TEP on the tide in killifish (<Emphasis Type="Italic">Fundulus heteroclitus</Emphasis>): effects of progressively changing salinity and prior acclimation to intermediate or cycling salinity

Transepithelial potentials (TEP) were measured in killifish, acclimated to freshwater (FW), seawater (SW), 33% SW or cycling salinities relevant to tidal cycles in an estuary, and subsequently subjected to salinity changes in progressive or random order. Random compared to progressive salinity changes in an upward or downward direction in FW- and SW-acclimated fish, respectively, did not greatly influence responses to salinity change. Fish acclimated to SW or 33% SW as well as those acclimated to cycling salinities behaved similarly (TEP more positive than +15 mV in 100% SW, decreasing to ~0 mV at 20–40% SW, and more negative than −30 mV in FW). In contrast, FW-acclimated fish displayed a less pronounced TEP response to salinity (0 mV in FW through 20% SW, increasing thereafter to values more positive than +10 mV at 100% SW). We conclude that when evaluated under estuarine tidal conditions, the killifish gill exhibits adaptive electrical characteristics, opposing Na⁺ loss at low salinity and favouring Na⁺ extrusion at high salinity, changes explained at least in part by the Cl⁻ to Na⁺ permeability ratio. Thus animals living in the estuaries can move to lower and higher salinities for short periods with little physiological disturbance, but this ability is lost after acclimation to FW.     

Influence of salinity on the migration of postlarval and juvenile flounder Pleuronectes flesus L. in a gradient experiment

The behaviour of the flounder Pleuronectes flesus was observed in an experimental aquarium that consisted of five connected units with decreasing salinity from 20 to 15, 10, 5 and 0·5. This experimental aquarium was highly effective for studying behaviour in salinity gradients, because the gradient was relatively stable. After 4 days of observation, significantly more flounder were found in water with a salinity of 0·5, indicating a preference for fresh water. The total length of the flounders ranged from 7·5 to 21 mm. The majority of larvae had already started eye‐migration, however, the longer specimens had completed metamorphosis. The migration towards low salinities was observed to increase with ontogenetic development.     

The growth of juvenile flounders (Platichthys flesus L.) at salinities of 0, 5, 15 and 35%%

Growth and food intake of juvenile flounders from the Kiel Bight were measured experimentally at different salinities (0, 5, 15 and 35 %). The growth at the middle levels (5 and 15 %) was faster then at 0 and 35 %. The reason for lower rates of increase in the freshwater group is the lower food intake. In the 35 % group the slower growth is caused by the worse food conversion. The variance of the individual growth rates in the 15 % salinity group remains far below the values of all other groups.     

Age/size effects on juvenile green sturgeon, Acipenser medirostris, oxygen consumption, growth, and osmoregulation in saline environments

The green sturgeon, Acipenser medirostris, is an anadromous species that migrates from freshwater (FW) to seawater (SW) relatively early in its life history, although the ages and sizes of juveniles at SW entry are not known. Developmental constraints of osmoregulatory organs may either prohibit (i.e., due to salinity tolerance limits) or minimize (i.e., due to substantial osmoregulatory or ionoregulatory energetic costs) SW entry in small fish. Interestingly, larger green sturgeon are often encountered in brackish water (BW) estuaries, perhaps due to an energetic advantage in occupying these near-isosmotic environments. To test hypotheses concerning fish-size effects on the energetic costs of occupying habitats of different salinities, we measured oxygen consumption rates in green sturgeon representing three age groups (100, 170, and 533 days post hatch; dph), which were acclimated for 5 weeks to one of three salinities (FW, <3‰; BW, 10‰; or SW, 33‰). Also, after 7 weeks, final wet masses were compared and blood and muscle tissue samples were taken to assess osmoregulatory abilities. There were no differences in body-mass-adjusted oxygen consumption rates between any salinities or ages, indicating that the energetic costs were not prohibitively high to occupy any of these salinities. The only mortalities occurred in the 100 dph SW group, where 23% of the fish died, from apparent starvation. Final wet masses were comparable between FW and BW for each age group and with the 533 dph SW group, but were lower in SW groups at 100 and 170 dph. Similarly, osmoregulatory abilities, in terms of plasma osmolality, Na⁺, K⁺, lactate, and protein concentrations, and muscle water content, were comparable in FW and BW groups at all ages, and with the SW group at 533 dph. These results indicated an age/body size effect in hyperosmotic adaptability, and that juvenile green sturgeon may be found in FW or BW at any age, but only have the ability to enter SW by 1.5 years (75 cm, 1.5 kg) of age.     

A critical analysis of transepithelial potential in intact killifish (Fundulus heteroclitus) subjected to acute and chronic changes in salinity

We investigated the in vivo salinity-dependent behavior of transepithelial potential (TEP) in Fundulus heteroclitus (3-9 g) using indwelling coelomic catheters, a technique which was validated against blood catheter measurements in a larger species (Opsanus beta; 35-70 g). In seawater (SW)-acclimated killifish, TEP was +23 mV (inside positive), but changed to -39 mV immediately after transfer to freshwater (FW). Acute transfer to dilute salinities produced a TEP profile, which rapidly attenuated as salinity increased (0, 2.5, 5 and 10% SW), with cross-over to positive values between 20 and 40% SW, and a linear increase thereafter (60, 80 and 100% SW). TEP response profiles were also recorded after acute transfer to comparable dilutions of 500 mmol L(-1) NaCl, NaNO3, Na gluconate, choline chloride, N-methyl-D-glutamate (NMDG) chloride, or 1,100 mosmol kg(-1) mannitol. These indicated high non-specific cation permeability and low non-specific anion permeability without influence of osmolality in SW-acclimated killifish. While there was a small electrogenic component in high salinity, a Na+ diffusion potential predominated at all salinities due to the low P Cl/P Na (0.23) of the gills. The very negative TEP in FW was attenuated in a linear fashion by log elevations in [Ca2+] such that P Cl/P Na increased to 0.73 at 10 mmol L(-1). SW levels of [K+] or [Mg2+] also increased the TEP, but none of these cations alone restored the positive TEP of SW-acclimated killifish. The very negative TEP in FW attenuated over the first 12 h of exposure and by 24-30 h reached +3 mV, representative of long-term FW-acclimated animals; this reflected a progressive increase in P Cl/P Na from 0.23 to 1.30, probably associated with closing of the paracellular shunt pathway. Thereafter, the TEP in FW-acclimated killifish was unresponsive to [Ca2+] (also to [K+], [Mg2+], or chloride salts of choline and NMDG), but became more positive at SW levels of [Na+]. Killifish live in a variable salinity environment and are incapable of gill Cl(-) uptake in FW. We conclude that the adaptive significance of the TEP patterns is that changeover to a very negative TEP in FW will immediately limit Na+ loss while not interfering with active Cl(-) uptake because there is none. Keeping the shunt permeability high for a few hours means that killifish can return to SW and instantaneously re-activate their NaCl excretion mechanism.     

Body water content over a range of ambient salinities in the sheepshead minnow

Body water content was determined on groups of Cyprinodon variegatus after sequential acclimation to ambient salinities in the range from fresh water to 100%. A multiple linear regression model applied to the data included body mass ( M ) of individuals and acclimation salinity ( S ) as independent variables, and body water content ( W ) as the dependent variable. The model that described the relationship was W (g) = 0·059 + 0·723 M (g)–0001 S (%). Converting values of body water content to percent of wet body mass produced a range of values from 74·5% in fresh water to 72·0% at 100%, only a 2·5% difference in body water content over this wide range of ambient salinities.     

Bracteoles affect germination and seedling establishment in a Mediterranean population of Atriplex portulacoides

The germination and subsequent seedling establishment of Atriplex portulacoides from a population in SW Spain were investigated in response to the presence or absence of attached bracteoles, at a range of NaCl concentrations (0–6%). Increasing salinity reduced both the final germination percentage and the speed of germination. The presence of bracteoles greatly inhibited germination and this effect was more marked with increasing salinity; bracteoles completely inhibited germination at salinities higher than 2%. The effects of salinity were substantially reversible, as cumulative germination after transfer of ungerminated seeds to distilled water was similar in all salinity treatments. This enforced dormancy could serve to prevent germination of floating fruits during dispersal in seawater. However, after transfer to distilled water the germination of seeds with bracteoles was still significantly lower than those without them, as a result of both higher dormancy and lower viability (as revealed by tetrazolium testing). Bracteoles thus also appear to enforce some physical dormancy. Seedlings derived from seeds with bracteoles demonstrated higher survival rates in fresh water than those derived from seeds without bracteoles but at 2% salinity there was no difference, and at higher salinities no seedlings of either type survived.     

Effects of salinity on growth and survival in five Artemia franciscana (Anostraca: Artemiidae) populations from Mexico Pacific Coast

Salinity is an important factor influencing growth and survival of aquatic organisms such as Artemia, a valuable aquaculture species. This study evaluated the effects of salinity on A. franciscana populations from different water bodies in Mexico's Pacific Coast. With this purpose, five autochthonous bisexual Artemia populations were tested to assess their survival and growth values against salinities of 40, 60, 80, 100 and 120 g/l, under laboratory conditions (25 +/- 2 degrees C; pH 8-10; constant light and aeration). The organisms were fed with 100 mL of rice bran and 2L of Tetraselmis suecica (500 000 cel/ml). The culture experiments were made in 200L plastic tanks, and survival and growth final values were obtained after 21 culture days. Survival and growth curves were determined by a regression analysis (R2). The significant differences between salinities were determined by ANOVA test (p < 0.05). The best survival and growth rates were found at salinities of 100-120 g/l. When the Mexican Artemia populations were cultivated at 40 g/l of salinity, 100% mortality was observed in the juvenile stage. This study determined that survival and growth values of A. franciscana populations increased with salinity. The five A. franciscana populations presented significant differences in their survival rate under various salinity regimes. The studied populations experienced high mortality at salinities under 60 g/l and over 200 g/l, and especially during the metanauplius stage. The present study confirms that growth rates in Mexican A. franciscana populations from Pacific Coast habitats are not inversely proportional to salinity. These A. franciscana populations should be cultured at 100-120 g/l of salinity to obtain better survival and growth rates. This data is useful to improve culture systems in aquaculture biomass production systems.     

Ontogenesis of osmotic regulation in the striped mullet, Mugil cephalus L.

The osmoregulatory capabilities of juvenile striped mullet, Mugil cephalus L ., of three size-groups (20–29, 30–39 and 40–9 mm s.l.) were compared in a series of six salinities ranging from fresh water to full sea water. The two smaller size-groups were able to tolerate instantaneous transfer from the brackish water in which captured, to all salinities but fresh water, while the 40–69 mm group were tolerant of instantaneous transfer to all experimental salinities. At high environmental salinities, the osmotic regulatory capabilities improved with growth in size to an apparently definitive condition in fish of 40–69 mm s.l. The smallest of these individuals were estimated to be about 7.5 months old. The osmotic regulatory capability in waters of low salinity had reached a definitive state with the development of tolerance to such salinities. The ontogenetic pattern for Mugil cephalus is virtually identical to that of the Atlantic salmon, Salmo salar .     

Salinity effects on oxygen consumption, gill Na+, K+-ATPase and ion regulation in juvenile coho salmon

The metabolic response of juvenile coho salmon Oncorhynchus kisutch to different salinities was examined, using whole-animal oxygen consumption rates and gill Na⁺, K⁺-ATPase activities as indicators of osmoregulatory energetics. Coho salmon smolts were acclimated to fresh water (FW), isosmotic salinity (ISO, 10‰) and sea water (SW, 28‰) and were sampled for up to 6 weeks for plasma levels of cortisol, glucose and ions (Na⁺, K⁺, Cl⁻), gill Na⁺, K⁺-ATPase activity and oxygen consumption rates. Following an initial adjustment period, plasma constituents in SW fish returned to near-FW values, indicating that the fish were acclimated to SW by day 21. Gill Na⁺, K⁺-ATPase activities on days 21 and 42 were lowest in ISO, higher in FW and highest in SW. This result is consistent with the idea that less energy would be required to maintain ion balance in an isosmotic environment, where the ionic gradients between extracellular fluid and water would be minimal. Oxygen consumption rates of swimming fish (1 body length s⁻¹), however, did not differ significantly between the three test salinities after 6 weeks. The results of this study suggest that the metabolic response of juvenile salmonids to changes in salinity is dependent on life-history stage (e.g. fry v . smolt), and that oxygen consumption rates do not necessarily reflect osmoregulatory costs.     

Unidirectional Na(+) and Ca (2+) fluxes in two euryhaline teleost fishes, Fundulus heteroclitus and Oncorhynchus mykiss, acutely submitted to a progressive salinity increase

Na⁺ and Ca²⁺ regulation were compared in two euryhaline species, killifish (normally estuarine-resident) and rainbow trout (normally freshwater-resident) during an incremental salinity increase. Whole-body unidirectional fluxes of Na⁺ and Ca²⁺, whole body Na⁺ and Ca²⁺, and plasma concentrations (trout only), were measured over 1-h periods throughout a total 6-h protocol of increasing salinity meant to simulate a natural tidal flow. Killifish exhibited significant increases in both Na⁺ influx and efflux rates, with efflux slightly lagging behind efflux up to 60% SW, but net Na⁺ balance was restored by the time killifish reached 100% SW. Whole body Na⁺ did not change, in agreement with the capacity of this species to tolerate daily salinity fluctuations in its natural habitat. In contrast, rainbow trout experienced a dramatic increase in Na⁺ influx (50-fold relative to FW values), but not Na⁺ efflux between 40 and 60% SW, resulting in a large net loading of Na⁺ at higher salinities (60–100% SW), and increases in plasma Na⁺ and whole body Na⁺ at 100% SW. Killifish were in negative Ca²⁺ balance at all salinities, whereas trout were in positive Ca²⁺ balance throughout. Ca²⁺ influx rate increased two- to threefold in killifish at 80 and 100% SW, but there were no concomitant changes in Ca²⁺ efflux. Ca²⁺ flux rates were affected to a larger degree in trout, with twofold increases in Ca²⁺ influx at 40% SW and sevenfold increases at 100% SW. Again, there was no change in Ca²⁺ efflux with salinity, so plasma Ca²⁺ concentration increased in 100% SW. As the killifish is regularly submitted to increased salinity in its natural environment, it is able to rapidly activate changes in unidirectional fluxes in order to ensure ionic homeostasis, in contrast to the trout.     

Survival and behavioral characteristics of amphidromous goby larvae of Sicyopterus japonicus (Tanaka, 1909) during their downstream migration

To understand the ecology and environmental tolerances of newly hatched larvae of the amphidromous fish Sicyopterus japonicus during their downstream migration, the salinity tolerance of eggs, 0-15 day old larvae, and adults, and the temperature tolerance, specific gravity and phototaxis of hatched larvae were examined. Tolerances of adults were measured as survival after a 24 h challenge in freshwater (FW), brackish water (1/3 SW) and seawater (SW). The survival rate of adult S. japonicus was 100% in FW and 1/3 SW, while none survived in SW. Hatching success of eggs (30 eggs each) was significantly higher in FW (mean: 73%) and 1/3 SW (73%) than in SW (19%). Tolerance of newly hatched larvae to salinity and temperature was investigated in different combinations of salinities (FW, 1/3 SW and SW) and temperatures (18, 23 and 28 °C). Larval survival was significantly different in each salinity and temperature. Survival rate was significantly higher in 1/3 SW than in FW and higher in SW than in FW at 23 °C and 28 °C. At the latter part of the experiment, there was no survival in FW and at 28 °C. Survival was higher in lower temperatures, but larval development did not occur in FW. Specific gravity of newly hatched larvae was 1.036 at 28 °C and 1.034 at 23 °C. When exposed to a light source on one side of an aquarium, larval distribution was not affected. Our results indicated larval S. japonicus are more adapted to brackish water and seawater than freshwater, while the adults and eggs are more adapted to freshwater and brackish water than seawater. This is consistent with their amphidromous life history with growth and spawning occurring in freshwater and the larval stage utilizing marine habitats.     

Salinity-related fitness differences amongst GPI genotypes in the mosquitofish Gambusia holbrooki (Poeciliidae: Teleostei)

Salinity-induced mortality was examined in the mosquitofish Gambusia holbrooki to determine if natural selection associated with the tidal inundation of lower reach populations could potentially maintain glucose-6-phosphate isomerase ( GPI -2; E.C. 5.3.1.9) allele frequency variation observed at field sites. In vitro experiments established if survival was independent of GPI -2 genotype, and/or standard body length (S.L.). Bioassay results at 17%₀ were consistent with lower survival of GPI -2*88/88 homozygote genotypes, GPI -2*100/100 genotypes in juvenile size classes, and greater survival of GPI -2*109/100 genotypes; 100% mortality was observed at salinities of 30%₀. In the early stages of the 30%₀ bioassay, juvenile fish carrying each GPI -2 allele suffered similar, high rates of mortality. Following 19 weeks exposure to 4–8%₀ NaCl the frequency of GPI -2*88 alleles decreased in adult G. holbrooki (> 17 mm SL). Salinity-induced mortality was found to be a selective influence capable of creating and maintaining GPI -2 allele frequency differences among G. holbrooki subpopulations, and/or between sex/size cohorts within subpopulations, but only at intermediate salinities. Whether saltwater inundation at field sites maintains inter-population diversity and produces local adaptation will be determined by the age structure of the population during periods of exposure, fluctuations in salinity as determined by the relationship between rainfall and tidal regime, and the level of immigration from non-affected sites.     

Salinity tolerance of young catfish, Clarias lazera (Burchell)

Experiments were conducted to determine the adaptability of young catfish, Claria lazera , to various saline concentrations. Young catfish, total length 52–88 mm, were subjected to abrupt and gradual changes from fresh water (salinity = 0.14‰) to various salinities. Ninetyfour per cent survival occurred when the fish were transferred from fresh water to 25% sea water (salinity = 9.5‰ No fish survived higher salinities even through gradual change.     

Effects of reduced salinity on survival, growth, reproductive success, and energetics of the euryhaline polychaete Capitella sp. I

Physiological adjustment to water of reduced salinity requires energy expenditure. In this study we sought to determine the fitness costs associated with such adjustment in the euryhaline polychaete Capitella sp. I, and the extent to which such costs could be explained by increased rates of energy expenditure. In a series of experiments conducted at 20 degrees C, salinity was reduced from 30 per thousand to either 25, 20, 15, 12, or 10 per thousand within 72 h after the larvae had been induced to metamorphose. Juveniles were reared on fine, organic-rich sediment. Over the next 15-30 days, we determined survival, growth, fecundity, and rates of respiration and feeding (via fecal pellet production). Larval salinity tolerance was also determined. Juvenile survival at salinities as low as 12-15 per thousand was comparable to that at 30 per thousand. The lower limit of salinity tolerance was 10-12 per thousand at 20 degrees C for both larvae and juveniles. Juveniles grew significantly more slowly at 12-15 per thousand in six of the seven experiments. Fecundity, however, was generally highest at intermediate salinities of 20-25 per thousand, and comparable at 30 and 15 per thousand. No individuals released embryos at 12 per thousand over the approximately 30-day observation periods in any of the three experiments in which the worms were reared at this low salinity. Reduced growth rates were not explained by differences in rates of respiration at different salinities: at reduced salinity, respiration rates were either statistically equivalent to (P>0.10) or significantly below (P<0.05) those recorded for animals maintained at 30 per thousand. Lower growth rates at lower salinities were best explained by reduced feeding rates. Further studies are required to determine whether digestive efficiency, growth hormone concentrations, or reproductive hormone concentrations are also altered by low salinity in this species.     

Activity and respiration in the anemone, Metridium senile (L.) exposed to salinity fluctuations

Activity and respiration in the anemone, Metridium senile (L.), were monitored under both constant and fluctuating salinity conditions. During constant exposure to 50% sea water it was found that the animals retracted the tentacles and that the rate of oxygen consumption decreased by ≈50%. The same response was elicited from animals in 100% sea water in a contracted state. Animals exposed to continually fluctuating salinities were found to retract the tentacles, contract the body wall, and produce amounts of mucus during periods of decreasing salinities. These reactions were reversed during exposure to increasing salinity. Oxygen consumption never ceased entirely in animals exposed to dilute sea water and it was found that during declining oxygen tension M. senile regulated its oxygen consumption until the environmental oxygen tension fell to ≈30% saturation.     

The distribution of 0-group flatfish in relation to abiotic factors on the tidal flats in the brackish Dollard (Ems Estuary, Wadden Sea)

The distribution of 0-group flatfish was investigated in 1992 in the Dollard (Ems–Dollard estuary, Wadden Sea). 0-Group plaice, flounder and sole were not evenly distributed over the sampled locations. The spatial distribution pattern of 0-group flatfish in the Dollard changed during the investigation period. In the first week of sole presence, when the mean length of sole was 24–30 mm, salinity correlated significantly with sole density. The distribution of juvenile sole larger than 40 mm total length was affected by the elevation of the location: 0-group sole was restricted to the sampled site with the lowest elevation. The distribution of 0-group plaice was related to sediment: no juvenile plaice were caught at locations with more than 10% mud fraction in the sediment. The distribution of 0-group flounder was also correlated with sediment. Later in the year, salinity correlated negatively with the distribution of 0-group flounder. The influence of sediment composition is probably indirect and linked to the abundance of preferred food items, such as Corophium volutator . Abiotic conditions were suitable to 0-group plaice, flounder and sole.     

Food deprivation alters osmoregulatory and metabolic responses to salinity acclimation in gilthead sea bream Sparus auratus

The influence of acclimation to different environmental salinities (low salinity water, LSW; seawater, SW; and hyper saline water, HSW) and feeding conditions (fed and food deprived) for 14 days was assessed on osmoregulation and energy metabolism of several tissues of gilthead sea bream Sparus auratus. Fish were randomly assigned to one of six treatments: fed fish in LSW, SW, and HSW, and food-deprived fish in LSW, SW, and HSW. After 14 days, plasma, liver, gills, kidney and brain were taken for the assessment of plasma osmolality, plasma cortisol, metabolites and the activity of several enzymes involved in energy metabolism. Food deprivation abolished or attenuated the increase in gill Na⁺,K⁺-ATPase activity observed in LSW- and HSW-acclimated fish, respectively. In addition, a linear relationship between renal Na⁺,K⁺-ATPase activity and environmental salinity was observed after food deprivation, but values decreased with respect to fed fish. Food-deprived fish acclimated to extreme salinities increased production of glucose through hepatic gluconeogenesis, and the glucose produced was apparently exported to other tissues and served to sustain plasma glucose levels. Salinity acclimation to extreme salinities enhanced activity of osmoregulatory organs, which is probably sustained by higher glucose use in fed fish but by increased use of other fuels, such as lactate and amino acids in food-deprived fish.     

Regulation of photosynthesis and oxygen consumption in a hypersaline cyanobacterial mat (Camargue, France) by irradiance, temperature and salinity

Short-term effects of irradiance (0-1560 micromol photons m(-2) s(-1)), temperature (10-25 degrees C), and salinity (40-160) on oxygenic photosynthesis and oxygen consumption in a hypersaline mat (Salin-de-Giraud, France) were investigated with microsensors under controlled laboratory conditions. Dark O(2) consumption rates were mainly regulated by the mass transfer limitations imposed by the diffusive boundary layer. Areal rates of net photosynthesis increased with irradiance and saturated at irradiances >400 micromol photons m(-2) s(-1). At low irradiances, oxygen consumption increased more strongly with temperature than photosynthesis, whereas the opposite was observed at saturating irradiances. Net photosynthesis vs. irradiance curves were almost unaffected by decreasing salinity (100 to 40), whereas increasing salinities (100 to 160) led to a decrease of net photosynthesis at each irradiance. Dark O(2) consumption rates, maximal gross and net photosynthesis at light saturation were relatively constant over a broad salinity range (60-100) and decreased at salinities above the in situ salinity of 100. Within the range of natural variation, temperature was more important than salinity in regulating photosynthesis and oxygen consumption. At higher salinities the inhibitory impact of salinity on these processes and therefore the importance of salinity as a regulating environmental parameter increased, indicating that in more hypersaline systems, salinity has a stronger limiting effect on microbial activity.     

Gradual transfer to sea water of rainbow trout: effects on liver carbohydrate metabolism

The levels of glycogen and glucose, as well as of the activities of several key enzymes of glycogenolysis, glycolysis, gluconeogenesis and the pentose phosphate shunt were assessed in livers of small (97 g) and large (166 g) rainbow trout Oncorhynchus mykiss during gradual transfer to sea water (salinities of 0, 9, 18 and 28%). Plasma glucose and protein levels were also evaluated. Liver carbohydrate metabolism changed during gradual adaptation of rainbow trout to sea water. Glucose increased with salinity in livers of both sizes of animals, as did glycogenolysis and gluconeogenesis, but this latter only in large animals, and glycolytic rates in small animals. The different responses in metabolic parameters between sizes of animals may reflect a higher stress of the small animals and/or a better adaptation of large animals to increased salinities, in a way similar to that previously suggested by other authors after evaluation of non-metabolic parameters. The changes observed at 28% can also be the result of reduced food consumption.