the effects of the past-feeding history (origin) and new nutritional conditions (habitat) on these enzymatic activities, a cross-over transplant was carried out and the analysis performed after a 7-day acclimation period. Crystalline styles showed higher carbohydrase and lower protease activities than digestive glands, with the highest differences recorded for subtidal mussels. Cellulase is the enzyme with the highest activity in both the digestive gland and crystalline style in all the experimental conditions. Intertidal mussels transplanted to a subtidal habitat showed enzyme resources significantly higher than in their original habitat. In the inverse case, mussels transferred from an original subtidal habitat to an intertidal one, a significant decrease in carbohydrase and protease activities was observed. The "past feeding history' is involved in the specific and total carbohydrase and protease activities, with a highly significant effect on amylase and cellulase activities in both the crystalline style and the digestive gland. Laminarinase activity can be interpreted considering the habitat (trophic regime), either individually or interacting with mussels' origin, in relation with the feeding periods. The results establish that in M. chilensis, an investment in enzyme resources is one of the mechanisms employed to optimise the acclimated response in terms of energy gains when variations in the food regime occur.

Effect of microalgal diets and commercial wheatgerm flours on the lipid profile of Ruditapes decussatus spat.

The influence of both the lipid composition of microalgal diets and commercial flours on the lipid classes and fatty acids of Ruditapes decussatus spat was studied. These aspects of the nutritional value of the diets were discussed in relation to the growth of the spat. Four diets were tested; Diet A, composed of 100% of the daily food ration of microalgae; Diet B, composed of 100% of wheatgerm; Diet C, composed of 50% of microalgae and 50% of wheatgerm; and Diet D, composed of 25% of microalgae and 75% of wheatgerm. The microalgal cells present a higher lipid content than that for wheatgerm. Tahitian Isochrysis cells have phospholipids and triacylglycerols as majority lipids, whereas in the wheatgerm particles, the lipids more abundant are triacylglycerols. Fatty acid content was higher in the microalgal cells than in the wheatgerm particles. The n-3 fatty acids were the most abundant acids in the microalgae, whereas the n-6 fatty acids were in the wheatgerm. The n-3 PUFA were not detected in wheatgerm. Phospholipids were the main lipids present in the clam spat, followed by triacylglycerols. Other lipid classes, detected in significantly lower amounts, included free fatty acids, sterols, and sterol ester + waxes. The composition of fatty acids in the spat was influenced by the fatty acid composition of the diet. Highest spat growth rates were observed with those diets that present a higher phospholipid/triacylglycerol relation. A negative correlation in the relation n-6/n-3 vs. growth has also been observed, with better growth rates in diets with a lower ratio. If the fatty acid 20:5n-3 and 22:6n-3 considered "essential" for marine animals were not present in the diet, they were not present in the spat either. Desaturation and elongation capabilities of R. decussatus spat were also discussed.

Changes in lipids of whole and minced rayfish (Raja clavata) muscle during frozen storage.

The effects of the storage temperature (-18 degrees C and -40 degrees C) and the addition of butylhydroxytoluene (BHT) on the different classes of lipids (phospholipids, triacylglycerides, free fatty acids, sterols and sterol esters plus waxes) and the fatty acid composition of minced (8 and 12 mm) and whole rayfish wing muscle stored for 1 year in the frozen state were studied. The phospholipid content decreased significantly and the free fatty acid content increased significantly at both storage temperatures, but more pronounced, at -18 degrees C. Significant differences were found between phospholipid and free fatty acid contents of the minced and the whole samples, which again were more pronounced at -18 degrees C. A significant increase of the major fatty acids (22:6n-3 and 16:0) was observed after 1 year in the frozen state. Significant differences were also obtained between the samples stored at -18 degrees C and at -40 degrees C; the lower the storage temperature, the higher their content. The polyunsaturated fatty acid (PUFA) content increased significantly in all the samples. Significant differences were found between the samples stored at -18 degrees C and at -40 degrees C. The lower the temperature, the higher the PUFA content. Non-significant differences were observed between the 8-mm and the 12-mm minced samples. Non-significant differences were found between the samples stored in the presence and in the absence of BHT. Mincing hastened hydrolytic and oxidative processes, which were slowed down at the lower storage temperature. Nonetheless, non-significant differences were found between both particle sizes.